2008 PROGRAM GUIDE Proficiency Testing Service AAB

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2 2008 PROGRAM GUIDE Proficiency Testing Service AAB AMERICAN ASSOCIATION OF BIOANALYSTS PROFICIENCY TESTING SERVICE 205 West Levee Street Brownsville, Texas (956) or (800) Fax (956) Web Site: BOARD OF SCIENTIFIC ADVISORS Everett S. Beneke, PhD, HCLD Microbiologist Safety Harbor, Florida John A. Boffa, HCLD Bioanalyst Howell, New Jersey Daniel H. Farkas, PhD HCLD Bioanalyst Baylor College of Medicine Houston, Texas Alvaro R. Garcia, MD Family Practice Houston, Texas Lynne S. Garcia, MS, MT(ASCP) Parasitologist Santa Monica, California Shawn Hartley, BLS Supervisor Baltimore, MD John H. Hughes, PhD Virologist Children's Hospital Columbus, Ohio J. Douglas Hutchison, PhD, CLD Bioanalyst North American Biologicals, Inc. Boca Raton, Florida Dennis W. Jay, PhD Clinical Chemist St Jude Children s Research Hospital Memphis, Tennessee Brooks A. Keel, PhD, HCLD Endocrinologist Reproductive Medicine Laboratories Wichita, Kansas Morton A. Levy, MD Hematologist/Oncologist Belleville, Illinois Pennell Painter, PhD, BCLD Bioanalyst University of Tennessee Memorial Hospital Knoxville, Tennessee Theodore J. Passon, Jr., PhD, BCLD Bioanalyst Pennsauken, New Jersey James E. Prier, PhD, HCLD Bioanalyst Blue Bell, Pennsylvania Patrick J. Quinn, PhD, HCLD Bioanalyst Carlsbad, California Richard G. Rawlins, PhD HCLD Bioanalyst Rush-Presbyterian St. Lukes Medical Center Chicago, Illinois Alvin M. Salton, HCLD Bioanalyst Community Medical Laboratory Metuchen, New Jersey Nicholas T. Serafy Jr., BA Director AAB Proficiency Testing Service Brownsville, Texas Warren Eric Vanderslice, PhD Technical Director AAB Proficiency Testing Service Brownsville, Texas AAB NATIONAL OFFICE 906 Olive Street, Suite 1200 St. Louis, Missouri Web Site: aab@aab.org Telephone (314) Fax (314)

3 AAB Proficiency Testing Service 2008 PROGRAM GUIDE GENERAL TABLE OF CONTENTS 2008 Schedule...2 General Instructions...3 Online Reporting Instructions...4 Reporting Form Instructions...5 Chemistry Clinical Microscopy & Urinalysis...9 Coagulation Embryology, Andrology & Fetal Fibronectin...10 Hematology Immunohematology...13 Immunology Microbiology Grading Procedures Table of Grading Limits Reinstatement Surveys...20 Reinstatement Surveys Order Form...21 General Attestation Form...23 Corrective Action Form...24 Corrective Checklist Form...27 GUIDE TO SPECIFIC PROGRAMS Acid-Fast Smears...15 Activated Clotting Time...9 Affirm Microbial Screen...15 Alcohol...6 Ammonia, Serum...6 Antinuclear Antibody...13 Antisperm Antibodies...10 Antistreptolysin-O...13 Bacteriology...15 Blood Gases...6 Blood Cell Identification (Hematology programs)...11 BNP...6 Cardiac Markers/Isoenzymes...6 Cardiac Markers/Isoenzymes, Plasma...6 C. difficile...16 Cell Identification...11 Centrifugal Hematology...11 Chemistry, Basic...6 Chemistry, Comprehensive...7 Chemistry, Urine...7 Chlamydia/GC/Strep Group B...16 Clinical Microscopy...9 CoaguChek PT, Basic...9 CoaguChek PT, Comprehensive...9 Coagulation, Plasma...10 C-Reactive Protein...13 Cryptosporidium/Giardia Immunoassay...16 D-Dimer...10 Direct Antiglobulin Testing...13 Direct Bilirubin...7 Erythrocyte Sedimentation Rate...11 Erythrocyte Sedimentation Rate-Rapid...11 Fecal Occult Blood...9 Fertility Endocrinology...7 Fetal Fibronectin (ffn)...10 Fructosamine...7 GC Culture...15 Glycohemoglobin...7 Gram Stain...17 Helicobacter Pylori Antibody...13 Hepatitis Markers...14 Hematology...11 Hematology w/diff A...11 Hematology w/diff B...12 Hematology w/diff C...12 Hematology w/diff D...12 Hematology w/diff E...12 Hematology w/diff F...12 Hematology w/diff G...12 HIV Markers...14 hscrp...14 Immunohematology, Basic...13 Immunohematology, Comprehensive...13 Immunoproteins...14 Infectious Mononucleosis...14 IVF Embryo Culture Media...10 Lipids...8 Lyme Disease...14 Oral Fluid HIV Antibodies...14 Parasitology...17 Pregnancy, Serum or Urine (HCG)...8 PTH/Insulin/C-Peptide...8 Reticulocyte Count...13 Rheumatoid Factor...14 Rubella...15 Special Chemistry...8 Sperm Count...10 Sperm Morphology...10 Sperm Viability...10 Strep Group A Antigen Screen...17 Syphilis Serology...15 Therapeutic Drug Monitoring...8 Throat Culture...15 Throat/Urine Culture...15 TIBC/UIBC...8 Tumor Markers...8 Urinalysis...9 Urine Culture...15 Urine Drug Screening...8 Urine Microalbumin/Creatinine...7 Viral Antigen Screen...17 Whole Blood Glucose...8 Whole Blood Prothrombin Time...10 AAB PROFICIENCY TESTING SERVICE ~ 2008 Program Guide 1

4 2008 PROGRAM GUIDE Proficiency Testing Service AAB 2008 SCHEDULE Program Guide ~ PROFICIENCY TESTING SERVICE AAB

5 AAB Proficiency Testing Service 2008 PROGRAM GUIDE GENERAL INSTRUCTIONS PRECAUTIONS All units of human blood or blood products used in preparing specimens, except those used in the Hepatitis and HIV Markers program, have tested negative for HBsAg, HCV antibody and HIV antibody at the donor level. Precautions described in CDC and FDA recommendations and OSHA blood borne pathogen rules should be followed when handling these specimens. These recommendations and rules include but are not limited to the following: 1. The specimens should be handled only while wearing impermeable gloves. 2. Gloves should be replaced if torn or contaminated. 3. Specimens should be opened in a hood or biological safety cabinet. 4. Do not pipet by mouth. 5. No eating, drinking or smoking is allowed in the laboratory. 6. Before leaving testing area, wash hands after removing gloves. 7. Specimens should be disposed of as hazardous waste. RECEIPT OF THE KIT 1. Samples are shipped via Federal Express. 2. notification of the FedEx tracking number is sent the day of each shipment to those participants that have registered for services. 3. Open immediately upon arrival. Check for damaged or missing specimens. 4. Call if replacements are required. Replacements will be sent only until the Monday following the shipment date. Hours are Monday thru Friday 8 a.m. - 5 p.m. Central Time. 5. Refrigerate specimens until they are analyzed. 6. Participants are responsible for notifying AAB when the testing kit has not been received within 3 days of the shipment date. 7. Alternate shipping dates cannot be provided. TESTING REGULATIONS The Clinical Laboratory Improvement Amendments of 1988 (CLIA '88) requires PT specimens to be analyzed as if they were patient specimens. To be in compliance you must not delay testing PT specimens unless your patient testing is also being delayed. Failure to participate in a testing event is unsatisfactory performance and results in a score of 0 for the testing event. Failure to return PT results or have them postmarked by the cut-off date displayed in the 2008 schedule on page 2 is unsatisfactory performance and results in a score of 0 for the testing event. REPORTING RESULTS 1. See the sample reporting form on page 5 for more specific instructions on executing the forms. 2. To submit results via the internet, please see the instructions for online reporting on page The attestation statement on the reverse of the Centers for Medicare and Medicaid Services (CMS)-scored analytes forms must be signed for each analyte by the analyst performing the procedure. In addition to the analysts' signatures, the director or the director's designee must sign only once for each reporting form. The designee for moderate complexity testing is the technical consultant while the technical supervisor is the designee for high complexity testing. There is no attestation statement for non-cms scored modules (1 & 2 vial programs). If your state or other regulating agency requires attestations for these programs, please use a copy of the general attestation form found on page Retain photocopies of your results and submit the red printed originals in the envelope provided. For online results, click the finish button to submit your results. Be sure to print the final results page which includes your confirmation number. No corrections can be made for any results without a confirmation number. 5. Telefaxes and photocopies are not acceptable. Originals are required for optical scanning. 6. Results must be postmarked by the cut-off date displayed in the 2008 schedule and on each reporting form. 7. AAB is not responsible for reports lost or delayed in the mail service. Late results will not be processed. 8. AAB is not responsible for results lost in the mail. Please consider taking advantage of our online reporting. If online reporting is not an option for you, consider the use of either an overnight service or a 2-day service that can provide confirmation of delivery. CODING YOUR METHODS A Reporting Form Instruction Sheet is included with each shipment. The back of this sheet lists method data on file only for analytes for which results were submitted in the most recent survey. Since this data will determine how currently reported results will be evaluated, please verify the entries in this list. If an entry is correct, do not recode it. If entries are missing or incorrect, please amend this list by coding/recoding method data directly on your results form(s). Methods for whole blood glucose and antigen screens must be entered each time. Do not return the Instruction Sheet. To code methods, ensure that you use current method codes by referring only to those listed in red and white reporting form, the General Instructions pages or the Method Code Addendum for the current event. Only method codes (not method descriptions) can be used to code participants' methods on the reporting forms. If no acceptable method description can be found in the method list, leave the method code blank, paper clip a package insert and/or method description to the report and we will code your method. Please write your AAB account number on the insert to allow us to match the new method with your results. If you do not report, or if you are late in reporting your results for an event, your method codes will be deleted from our database. You must recode all methods, reagents and instruments the following event or your results will be graded as Method Code Not Reported. Coding a method other than the one actually being used usually jeopardizes a participant's grading, even when results are transformed to mimic those of the method coded. We strongly recommend that participants report their results without transforming them to look like results from a method other than the one actually being used. Reporting untransformed results is not in violation of CLIA '88. CLIA '88 mandates that sample processing (not result processing) conform to that routinely applied to patient specimens. CMS SCORING INSTRUCTIONS To have your scores sent to CMS, your State Agency or COLA, place an "X" in the YES boxes on the reporting form. These boxes must be marked each reporting event. Scores will appear on the CMS cumulative score sheet only for those analytes that are checked YES. If the boxes are checked NO, no scores will be sent to CMS and will be indicated by a DC on the cumulative report unless state regulations override your request. Choose only one analyte within the following groups to report to CMS. If multiple analytes are marked, only scores from the first listed analyte will be sent to CMS. Antigen Screen Strep A Antigen Screen Chlamydia Antigen Screen GC Antigen Screen Strep B Antigen Screen C. Difficile Antigen Screen Chloride/Potassium/Sodium Basic Chemistry Blood Gases CK-MB Cardiac Markers/Isoenzymes Cardiac Markers/Isoenzymes, Plasma Coagulation Coaguchek PT, Comp Prothrombin Time (PT) Whole Blood Prothrombin Time Glucose Serum Glucose Whole Blood Glucose Hematology Cell Identification WBC Differential HCG/Pregnancy hcg/b-hcg (quantitative) Serum pregnancy, (qual-basic Ch) Serum pregnancy, (qualitative) Urine pregnancy (qualitative) HIV Anti-HIV-1 or 1/2 Screen Anti-HIV-1 Confirmation Anti-HIV-2 Screen Anti-HIV-2 Confirmation HIV-1 p24 Antigen Oral Fluid HIV Screen Oral Fluid HIV Western Blot Microbiology Bacteriology Gardnerella Affirm GC Culture Throat Culture Throat/Urine Culture Urine Culture Parasitology Cryptosporidium Giardia Parasitology Syphilis Serology FTA-ABS MHA-TP/TP-PA VDRL/USR EIA RPR, RST, trust Please follow all directions in filling out your reporting form completely, including the YES and NO boxes for CMS reporting. Once a report is graded, CMS does not allow for corrections due to participant s clerical and/or omission errors. CORRECTIVE ACTION If the evaluation of your results indicates that corrective action is necessary, the Corrective Action Form on page 25 and the Corrective Checklist Form on page 27 may be copied and completed for your internal records. Do not submit this form to AAB as it is for your internal use only. ADDRESS CHANGES Address changes must be received in writing by either fax or mail on institutional letterhead. Notice must be received 14 days prior to the next shipment date for the address to be updated. Participants will be responsible for shipping charges to resend a shipment due to a late address change. CANCELLATION POLICY A cancellation notice, on institutional letterhead, must be received by either fax, or mail 21 days prior to the shipping date in order to receive a credit or refund on a future shipment. Late enrollments received within the 21 days prior to the shipment waive the cancellation policy and no credit will be issued. The registration fee is nonrefundable. AAB PROFICIENCY TESTING SERVICE ~ 2008 Program Guide 3

6 2008 PROGRAM GUIDE Proficiency Testing Service AAB ONLINE REPORTING INSTRUCTIONS Go to and log on as soon as your samples arrive to verify your password and to avoid any last minute problems. Please refer to the reporting forms you received with your specimens for result codes, method codes and special instructions. If you prefer to mail your results, the envelope must be postmarked by the cut-off date located on your reporting form. We are not responsible for results delayed or lost that are sent by mail. Do not mail your results if you submit them online. You are required to sign and keep the Attestation that appears on the back of your reporting form for your records. If you have any questions pertaining to the submission of results via the internet please call (800) or (956) and ask for online support. For questions pertaining to methods, testing procedures, or specimen problems ask for technical support. After logging in please check the addresses we have on file by clicking edit profile. 1. Go to the Login page ( a. Enter your AAB Account Number & Password b. If this is your first time logging on you will find your initial password at the top of your reporting form. c. Click the radio button that pertains to the current shipment then click the Login Button'. 2. Change your password (if you have logged in before please skip to instruction line 4). a. From here you must enter your own password. b. You should also enter an address. c. Click Change Password. 3. You will receive an auto response after your initial login. 4. After you complete each form, click the finished button. Your form will be redisplayed along with your confirmation number. Please print that page for your records. Results are not accepted without a valid confirmation number. If you don t see the confirmation number on the form you printed please call immediately to verify that we received your report. 5. Methods codes do not have to be entered if we have your codes on file from the previous reporting period. If you did not submit results for the previous shipment then you will have to reenter your method codes. Please see the Method Data Sheet for the codes we have on file for your lab. If your method is not listed on the reporting forms please call technical support for instructions on reporting new methods. 6. After clicking the finished button your form will be locked. If you need to make changes to your form call (800) and ask for on-line support. 7. Please enter the less than (<) and greater than (>) signs when necessary in the result field along with your lowest or highest reportable range. When reporting on paper you must still report the < or > on a photocopy of your reporting form for clarity. 8. For qualitative results do not enter anything in the result fields other than the numeric codes listed on the forms. Characters such as + - n p pos neg will not be recognized and results will not be processed. 9. Once the cut-off date and time has passed you will no longer be able to login but you will still be able to view other areas of the website. Results reported on-line are downloaded at 5:00pm Central time on the posted date, therefore If you happen to still be logged on after the cut-off time, results may not be evaluated. 10. Graded results are not available on the website. You have the option of receiving graded results via by logging in and clicking edit profile and entering the addresses in the appropriate fields. Graded results are sent as soon as grading is finalized and are ed only once per reporting cycle as scheduled. A hard copy will follow by U.S. mail Program Guide ~ PROFICIENCY TESTING SERVICE AAB

7 AAB Proficiency Testing Service 2008 PROGRAM GUIDE REPORTING FORM INSTRUCTIONS These forms are designed to be read by a scanner which translates hand print into characters a computer can use thus reducing human intervention. Please exercise care in filling out these forms. To ensure that your results are translated correctly please follow the rules listed below. 1. Please adhere to the decimal points that appear on the form. Do not add decimal positions that do not appear on the form. This is also true if you report online using the web-based forms. The scanner will ignore any decimal points not already on the form. Example: If you enter a value such as "9.0" in a field that requires whole numbers, your result will be recorded as "90". It is also very important to zero fill all positions to the right of the decimal points where applicable. 2. Use a ball point pen with black or blue ink. 3. Place entries wholly within the red result boxes. Do not create or draw your own boxes as the red ink is a special ink that drops out. 4. Do not write comments on the form as the scanner will ignore non-numerical information and anything written outside of the reporting boxes. Please use a photocopy of your form for making comments or for reporting non-numerical information such as "less than (<)" or "greater than (>)" signs. 5. Do not draw lines across the page. If you do not perform a procedure, simply leave it blank. 6. Do not use highlighters. 7. Do not staple. 8. Do not punch holes. 9. Use White Out to make corrections. 10. Do not tape or stick anything to the form. 11. Submit the original red reporting form. Faxes or copies cannot be read by the scanner. 12. To have your scores sent to CMS (formerly HCFA), your State Agency or COLA, place an "X" in the YES boxes on the reporting form. 13. When calling, please have the AAB account number available so as to expedite your call. AAB PROFICIENCY TESTING SERVICE ~ 2008 Program Guide 5

8 2008 PROGRAM GUIDE Proficiency Testing Service AAB CHEMISTRY ALCOHOL Five liquid serum specimens per testing event. The specimens are for the analysis of ethanol. They contain 10 mg/dl of sodium fluoride as preservative. Code reagent from the list on the reporting form. Code instrument from the list found in the General Instructions or Addendum, which is included with each testing event. AMMONIA, SERUM Two liquid serum specimens per testing event. The specimens are for the analysis of ammonia. Code reagent from the list on the reporting form. Code instrument from the list found in the General Instructions or Addendum, which is included with each testing event. BLOOD GASES Five aqueous specimens per testing event for the following analytes: Chloride Potassium Ionized Calcium ph pco2 Sodium po2 Duplicate specimens are sent in each shipment. Participants must process these specimens in the following manner: (1) Before using, the specimens must be brought to room temperature (20-30ºC). If the material has been stored at temperatures above or below this range, remove ampules from the box and allow 4 hours for the material to equilibrate to room temperature. The gas-liquid equilibrium in each sealed ampule is dependent upon temperature. The most precise measurements will be obtained when storage temperature is controlled at 25ºC. The parameter most sensitive to temperature is po2; it changes inversely with temperature approximately one percent per degree. (2) To complete the equilibration of gas and liquid phases, shake the ampules 15 to 20 times (about 10 seconds) immediately prior to use. Hold the ampules at the tip and the bottom (with forefinger and thumb) to minimize temperature changes in the specimens. (3) Protect fingers with gauze, tissue or gloves and carefully snap off the neck of the ampule. (4) Introduce sample from the ampule within one minute after opening, using the procedure below which is appropriate to the transfer method required: a. Direct Aspiration: each specimen may be sampled directly from the ampule. b. Syringe Transfer: after opening an ampule, immediately insert a 20 gauge, 1 inch needle fitted to a clean, gastight syringe. Aspirate the specimen slowly from the bottom of the ampule without entrapping any air bubbles. Expel one or two drops through the needle, detach the needle from the syringe and immediately inject the sample directly into the instrument port. Code your instrument using the list at the bottom of the reporting form. Sodium, Potassium and Chloride Do not report results from vials labeled Basic Chemistry here. If also enrolled in the Basic Chemistry module, select only one set of Sodium, Potassium or Chloride results to be sent to CMS. Select the results from the Blood Gases module, only if you do not routinely test electrolytes on your main chemistry instrument. BNP Two lyophilized plasma samples per testing event for the analysis of BNP. Compatible with BNP and pro-bnp methods. Code reagents from the lists on the reporting form. Code instrument from the list found in the General Instructions or Addendum, which is included with each testing event. CARDIAC MARKERS/ISOENZYMES Five lyophilized specimens per testing event for the following analytes: CK2 (CK-MB), Qual Myoglobin CK2 (CK-MB), U/L, Troponin I ng/ml or % of Total Troponin T Reconstitute these specimens as follows: (1) Add 1.0 ml distilled or deionized water with a volumetric pipet. (2) Invert occasionally until completely dissolved (do not shake or vortex). (3) Let stand for 15 minutes after dissolution. (4) Invert again prior to sampling. Once reconstituted, the five Isoenzyme specimens should be processed as fresh patient serum. For laboratories performing confirmatory studies, additional testing material is available for a fraction of cost if needed. Code reagents from the lists on the reporting form. Code instruments from the list found in the Addendum, which comes with the specimens. CK2 (CK-MB) The reagent codes reported for CK2 (CK-MB) Isoenzymes reflect the units U/L, ng/ml or % of Total in which results are reported. However, participants must report their results directly, without conversion to other units. For example, participants using electrophoretic methods must report their results only in % of total enzyme, and participants using activity-based or mass-based methods must report their results only in U/L or ng/ml, respectively. Troponin Please test as soon as possible after receipt of specimens, within 5 days if possible. CARDIAC MARKERS/ISOENZYMES, PLASMA Five fresh frozen plasma specimens per testing event for the following analytes: CK2 / CK-MB, quant. Myoglobin Troponin I This survey is compatible only with the Biosite Triage. Code reagents from the lists on the reporting form. Code instruments from the list found in the Addendum, which comes with the specimens. CHEMISTRY, BASIC Five liquid serum specimens per testing event for the following analytes: Albumin Phosphorus Bicarbonate Potassium Bilirubin, Total Pregnancy (hcg), Serum Calcium Sodium Chloride Total Protein Cholesterol, Total Triglycerides Creatinine Urea Nitrogen Glucose Uric Acid Code instruments and reagents from the Reagent and Instrument Code lists, which come with the specimens. Calcium Results must be rounded to one decimal place and reported in mg/dl. To convert to mg/dl, multiply mmol/l by 4 and meq/l by 2. If reporting ionized calcium concentrations, code your reagent as 114 (all ionized calcium methods) for proper grading. Creatinine Vitros DT-60 participants using the 2-slide (ammoniabased) method should not report results based on diluted specimens if the diluted result is less than 7.5 mg/dl. These samples are not compatible with the Nova CRT enzymatic method. Electrolytes Diluted or indirect ISE methods are those in which the sample is introduced to the electrode along with the diluent. In undiluted or direct ISE methods, the sample is passed directly to the electrode without being diluted. Undiluted methods tend to give higher readings, especially for sodium and chloride. Because of this, correction factors may be employed to make undiluted methods agree with diluted ISE and flame photometer methods Program Guide ~ PROFICIENCY TESTING SERVICE AAB

9 AAB Proficiency Testing Service 2008 PROGRAM GUIDE CHEMISTRY Use reagent codes labeled "diluted ISE" or "dil ISE" if you use a diluted method. Use reagent codes labeled "und ISE" for ISE methods with no diluent and use reagent codes labeled "und ISE/flame eq." for undiluted ISE methods with a flame photometer correction factor. Glucose Do not report glucose values on the Basic Chemistry form using visual strip tests or instruments (usually hand-held) that generate whole blood glucose values. Whole blood glucose values are not equivalent to serum glucose values. Many whole blood instruments, such as the Abbott Vision and the Roche Reflotron do give serum glucose results and should be reported on the Basic Chemistry form. True whole blood glucose results can only be reported on specimens in the Whole Blood Glucose program. Pregnancy (hcg), Serum Basic and Comprehensive Chemistry vials can only be used report serum pregnancy results. Results obtained using Basic and Comprehensive Chemistry vials must be reported on the Basic Chemistry reporting form. If you are also enrolled in the Pregnancy module, you must report the results obtained from the Pregnancy vials on the Pregnancy reporting form. On the Pregnancy module, you may report either urine or serum results. You must select one set of results from among the serum pregnancy, urine pregnancy or quantitative hcg for CMS/COLA scoring. All other results will be graded, but the scores will not be reported to CMS/COLA as per CMS guidelines. CHEMISTRY, COMPREHENSIVE Five liquid serum specimens per testing event for the following analytes: Alanine Aminotransferase (ALT or SGPT) Alkaline Phosphatase Amylase Aspartate Aminotransferase (AST or SGOT) Cortisol Creatine Kinase (CK or CPK) Gamma Glutamyltransferase (GT or GGT) Human Chorionic Gonadotropin (beta or intact ) Iron Lactate Dehydrogenase (LD or LDH) Lipase Magnesium Thyroid Stimulating Hormone (TSH) Thyroxine, Free (FT4) Thyroxine, Total (TT4) Triiodothyronine, Total (T3) TUptake Code instruments and reagents (which include method principles) from the Reagent and Instrument Code lists, which come with the specimens. Lipase Report results in U/L. Magnesium Report this analyte s results on the proper line (according to the units used - mg/dl or meq/l). Thyroxine, Free (FT4) Report results in ng/dl. To convert pmol/l to ng/dl multiply by Triiodothyronine, Total (T3) Report results only in ng/ml. Divide ng/dl results by 100 to convert to ng/ml. TUptake Participants using the Abbott AxSYM, IMx or TDx; the Roche fluorescence polarization method or the Roche Elecsys should report results on the TUp Units/TBI line. The FT4 and Tuptake analytes are not compatible with the J & J Vitros/Eci instrument. CHEMISTRY, URINE Two liquid urine specimens per testing event for the following analytes: Amylase Phosphorus Calcium Potassium Chloride Sodium Creatinine Total Protein Glucose Urea Nitrogen Osmolality Uric Acid Only results from vials labeled "Chemistry, Urine" may be reported on the Chemistry, Urine form. Do not report results from the Basic & Comprehensive Chemistry vials on this form. Code instruments and reagents (which include method principles) from the lists found in the Reagent and Instrument Code list, which come with the specimens. URINE MICROALBUMIN/CREATININE Two liquid urine specimens per testing event. Code reagents from the lists on the reporting form. Code instrument from the list found in the General Instructions or Addendum, which is included with each testing event. DIRECT BILIRUBIN Two lyophilized specimens per testing event. Reconstitute these specimens as follows: (1) Add 1.0 ml reagent grade deionized water with a volumetric pipet. (2) Invert occasionally until completely dissolved (do not shake or vortex). (3) Let stand for 15 minutes after dissolution. (4) Invert again prior to sampling. Once reconstituted, the specimens should be processed as fresh patient serum. Code reagents from the lists on the reporting form. Code instrument from the list found in the General Instructions or Addendum, which is included with each testing event. FERTILITY ENDOCRINOLOGY Two liquid serum specimens per testing event for the following analytes: DHEA-s Estradiol Follicle-Stimulating Luteinizing Hormone (LH) Hormone (FSH) Progesterone Code reagents from the list on the reporting form. Code instrument from the list found in the General Instructions or Addendum, which is included with each testing event. FRUCTOSAMINE Two liquid serum specimens per testing event. Code reagents from the list on the reporting form. Code instrument from the list found in the General Instructions or Addendum, which is included with each testing event. Report on the appropriate line of the reporting form, depending on whether your method is calibrated in µmol/l of glycated polylysine (plys) or in mmol/l of 1-deoxy-1-morpholino-fructose (DMF). Samples are not compatible with the LXN Duet. GLYCOHEMOGLOBIN Two specimens per testing event. Code reagents from the list on the reporting form. Code instrument from the list found in the General Instructions or Addendum, which is included with each testing event with the specimens. Ensure results are reported on the correct line according to method type GHb, HbA1 or HbA1c. Specimens are to be processed as if they were whole (unhemolyzed) blood, even though they will arrive without the cells intact. Samples are not compatible with the NycoCard from Axis Shield. AAB PROFICIENCY TESTING SERVICE ~ 2008 Program Guide 7

10 2008 PROGRAM GUIDE Proficiency Testing Service AAB CHEMISTRY LIPIDS Five liquid serum specimens per testing event for the following analytes: Apolipoproteins A1 and B Lipoprotein (a) LDL Cholesterol HDL Cholesterol Code reagents from the list on the reporting form. Code instrument from the list found in the General Instructions or Addendum, which is included with each testing event. For HDL and LDL, the reagent coded must correspond to the lipoprotein separation reagent, not the cholesterol reagent. Many manufacturers offer both direct and indirect methods. Be careful to select the correct methodology when entering your reagent code. Do not report calculated LDL values. Lipids module is not compatible with the Abaxis Piccalo. PREGNANCY, SERUM or URINE (hcg) Five liquid specimens per testing event. Samples are suitable for both serum and urine pregnancy kits. Only results from vials labeled "Pregnancy" may be reported on the Pregnancy form. Do not report results from the Basic & Comprehensive Chemistry vials on this form. Code your method using the list at the bottom of the Pregnancy reporting form. PTH/INSULIN/C-PEPTIDE Two lyophilized specimens per testing event for the following analytes: C-Peptide PTH Insulin Samples are suitable for both serum and urine pregnancy kits. Reconstitute with 2mLs of distilled water following the instructions given on the reporting form. Code your method using the list at the bottom of the PTH/Insulin/ C-peptide reporting form. SPECIAL CHEMISTRY Two liquid serum specimens per testing event for the following analytes: Ferritin T3, Free Folate Testosterone Homocysteine Vitamin B12 Prolactin Prostate Specific Antigen (PSA), Total Code reagents from the list on the back of the reporting form. Code instruments from the list found in the Addendum, which comes with the specimens. Testosterone Report results only in ng/dl. Multiply ng/ml results by 100 to convert to ng/dl. THERAPEUTIC DRUG MONITORING Five liquid serum specimens per testing event for the following analytes: Acetaminophen Procainamide Carbamazepine Procainamide, N-Acetyl (NAPA) Digoxin Quinidine Gentamicin Salicylates Lithium Theophylline Phenobarbital Tobramycin Phenytoin Valproic Acid Primidone Vancomycin Salicylates Report results in the appropriate line for your reporting units (mg/l or mg/dl). To convert mg/dl to µg/ml, multiply by 10. Note that mg/l and µg/ml are equivalent. Code reagents from the list on the back of the reporting form. Code instrument from the list found in the General Instructions or Addendum, which is included with each testing event. TUMOR MARKERS Two liquid serum vials for analysis of the following analytes: Beta-2-microglobulin CA-125 Carcinoembryonic Prostate-Specific Antigen (CEA) Antigen, (PSA) Free Code your method using the list at the bottom of the Tumor marker reporting form. TIBC/UIBC Two lyophilized serum vials for analysis of the following analytes: TIBC (Total Iron Binding Capacity) UIBC (Unsaturated Iron Binding Capacity) Report the measured parameter only. Do not report both if one result is based solely on a calculation using total iron and the second answer. URINE DRUG SCREENING Two vials of synthetic, liquid urine per testing event for the following analytes: Minimum Target Drugs of Abuse Category Concentrations Alcohol (Ethanol) 0.050% or 50 mg/dl Amphetamines (dl-amphetamine, d-methamptheramine) 1000 ng/ml Barbiturates (Secobarbital) 300 ng/ml Benzodiazepines (Diazepam) 300 ng/ml Cannabinoids (11-nor-delta-9-THC-carboxylic acid) 100 ng/ml Cocaine Metabolite (Benzoylecgonine) 300 ng/ml MDMA (Ecstasy) 500 ng/ml Methadone (Dolophine) 300 ng/ml Methadone Metabolite 100 ng/ml Methaqualone (Quaalude) 3000 ng/ml Methamphetamine 1000 ng/ml Opiates (Morphine) 2000 ng/ml Phenycyclidine (PCP) 25 ng/ml Propoxyphene (Darvon) 300 ng/ml Trycyclic Antidepressants (Amitriptyline) 1000 ng/ml When present, drugs are spiked at greater than the minimum target concentrations listed above. Code method codes from the list on the reporting form. WHOLE BLOOD GLUCOSE Two (Basic) or five (Comprehensive) specimens per testing event. Specimen preparation instructions are displayed on the reporting form(s). Method codes are included on the reporting form(s) and must be reported each time for results to be peer graded. A Single Site form is for reporting only one set of values. A Multiple Site form is for reporting up to 20 sets of values for any combination of different operators and/or methods. However, whether participants are subscribed to a single-site or a multiple-site program, each line of results reported must be derived from the same method for all specimens. When more than one line of results are reported in the multiplesite program, only the first line of values will be scored for CMS reporting purposes, and then only if subscribed to a Comprehensive Whole Blood Glucose (five vial) program. Do not report glucose values in these modules from instruments that generate serum glucose values such as the Abbott Vision or the Roche Reflotron. Serum glucose values are not equivalent to whole blood glucose values. Participants using such methods must subscribe to the serum-based Basic Chemistry program described above. This does not apply to instruments like the Lifescan P/S meters which convert readings to plasma/serum-like values; these meters do require whole blood specimens Program Guide ~ PROFICIENCY TESTING SERVICE AAB

11 AAB Proficiency Testing Service 2008 PROGRAM GUIDE CLINICAL MICROSCOPY & URINALYSIS CLINICAL MICROSCOPY Five photomicrographs per testing event for identification of the following analytes: KOH skin prep Stool Leukocytes Nasal Eosinophils Vaginal Wet Mount Pinworm Prep Code your answers from the list on the bottom on the reporting form. If a photomicrograph demonstrates features which would be referred to another laboratory for identification, report code 10 (would refer). FECAL OCCULT BLOOD Two stool specimens per testing event. Code methods from the lists on the reporting form. URINALYSIS One liquid urine specimen per testing event for the following analytes: Bilirubin Leukocyte Esterase Blood (Hemoglobin) Nitrite Creatinine ph Glucose Protein Ketones Specific Gravity Five photomicrographs per testing event for Sediment Identification. CMS does not monitor urinalysis scores, except for urine pregnancy testing. The liquid specimen is intended only for Urinalysis procedures. Do not use for urine sediment, urine hcg or urine chemistry determinations. Code your qualitative and semi-quantitative answers using only the codes listed on each line of the reporting form. Code your methods from the lists on the reporting form. If using a Bayer instrument, code the instrument rather than the dipstick. Use "ph-corrected" Specific Gravity codes only if you routinely correct for the ph of your specimen; otherwise, use the "ph-uncorrected" codes. Code your Sediment Identification answers from the list on the reporting form. If a photomicrograph demonstrates features which would be referred to another laboratory for identification, report code 10 (would refer). Leave blank if you do not perform Sediment Identification. ACTIVATED CLOTTING TIME Two specimens per testing event. The specimens provided are intended only for activated clotting times, using Actalyke, International Technidyne (ITC), i-stat, Medtronic or Quest Medical ACTester instruments. Code the method used by entering the appropriate numeric code from the list on the reporting form and use only the reconstitution instructions on the reporting form which are specific to your method. COAGUCHEK PT, BASIC Two lyophilized specimens per testing event. Lyophilized samples for analysis of whole blood prothrombin time and INR. These samples are compatible with Roche CoaguChek, CoaguChek S and CoaguChek XS instruments only. Participants with instruments which assay either plasma or whole blood such as Bayer/CVDI TAS, HemoTec ACT or Sienco SonoClot, should enroll in the Coagulation module. Always review the reporting form for instrument specific or updated instructions. Two plastic tubes are provided, each containing an enclosed glass ampule of diluent. Reconstitute and test one vial at a time. Reconstitute each specimen as follows: (1) Allow the samples to come to room temperature at least 20 minutes before reconstituting. (2) Remove the tear strip seal from the tube. (3) Hold the tube upright and tap it 3 to 4 times on a table top to settle the glass ampule to the bottom of the tube. (4) Break the ampule by squeezing the lower half of the tube. The ampule must break. Squeeze the tube with two hands if necessary. Do not bend the tube. This may cause glass shards to cut through the exterior wall of the plastic tube. Do not mix the contents of the tube at this point. (5) Set a timer for 2 1/2 minutes. Again holding the tube upright, tap it hard ten times on a table top or other solid surface to mix the sample. (6) Lay the vial on its side for 2 minutes. Do not move or otherwise disturb the vial during this period. COAGULATION (7) Begin testing by inserting the strip into your Roche CoaguChek monitor. (See reporting form for CoaguChek S specific instructions.) The printed side of the strip must be up and the strip inserted in the direction of the printed arrows. When the monitor displays IS THIS A CONTROL? press YES. (8) The monitor will display PLEASE WAIT. When ready the monitor will display APPLY SAMPLE. (9) Shake the sample hard to force the contents of the tube into the dispenser tip. Hold the tube tip down so that the sample remains in the tip and remove the cap. (10) Save the cap and discard the first drop of sample into it. Then discard both the cap and first drop of sample. (11) Form a fresh drop of sample from the dispenser. Gently touch the drop to the center of the yellow sample target of the test strip. Move the tip around so that the drop completely covers the sample target. Make sure the drop touches the channel surrounding the sample target. (12) As the sample enters the strip test area, the monitor display will change to TESTING and a progress bar will appear. Do not disturb the strip or monitor while the monitor display reads TESTING. Do not add additional sample. (13) Repeat steps 2 through 12 for each additional sample. Code the instrument by entering the appropriate code from the list on the reporting form. COAGUCHEK PT, COMPREHENSIVE Five lyophilized specimens per testing event. Lyophilized samples for analysis of whole blood prothrombin time and INR. These samples are compatible with Roche CoaguChek, CoaguChek S and CoaguChek XS instruments only. Participants with instruments which assay either plasma or whole blood such as Bayer/CVDI TAS, HemoTec ACT or Sienco SonoClot, should enroll in the Coagulation module. Instructions will be printed on the reporting forms. Five plastic tubes are provided, each containing an enclosed glass ampule of diluent. See Coaguchek PT, Basic for reconstitution instructions. AAB PROFICIENCY TESTING SERVICE ~ 2008 Program Guide 9

12 2008 PROGRAM GUIDE Proficiency Testing Service AAB COAGULATION continued COAGULATION, PLASMA Five lyophilized specimens per testing event for the following analytes: Activated Partial Thromboplastin Time (APTT) Fibrinogen INR (for informational use only) Prothrombin Time (PT) Reconstitute these specimens as follows: (1) Add 1.0 ml distilled or deionized water with a volumetric pipet. (2) Invert occasionally until completely dissolved (do not shake). (3) Let stand for 15 minutes after dissolution. (4) Invert again prior to sampling. Once reconstituted, the five coagulation specimens should be processed as though they were fresh citrated patient plasma. If a result's endpoints lack reproducibility, leave the result blank and paper clip a photocopy indicating an indeterminate result. Participants using Roche CoaguChek instruments (which strictly require whole blood) should not use these specimens. Participants using Roche CoaguChek and CoaguChek S instruments may now enroll in the CoaguChek PT program. However, these specimens are compatible with other whole blood methods, including the Abbott Vision, Bayer/CVDI TAS, HemoTec ACT and Sienco SonoClot systems. Code the reagent and instrument used by entering the appropriate codes from the lists provided on the reporting form. All manual coagulation methods must report instrument code 699 (tilt tube) or 700 (wire loop). Fibrinogen Report fibrinogen results only in mg/dl. D-DIMER Two lyophilized specimens per testing event. Code your method using the list at the bottom of the D-Dimer reporting form. Qualitative or semi-quantitative results must be reported as positive/negative. Report any value above the normal range for your assay as positive. Incompatible with Agen SimpliRed WHOLE BLOOD PROTHROMBIN TIME Five specimens per testing event. These specimens are compatible with the Abbott I-Stat. The ITC Microcoag and Roche Coaguchek instruments are NOT compatible with these samples. Participants with instruments which assay either plasma or whole blood such as Bayer/CVDI TAS, HemoTec ACT or Sienco SonoClot, should be enrolled in the Coagulation module. Code the instrument by entering the appropriate code from the list on the reporting form. EMBRYOLOGY, ANDROLOGY & FETAL FIBRONECTIN ANTISPERM ANTIBODIES Two liquid serum specimens per testing event. These specimens are for qualitative IgG evaluation and have been heat inactivated. Store these specimens below -10 C upon arrival. Thaw specimens to room temperature just prior to testing. Perform antisperm antibody testing according to your protocol. Code your answers as either 1 for negative or 2 for positive and code the method used by entering the appropriate numeric code from the list on the form. FETAL FIBRONECTIN (ffn) Two liquid simulated cervicovaginal specimens per testing event. Samples are to be performed in patient mode. Follow your test method s specimen preparation instructions, except do not filter the proficiency test specimen. Report 1 for negative and 2 for positive. IVF EMBRYO CULTURE MEDIA Two culture media specimens per testing event for the following bioassays: Mouse Embryo Human Sperm Survival Hamster Sperm Motility Each vial contains 100 ml of culture media. Also included is 5 ml of dialyzed human serum albumin (100 mg/ml) labeled HSA. Perform your preferred bioassay on both specimens to determine their suitability for in vitro embryo culture. The media should be tested with and without 5 mg/ml HSA. To obtain 5 mg/ml HSA, aseptically add 0.5 ml of the HSA solution to 9.5 ml of each medium and test. SPERM COUNT Two stabilized sperm specimens per testing event. These specimens are for quantitative and qualitative (post-vasectomy) evaluation. Before testing, allow the specimens to warm to room temperature. Vortex for a minimum of 10 seconds or until completely in solution. You may need to repeat this process 3-5 times. Perform the count according to your usual method. Code the method used by entering the appropriate numeric code from the list on the form. SPERM MORPHOLOGY Two unstained slides per testing event. We suggest that you remove the slide labels prior to staining, taking care to reaffix the appropriate label to the corresponding slide after staining. Spermac Users: Spermac fixative for 10 minutes, rinse as per protocol, complete staining protocol. Code the method and stain used by entering the appropriate numeric codes from the list on the form. These samples are not compatible with the TestSimplet method. SPERM VIABILITY Two Eosin-Nigrosin stained slides per testing event. Perform the sperm viability assessment according to your usual method Program Guide ~ PROFICIENCY TESTING SERVICE AAB

13 AAB Proficiency Testing Service 2008 PROGRAM GUIDE CELL IDENTIFICATION Refer to the Blood Cell Photomicrographs result code list on the reporting form, and code a single answer for the cell(s) or cellular feature(s) in each transparency, without regard to results obtained from any of the blood specimens. Photomicrographs should be associated only with the CBC data provided. If identification of abnormal cell(s) or cellular feature(s) at the end of the arrow(s) would routinely be referred, report code 100 (Abnormal, would refer). Participants should not use abnormalities in the clinical data as justification for reporting code 100. This code will be flagged as incorrect if it is reported for normal cells or normal cellular features. CENTRIFUGAL HEMATOLOGY Five whole blood specimens per testing event for the following analytes: Hematocrit Lymph & Monos Leukocytes Platelets Granulocytes Hemoglobin These specimens are only compatible with the Quantitative Buffy Coat (QBC) series of instruments, except the QBC HemaScan. Participants using older models in this series of instruments (which do not generate hemoglobin results) may report hemoglobin results on these specimens using the methods listed in the hemoglobin method code list. Specimen preparation instructions and method codes are included on the reporting form. You must report these codes to have your results graded according to the tube and/or instrument used. Please test as soon as possible after receipt of the specimens, within 5 days if possible. ERYTHROCYTE SEDIMENTATION RATE Two specimens per testing event. Allow the samples to stand at room temperature for 20 minutes prior to use. Gently invert the vial until packed cells have been re-suspended and continue mixing for 30 seconds. Avoid foaming and do not vortex. Follow the manufacturer's directions for filling the sample tubes. These specimens are for manual and automated methods. Round all results to whole numbers to fit the reporting format provided. Code the method used by entering the appropriate numeric code from the list on the reporting forms. Micro-ESR methods use tubes with 1.0 mm internal diameters. Westergren methods use tubes with approximate 30 cm length and 2.5 mm internal diameter. Wintrobe methods use tubes with approximate 10 cm length and 2.5 mm or larger internal diameters. Check the manufacturer's package insert to verify the type of method to report. If the sample is diluted with saline prior to analysis, choose a diluted method for reporting results. ERYTHROCYTE SEDIMENTATION RATE-RAPID Two specimens per testing event. These specimens are for rapid ESR test systems only. Instructions and method codes are included on the reporting form. Compatible with Polymedco SediMat 15. HEMATOLOGY Five whole blood specimens per testing event for the following analytes: Leukocytes (WBC) Hematocrit Erythrocytes (RBC) Platelets Hemoglobin The specimens must reach room temperature (15-30 C) before being resuspended. To resuspend contents, roll the vials between the palms of the hands for 20 seconds in upright, then inverted positions. Place the vials on a mechanical mixer or gently invert manually for 5 minutes. These specimens are not intended for use with the Becton- Dickinson QBC instruments. (QBC users must enroll in the Centrifugal Hematology module.) HEMATOLOGY Incompatible Instruments The following instruments have no compatible differential program and should be enrolled in the Hematology program with cell ID: Bayer H1, H2, H3, Advia 60, 70 and 120 Automated Instrument Issues Participants may experience "backlit" white counts or "R" codes; without any other indications of specimen degradation. Participants need not be reluctant to report these results. ABX 8000 participants may have to use the "print 55" code to report leukocyte counts on these specimens. In the interests of uniformity and accurate grading data, Bayer H-series and Advia users must always report the basophil channel (the "WBC-B" counts) when reporting leukocyte results, regardless of their reporting protocols for patient results. Manual Method Issues All manual cell counting participants must report method code 697. For manual leukocyte counts, use a Unopette system containing acetic acid or ammonium oxalate as diluent; erythrocyte ghosts may be present, but the human leukocytes are easily discernible. For manual platelet counts, use only ammonium oxalate as diluent and count only the mock platelets ("footballs" of uniform size and shape); do not seek and do not count anything which appears to be true platelets (there are no actual platelets in these specimens). For spun hematocrits, ignore the pink supernatants, which are characteristic of these stabilized specimens and will not contribute to out-of-range results. Also, all manual hematocrit participants must choose between method codes 699, 885 or 969. HEMATOLOGY w/diff A Five whole blood specimens per testing event for the following analytes: Leukocytes (WBC) Platelets Erythrocytes (RBC) Lymphocyte Hemoglobin Md/Mid/Mixed/Mono/Other Hematocrit Neutrophils/Granulocytes Five photomicrographs per testing event for Cell Identification. These specimens are compatible with the following instruments: Abbott Cell-Dyn 610, 1200, 1400, 1500, 1600, 1700, 1800 ABX Argos, Helios, Minos, Stx/Stel/Stex Careside all 3-part diff instruments Coulter Ac T series (excluding Ac T-5 & -8), J series, MD series, Onyx, S Plus series, ST series, T series DANAM/Infolab all 2-part diff models Drew Evolution Elan all 3-part diff instruments Medionic all 3-part diff instruments Nova Celltrak The specimens must reach room temperature (15-30 C) before being resuspended. To resuspend contents, roll the vials between the palms of the hands for 20 seconds in upright, then inverted positions. Place the vials on a mechanical mixer or gently invert manually for 5 minutes. Participants with 1-part diff instruments use only the first line designated "Lymphocytes" in the differential section of their forms. Participants with 2-part diff instruments must not use the second line designated "MD/MID/Mixed/Mono/Other" in the differential section of their forms; these participants must report one part on the "Lymphocytes" line and the remaining part on the "Granulocytes" line, regardless of more inclusive terms on their instruments (such as "MO+GR"). Some participants have instruments that report specific granulocyte results (eosinophils and basophils) in addition to the total granulocyte count. This type of reporting is neither appropriate nor expected of participants in this module. Only participants enrolled in Diff B, C and E modules are required to report 5-part differentials. Participants may experience "backlit" white counts or "R" codes; without any other indications of specimen degradation, participants need not be reluctant to report these results. AAB PROFICIENCY TESTING SERVICE ~ 2008 Program Guide 11

14 2008 PROGRAM GUIDE Proficiency Testing Service AAB HEMATOLOGY continued HEMATOLOGY w/diff B Five whole blood specimens per testing event for the following analytes: Leukocytes (WBC) Neutrophils Erythrocytes (RBC) Lymphocytes Hemoglobin Monocytes Hematocrit Eosinophils Platelets Basophils These specimens are compatible with the following instruments: Abbott Cell-Dyn 3000, 3200, 3500, 3700, 4000, Saphire, Ruby DANAM Excell 22 The specimens must reach room temperature (15-30 C) before being resuspended. To resuspend contents, roll the vials between the palms of the hands for 20 seconds in upright, then inverted positions. Place the vials on a mechanical mixer or gently invert manually for 5 minutes. HEMATOLOGY w/diff C Five whole blood specimens per testing event for the following analytes: Leukocytes (WBC) Neutrophils Erythrocytes (RBC) Lymphocytes Hemoglobin Monocytes Hematocrit Eosinophils Platelets Basophils These specimens are compatible with the following instruments: Coulter Gen-S, HMX, LH500, LH750, MAXM, STKS (the VCS series) These specimens must reach room temperature (15-30 C) before being resuspended. To resuspend contents, roll the vials between the palms of the hands for 20 seconds in upright, then inverted positions. Place the vials on a mechanical mixer or gently invert manually for 5 minutes. HEMATOLOGY w/diff D Five whole blood specimens per testing event for the following analytes: Leukocytes (WBC) Platelets Erythrocytes (RBC) Lymphocytes Hemoglobin Md/Mid/Mixed/Mono/Other Hematocrit Neutrophils/Granulocytes These specimens are compatible with the following instruments: Sysmex E series, F-800, K series (includes the KX-21), Poochi The specimens must reach room temperature (15-30 C) before being resuspended. To resuspend contents, roll the vials between the palms of the hands for 20 seconds in upright, then inverted positions. Place the vials on a mechanical mixer or gently invert manually for 5 minutes. Participants with 2-part diff instruments must not use the second line designated "MD/MID/Mixed/Mono/Other" in the differential section of their forms; these participants must report one part on the "Lymphocytes" line and the remaining part on the "Granulocytes" line, regardless of more inclusive terms on their instruments (such as "MO+GR"). HEMATOLOGY w/diff E Five whole blood specimens per testing event for the following analytes: Leukocytes (WBC) Neutrophils Erythrocytes (RBC) Lymphocytes Hemoglobin Monocytes Hematocrit Eosinophils Platelets Basophils These specimens are compatible with the following instruments: Sysmex NE series, SE series, SF series, XE series, XS 1000i, XT series The specimens must reach room temperature (15-30 C) before being resuspended. To resuspend contents, roll the vials between the palms of the hands for 20 seconds in upright, then inverted positions. Place the vials on a mechanical mixer or gently invert manually for 5 minutes. Process specimens in the QC ( Control Key ) mode. HEMATOLOGY w/diff F Five whole blood specimens per testing event for the following analytes: Leukocytes (WBC) Platelets Erythrocytes (RBC) Lymphocyte Hemoglobin Md/Mid/Mixed/Mono/Other Hematocrit Neutrophils/Granulocytes These specimens are compatible with the following instruments: ABX all 3-part diff models excluding Argos, Helios, Minos, Stx/Stel/Stex (see Hematology w/diff A) The specimens must reach room temperature (15-30 C) before being resuspended. To resuspend contents, roll the vials between the palms of the hands for 20 seconds in upright, then inverted positions. Place the vials on a mechanical mixer or gently invert manually for 5 minutes. Participants with 1-part diff instruments use only the first line designated Lymphocytes in the differential section of their forms. Participants with 2-part diff instruments must not use the second line designated "MD/MID/Mixed/Mono/Other" in the differential section of their forms; these participants must report one part on the "Lymphocytes" line and the remaining part on the "Granulocytes" line, regardless of more inclusive terms on their instruments (such as "MO+GR"). Some participants have instruments which report specific granulocyte results (eosinophils and basophils) in addition to the total granulocyte count. This type of reporting is neither appropriate nor expected of participants in this module. Only participants enrolled in Diff B, C and E modules are required to report 5-part differentials. Participants may experience "backlit" white counts or "R" codes; without any other indications of specimen degradation, participants need not be reluctant to report these results. HEMATOLOGY w/diff G Five whole blood specimens per testing event for the following analytes: Leukocytes (WBC) Neutrophils Erythrocytes (RBC) Lymphocytes Hemoglobin Monocytes Hematocrit Eosinophils Platelets Basophils These specimens are compatible with the following instruments: ABX Pentra series 60C+, 80, 120, Coulter Ac T-5, Ac T-8 The specimens must reach room temperature (15-30 C) before being resuspended. To resuspend contents, roll the vials between the palms of the hands for 20 seconds in upright, then inverted positions. Place the vials on a mechanical mixer or gently invert manually for 5 minutes. Process specimens in the QC ( Control Key ) mode. Ω Program Guide ~ PROFICIENCY TESTING SERVICE AAB

15 AAB Proficiency Testing Service 2008 PROGRAM GUIDE RETICULOCYTE COUNT Two specimens per testing event. Specimens for manual and automated instruments. Samples are compatible with the following instruments: Abbott Cell-Dyn 3200, 3500, 3700, 4000 Beckman Coulter HMX, LH, XL, STKS, MAXM Becton Dickinson FACS Series Samples are not compatible with the: Beckman Coulter Gen-S HEMATOLOGY continued IMMUNOHEMATOLOGY Reconstitute these specimens as follows: (1) Refrigerate specimens upon arrival. Even if specimens are to be used that day allow specimens to chill at least 3 to 4 hours before use. The specimens should be tested immediately after removing from the refrigerator. Specimens must be tested cold. (2) Mix specimens by gentle inversion between thumb and index finger until the red blood cells are completely resuspended. Do not mix mechanically. Do not rub between palms of hands. (3) Perform testing as you would patient samples. BASIC IMMUNOHEMATOLOGY Five whole blood specimens per testing event for the following analytes: ABO Group D (Rho) Typing These specimens should be processed as you would patient material. The supernatant of each specimen is used for reverse typing. Code all your answers using only the numbers available on the reporting form. Code your method from the list on the form. Note: You do not need to wash the cells to do front typing. D (Rho) Typing Laboratories not performing the weak D (D u ) test on D (Rho)-negative specimens must report code 1 for negative, weak D (D u ) not performed. Laboratories performing the weak D (D u ) test on D (Rho)-negative specimens must report either code 2 for negative or code 4 for weak D (D u )-positive. If code 2 (negative) is reported, it is assumed the weak D test was performed and should the specimen be a weak D, a flag representing an incorrect answer will result. COMPREHENSIVE IMMUNOHEMATOLOGY Five whole blood (simulated donor) and five serum (simulated recipient) specimens per testing event for the following analytes: ABO Group D (Rho) Typing Unexpected Antibody Identification Unexpected Antibody Detection Compatiblity Testing (Crossmatch) Both types of specimens should be processed as you would patient material except as indicated below. You do not need to wash the cells to do front typing. For ABO/Rh, use the cells for forward typing and the supernate above the cells for reverse typing; for compatibility and antibody detection/id, use the sera. Code all your answers using only the numbers available on the reporting form. Code the method from the list on the form. Code only the method you use to confirm negative reactions. D (Rho) Typing Laboratories not performing the weak D (D u ) test on D (Rho)-negative specimens must report code 1 for negative, weak D (D u ) not performed. Laboratories performing the weak D (D u ) test on D (Rho)-negative specimens must report either code 2 for negative or code 4 for weak D (D u )-positive. If code 2 (negative) is reported, it is assumed the weak D test was performed and should the specimen be a weak D, a flag representing an incorrect answer will result. Compatibility Testing Perform a full crossmatch of each serum specimen labeled Unexpected Antibodies with the like-numbered ABO/Rh whole blood specimens. Results will be graded on the basis of serological compatibility, not whether the blood would actually be released for transfusion, a decision which often depends on clinical circumstances and local protocol. Unexpected Antibody Detection and Identification Use vials labeled Unexpected Antibodies and perform antibody screening or identification as you would patient sera. If you are unable to identify the antibody, report the ID as code 10-would refer. For 10 and 20 cell panel identification, please order additional specimens. DIRECT ANTIGLOBULIN TEST Two specimens per testing event. Suspension of red blood cells for direct antiglobulin testing. Instructions and method/instrument codes are included in your reporting form. IMMUNOLOGY ANTINUCLEAR ANTIBODY Five liquid serum specimens per testing event. Participants may report results from any of three types of methods: classical (IEA/IFA), anti-dnp (LA) or anti-ena (EIA); but they must not report results from more than one method in the same testing event. Code either the absence or presence of antinuclear antibody, using either 1 for negative (or for normal patterns in the case of IEA/IFA methods) or 2 for positive (or for abnormal patterns in the case of IEA/IFA methods). Code the method used by entering the appropriate numeric code from the corresponding list on the reporting form. ANTISTREPTOLYSIN O Five liquid serum specimens per testing event. Code your qualitative answers using only the codes provided on the "Antistreptolysin O" line of the Diagnostic Immunology reporting form and code the method used by entering the appropriate numeric code from the list on the form. C-REACTIVE PROTEIN One liquid serum specimen per testing event. Code your qualitative answers using only the codes provided on the "C-Reactive Protein qualitative" line of the Diagnostic Immunology reporting form and code the method used by entering the appropriate numeric code from the list on the form. Quantitative results are to be entered on the subsequent line. HELICOBACTER PYLORI ANTIBODY Two liquid serum specimens per testing event. These specimens are also designed to work with whole blood methods, but participants must use their method's collection tubes just as they would for testing patients. Fill the collection tube directly from the specimen vial, then follow your routine testing protocol. Code your qualitative answers as either 1 for negative or 2 for positive and code the method used by entering the appropriate numeric code from the list on the form. AAB PROFICIENCY TESTING SERVICE ~ 2008 Program Guide 13

16 2008 PROGRAM GUIDE Proficiency Testing Service AAB IMMUNOLOGY continued HEPATITIS MARKERS Five liquid serum specimens per testing event for the following analytes: anti-hbc (IgG+IgM or IgM) HBsAg anti-hbs anti-hcv HBeAg Code your qualitative answers only with the result codes on each line of the reporting form and code your methods using the reagent codes included on the form. If your method is immunoglobulin (Ig)-nonspecific or its specificity is not known, report results on the IgG line. Special Safety Precautions Specimens testing positive for anti-hiv have been heat treated. Even so, all these specimens should be handled as though they are capable of transmitting disease. Follow CDC and FDA recommendations as well as the Occupational Safety and Health Administration (OSHA) Final Rule on Occupational Exposure to Bloodborne Pathogens published December 6, 1991 (Federal Register Volume 56, Number 235, pages ). These specimens should be handled in a biological safety cabinet while wearing impermeable gloves. Remove the specimens from the can using forceps that are long enough to avoid cutting or scratching your hands on the can. Cover the caps of the vials with gauze while removing or replacing to prevent splashing. Do not pipet by mouth or use a needle and syringe with these specimens. Wash your hands after removing the gloves. These specimens should be autoclaved and disposed of as hazardous waste. HIV MARKERS Five liquid serum specimens per testing event for the following analytes: anti-hiv-1 or 1/2 Screening anti-hiv-1 Confirmation anti-hiv-2 Screening anti-hiv-2 Confirmation These specimens are compatible for serum, plasma or whole blood methods. Code your qualitative answers only with the result codes on each line of the reporting form and code your methods using the reagent codes included on the form. To avoid false positive results, participants using membrane-based methods must centrifuge specimens prior to testing. Screening and/or confirmatory results may be reported for anti-hiv-1/anti-hiv-2, but only one method can be used for CMS subspecialty scoring in general immunology, the one pre-designated as such by participants, which, according to CMS, should be your routine (highest volume) method. Since scoring discrepancies may occur between methods, ensure that you report all five results for any of these HIV markers from the same method, whether your routine method is a screening method or a confirmatory method. Those who report confirmatory (rather than their screening) anti-hiv results that may not routinely perform confirmatory testing on patient specimens which are negative on initial screen, per CMS requirements must report results on all five HIV Marker specimens to avoid being penalized for missing results. If you do not routinely perform confirmatory testing on negative specimens, please use the result code indicating that this is the case. Special Safety Precautions Specimens testing positive for anti-hiv have been heat treated. Even so, all these specimens should be handled as though they are capable of transmitting disease. Follow CDC and FDA recommendations as well as the Occupational Safety and Health Administration (OSHA) Final Rule on Occupational Exposure to Bloodborne Pathogens published December 6, 1991 (Federal Register Volume 56, Number 235, pages ). These specimens should be handled in a biological safety cabinet while wearing impermeable gloves. Remove the specimens from the can using forceps that are long enough to avoid cutting or scratching your hands on the can. Cover the caps of the vials with gauze while removing or replacing to prevent splashing. Do not pipet by mouth or use a needle and syringe with these specimens. Wash your hands after removing the gloves. These specimens should be autoclaved and disposed of as hazardous waste. hscrp Two liquid serum specimens per testing event. Quantitative results are to be entered on the hscrp reporting form. Code the method used by entering the appropriate code from the list on the form. IMMUNOPROTEINS Five liquid serum specimens per testing event for the following analytes: C3 IgE C4 IgG IgA IgM Where appropriate, all participants must report results which are calibrated against the International Federation of Clinical Chemistry (IFCC)'s Reference Preparation for Proteins in Human Serum (RPPHS). Code the method(s) used by entering the appropriate numeric codes from the list(s) on the form. INFECTIOUS MONONUCLEOSIS Five liquid serum specimens per testing event. These specimens are for infectious mononucleosis (IgG or IgM) procedures. Code your qualitative answers using only the codes provided on the "Infectious Mononucleosis" line of the Diagnostic Immunology reporting form and code the method used by entering the appropriate numeric code from the list on the form. LYME DISEASE Two liquid serum specimens per testing event. Be careful to enter results on the proper line (lines are provided for both screening and immunoblot results). Code your qualitative answers as either 1 for negative or 2 for positive and code the method used by entering the appropriate numeric code from the list on the form. ORAL FLUID HIV ANTIBODIES Five oral fluid vials per testing event. Submit the contents of these vials directly to your routine testing system. Western Blot may be performed on these specimens. The presence of any band along the prerequisite bands should be interpreted as positive. RHEUMATOID FACTOR Five liquid serum specimens per testing event. These specimens are for qualitative rheumatoid factor (IgG or IgM) procedures. Code your qualitative answers using only the codes provided on the "Rheumatoid Factor" line of the Diagnostic Immunology reporting form. For quantitative methods, report your results as greater than or less than your methods cutoff. Code the method used by entering the appropriate numeric code from the list on the form. Follow the manufacturer's instructions for the pretreatment of serum specimens just as you would for any patient specimen Program Guide ~ PROFICIENCY TESTING SERVICE AAB

17 AAB Proficiency Testing Service 2008 PROGRAM GUIDE CONTINUED IMMUNOLOGY RUBELLA Five liquid serum specimens per testing event. Be careful to enter results on the proper line (lines are provided for both IgG and IgM results). If your method is immunoglobulin (Ig)-nonspecific or its specificity is not known, report results on the IgG line. Code your answers as either 1 (not immune or negative) or 2 (immune or positive). If using a quantitative method, use the upper cutoff value of your normal range to distinguish not immune from immune status. Code the method used by entering the appropriate code from the list provided on the form. SYPHILIS SEROLOGY Five liquid serum specimens per testing event. Code your answers using only the codes listed on the line of the reporting form which is appropriate to the method used. Code the methods used by entering the appropriate codes from the list provided on the form. Separate lines are provided to accommodate participants' results from more than one method. Procedures for which results are preformatted include RPR Card, RST, TRUST, VDRL, USR, FTA-ABS, FTA-ABS DS, MHA-TP/TP-PA, and EIA, but only one method can be used for CMS subspecialty scoring in syphilis serology, the one pre-designated as such by participants. Do not attempt to use the same line to report results from more than one procedure. Participants who request that we report their confirmatory (rather than their screening) syphilis results to the regulatory authorities, even those who may not routinely perform confirmatory testing on patient specimens which are negative on a screening procedure, should report results on all five Syphilis specimens to avoid being penalized for missing results. MICROBIOLOGY ACID-FAST SMEARS Five specimens per testing event. These specimens will only be included in the first and third shipments. The specimens are killed bacteria suspended in simulated sputum. Perform your normal staining procedures and code either the absence or presence of acid-fast bacilli, using only the numbers 1 (for acid-fast bacilli not present) or 2 (for acid-fast bacilli present) and code the method used by entering the appropriate numeric code 1 (for nonfluorescent) or 2 (for fluorescent). The Acid-fast Smears program is limited to Mycobacteriology Type 1 (Extent 1) laboratories only (those which only report acid-fast smear results). Therefore, extent coding is not relevant for this program. AFFIRM MICROBIAL SCREEN Five Affirm Nucleic Acid Probe Assays for analysis of the following: Gardnerella Candida Trichomonas Code the reagent and instrument used by entering the appropriate codes from the lists provided on the reporting form. BACTERIOLOGY GC CULTURE THROAT CULTURE THROAT/URINE CULTURE URINE CULTURE Five specimens per testing event. Inspect the fiberboard container carefully for all inclusions. Five specimens are provided for each culture program. After inventorying the contents of the container, store them in a refrigerator. Special Safety Precautions These specimens contain pathogens or potential pathogens and should be considered infectious and handled as though they are capable of transmitting disease. They should be handled and disposed of only by personnel trained to work with pathogenic bacteria. All laboratory precautions and safety measures appropriate to handling live cultures should be practiced when working with these specimens. In addition to the Precautions section on page 3, one should be especially careful to avoid aerosol creation, inhalation, ingestion or injection of bacteria. These specimens should be autoclaved and disposed of as hazardous waste. Instructions for Culture Specimens One swab, lyophilized sample pellet and rehydration fluid is provided for each specimen (1) Remove the tube from the outside packaging. No warming is required, samples may be used cold. (2) Remove the cap that holds the swab and inoculate media. (3) For single plate inoculation: For heavy growth, rotate the swab while streaking the entire plate in all quadrants. For isolated colonies, rotate the swab over the first quadrant only. Utilize a sterile loop to streak the remaining quadrants as per your normal laboratory procedure. (4) For multiple plates or Uricult users: Submerge the swab portion only into the provided rehydration broth for 15 to 30 seconds. Remove the moist swab and use it to streak plates as per your laboratory s procedure. The broth may be capped and stored as you would any other stock broth. Uricult users should use the inoculated broth to wet the test paddle. Determining Type (Extent) of Laboratory Service All participants, regardless of the extent of their laboratory practice, evaluate the same specimens. In order to be graded appropriately, you must report the extent of laboratory practice (Extent 0, 1, 2, 3, 4 or 5) for each specimen. Determine your extent as follows: (1) Subject each specimen to your protocol for each source as described at each specimen number on your reporting form. (2) Based on what you would report in the context of your specific laboratory practice, determine your extent for each specimen independently of each other, according to the definitions on the reporting form and clarified in the following table: Results Extent of Laboratory Service Reported Gram must must may may may may Stain not rpt report report report report report Antigen must may must may may may Screen not rpt report report report report report Antimicrobial Suscept- may may may may may may ibility Testing (ASTs)* report report report report report report Identification must may may must must must to Genus Only not rpt report report report not rpt not rpt Speciation must may may may must must of Aerobes not rpt report report report report report Identification must may may may may must of Anaerobes not rpt report report report report report *Reporting ASTs applies only to Specimen 1 and assumes the use of pure isolates. Participants performing ASTs without identifications, even presumptive ones, must use Extent 0 to avoid being given a score of zero for missing culture results. (3) Participants that report would refer on all five culture samples, will receive a DC (discontinued testing) on their graded report for that testing event. AAB PROFICIENCY TESTING SERVICE ~ 2008 Program Guide 15

18 2008 PROGRAM GUIDE Proficiency Testing Service AAB MICROBIOLOGY continued (4) Regardless of the extent indicated, we are required to grade the most definitive identification reported. This means, for example, that a species result takes precedence in grading over a genus, antigen or Gram stain result. (5) If unable to decide between extents for a given specimen, report the lowest extent which applies, except Extent 0. In order to preserve the opportunity to get a zero on one specimen and still receive 80 percent overall, it is advisable to avoid using Extent 0 when Gram stain (Extent 1) or antigen screening (Extent 2) results are available for reporting. It is acceptable to challenge your Gram stain or antigen screening procedure(s) with our bacteriology specimens (without regard to specimen source information) in order to receive five specimen scores upon which to generate an overall procedure score for bacterial identifications. (6) Even though, Extent 4 laboratories are not required to identify anaerobes, they must detect their presence in appropriate cultures (i.e. wound, stool, blood). If the laboratory does not test for the presence of anaerobes in these samples, do not select any extent higher than Extent 3. Failure to enter an extent will result in your laboratory being evaluated as Extent 5. This cannot be corrected once grading has occurred. Coding for Presumptive Culture Identification Participants reporting presumptive identifications by culture must not use result codes 140 to 616. If performing isolations only with selective media, these participants might need to report code 697 (No pathogen found), but should not report either code 698 (No aerobic growth) or 699 (No aerobic or anaerobic growth). CMS requires that we challenge many common pathogens found in a specific sample type. When reporting a negative result, select an answer which reflects the organisms you would normally detect (I.e. select code 667 if you only screen for Strep A and do not test for N. gonorrhoeae with throat cultures.) Coding Extent 3, 4 and 5 Results For Specimens 1, 2, 3 and 4, participants using Extent 3, 4 or 5 must report only the organism(s) which they consider to be the significant pathogen(s) that is/are clearly responsible for the illness described, excluding immunocompromised patients. Opportunistic pathogens occurring in immunocompromised patients, when included, will always appear in Specimen 5. All organisms, nonpathogens as well as pathogens, must be identified in Specimen 5. Code your answers using the Result Code list on the reporting forms. Antimicrobial susceptibility tests (ASTs) are to be performed on the most significant pathogen in Specimen 1 only, using the AST codes listed on the reverse side of reporting forms. Caution: if you code extent 0 or 1 (culture ID referred) for Specimen 1 and want to be exempt from reporting ASTs on this specimen, you must also code your AST method as 0 ("susceptibilities not routinely performed on this organism") and leave the AST results blank. Due to new CMS requirements, participants will be flagged for inappropriate selection of AST s, as listed in the CLSI (formerly NCCLS) guidelines M2-A8 and M7-A6. Selection of an inappropriate AST will be counted as an incorrect response. Refer to your reporting form for specimen sources and other reporting requirements. C. DIFFICILE TOXIN ANTIGEN DETECTION Five lyophilized specimens per testing event. Special Safety Precautions These specimens contain pathogens or potential pathogens and should be considered infectious and handled as though capable of transmitting disease. Specimens should be handled and disposed of by personnel trained to work with pathogenic organisms. All laboratory precautions and safety measures appropriate to handling live cultures should be practiced when working with these specimens. In addition to the Precautions on page 3, one should be especially careful to avoid aerosol creation, inhalation, ingestion or injection of organisms. These specimens should be autoclaved and disposed of as hazardous waste. If the kit contains broken specimens, autoclave the kit with minimal exposure of the contents to the atmosphere and immediately request replacement samples. Specimen Analysis Resuspend each specimen by adding 1.0 ml of sterile distilled water. Use a sterile syringe and needle to add the water into the vial. Gently swirl the vial to dissolve the contents. Allow 1-3 minutes for the contents to completely dissolve. Process the specimens according to your kit manufacturer s instructions. Report 1 for negative and 2 for positive. Code the instrument by entering the appropriate code from the list on the reporting form. CHLAMYDIA/GC/STREP GROUP B ANTIGEN SCREEN Five antigen suspension specimens per testing event for the following analytes: Chlamydia trachomatis Antigen Screen Neisseria gonorrhoeae Antigen Screen Streptococcus sp., Group B Antigen Screen Each specimen may contain one or more bacteria, but no attempt should be made to culture or stain these specimens. Special Safety Precautions These specimens contain pathogens or potential pathogens and should be considered infectious and handled as though they are capable of transmitting disease. They should be handled and disposed of only by personnel trained to work with pathogenic bacteria. All laboratory precautions and safety measures appropriate to handling live cultures should be practiced when working with these specimens. In addition to the Precautions section on page 3, one should be especially careful to avoid aerosol creation, inhalation, ingestion or injection of bacteria. These specimens should be autoclaved and disposed of as hazardous waste. Specimen Analysis Follow the instructions for reconstitution as indicated below: (1) Pour the liquid into the vial containing the pellet. (2) Resuspend gently by flicking the vial. (3) Use immediately after the pellet has dissolved. (4) Refrigerate any unused portion. It should be stable for at least 24 hours. Use the swab to absorb a portion of the rehydrated sample. If testing more than one antigen, be sure to apportion the specimen accordingly (about 2 drops per swab is sufficient). For users of gender specific methods, this sample is intended to be used with the female collection kit. Do not omit inoculation of transport media with the wetted swab if this step is applicable to patient material. Follow your manufacturer s recommendations for this final step. If the manufacturer s directions for incubation time in the transport media are not clear, please allow a minimum of 30 minutes. Code your answers as either 1 for negative or 2 for positive. Code the method(s) used by entering the code(s) from the appropriate list(s) of methods provided on the reporting form. Specimens compatible with the DNA probe test kit. CRYPTOSPORIDIUM/ GIARDIA IMMUNOASSAY Five formalin specimens per testing event for the following analytes: Cryptosporidium Immunoassay Giardia Immunoassay Perform your normal procedure and code results as 1 for negative and 2 for positive. Code the method used by entering the appropriate code from the list on the form. These samples formalin preserved and are not compatible with methods requiring fresh specimens Program Guide ~ PROFICIENCY TESTING SERVICE AAB

19 AAB Proficiency Testing Service 2008 PROGRAM GUIDE CONTINUED MICROBIOLOGY GRAM STAIN Five unstained samples per testing event. Perform your normal staining procedure, including fixing of smears, and code your answers as either 1 (gram-negative) or 2 (gram-positive). PARASITOLOGY Five specimens per testing event. Specimens 1, 2 and 3 are formalin preserved feces for direct examination and concentration procedures. The like-numbered PVA slide is for staining and is either a replicate or will contain the same organism(s) as the formalinized specimen. Occasionally, blood smears will be substituted for fecal specimens. For participants using stool fixatives that contain a mercuric chloride substitute (zinc sulfate, etc.), remember that the proficiency testing specimens you receive for permanent staining have been preserved in PVA using the mercuric chloride fixative base. If you use the Trichrome or iron hematoxylin staining method for your mercuric chloride substitute fixatives, you may have eliminated the 70% alcohol/iodine step and the following 70% alcohol rinse step from your method. However, when you stain the proficiency testing fecal smears, you will need to incorporate the iodine step plus the next 70% alcohol rinse back into your staining protocol prior to placing your slides into the Trichrome stain or iron hematoxylin stain. These two steps are designed to remove the mercury from the smear and then to remove the iodine; therefore, when your slide is placed into the Trichrome or iron hematoxylin stain, both the mercury and iodine are no longer present in the fecal smear. If you fail to incorporate these two steps into your staining protocol, the quality of your proficiency testing stained smears will be poor. Specimens 4 and 5 are photomicrographs (trans-parencies); therefore, do not report frequencies on specimens 4 and 5. Report the extent of your parasitology practice for each specimen based on the definitions printed on the reporting form. Participants that report would refer on all five samples, will receive a DC (discontinued testing) on their graded report for that testing event. Code your answers with the Parasitology Organism Codes listed on the reporting form. Code 98 [parasite(s) found, not identified] is appropriate only for Extent 1 specimens. Laboratories receiving specimens which are not routinely processed (such as blood smears) should report Extent 0 (laboratories which do not process or would refer specimen). We are required to categorize participants who fail to report their extent(s) as Extent 2 and to grade accordingly. STREP GROUP A ANTIGEN SCREEN Five specimens per testing event. Specimens are inoculated with killed bacteria and therefore no attempt should be made to culture or stain these specimens. Participants reporting results of bacterial culture or staining techniques must subscribe to the Bacteriology, GC Culture, Gram Stain, Throat Culture, Throat/Urine Culture or Urine Culture programs. DNA Probe methods or any other method requiring live bacteria should be tested with the Throat Culture program. In performing a rapid Group A antigen screen on these specimens, follow the same procedure used on patient specimens. If you use the Quidel QuickVue In-Line product or the BioStar OIA method then you must follow special instructions for proficiency testing found in the kit's package insert. BioStar OIA participants must pre-wet each swab with 3-4 drops of Reagent 4 (Wash Solution) or sterile saline prior to testing. If the specimen's fluid content is inadequate for other procedures, add sterile saline (generally 1-2 drops) to the swab and express the fluid. Code your answers as either 1 for negative or 2 for positive. Code the method used by entering the appropriate code from the list on the form. VIRAL ANTIGEN SCREEN Five specimens per testing event for analysis of the following analytes: Influenza Type A Influenza Type B Respiratory syncytial virus (RSV) Antigen Rotavirus Antigen These samples are for non-if antigen detection methods only. Special Safety Precautions These specimens contain pathogens or potential pathogens and should be considered infectious and handled as though they are capable of transmitting disease. They should be handled and disposed of only by personnel trained to work with pathogenic bacteria. All laboratory precautions and safety measures appropriate to handling infectious materials should be practiced when working with these specimens. In addition to the Precautions section on page 3, one should be especially careful to avoid aerosol creation, inhalation, ingestion or injection of bacteria. These specimens should be autoclaved and disposed of as hazardous waste. Specimen Analysis Resuspend each specimen by adding 1.0 ml of sterile distilled water. Use a sterile syringe and needle to add the water into the vial. Gently swirl the vial to dissolve the contents. Allow 1-3 minutes for the contents to completely dissolve. Process the specimens according to your kit manufacturer s instructions. Follow the manufacturer s procedure for running a positive control for the following methods. BioMerieux Slidex Rota-Kit 2, Murex Rotavirus Latex, Meridian ImmunoCard, Meritec Rotavirus, or Virogen Rototest. Code your answers using the result codes on the reporting form. Code your method(s) used by entering the appropriate code(s) from the lists on the reporting form. CLIA requires a total of 5 viral antigen challenges. Be sure to test all 5 samples. Partial results or blanks will be scored as misses. Your accrediting agency may require you to perform proficiency testing for all antigen tests done in your laboratory. GRADING PROCEDURES INTRODUCTION Grading procedures must conform to the February 28, 1992 Final Rule which implements the CLIA '88. This rule retains two practices previously mandated regulations from CMS: (1) The principle of peer grading (participant consensus) determines target values (satisfactory performance), especially for quantitative results. (2) The principle of medical (fixed) performance standards primarily determines grading limits, but not to the total exclusion of the principle of methodological (statistical) performance standards. The major aspects of grading procedures are detailed to follow. CONSENSUS GRADING The grading of all results depends on the existence of a consensus of acceptable results. For most analytes or procedures, CMS defines consensus as acceptable performance among 80% or more of results from all participants or referees. Per CMS guidelines, if 80% consensus Is achieved among all laboratories or referee laboratories, then the analyte must be graded. Exceptions to this rule exist for all CMS-scored procedures in Immunohematology, in which consensus is defined as agreement among 95% or more of results from all participants or referees. AAB PROFICIENCY TESTING SERVICE ~ 2008 Program Guide 17

20 2008 PROGRAM GUIDE Proficiency Testing Service AAB GRADING PROCEDURES continued SPECIAL GRADING FORMULAS IN MICROBIOLOGY When consensus grading is applied to microbiology modules in which more than one response may be reported for a given procedure in the same specimen, special formulas for generating percent scores for each specimen are mandated by the February 28 Rule of These formulas thus govern the grading of organism identifications and/or antimicrobial susceptibilities in Bacteriology, GC Culture, Parasitology, Throat, Throat/Urine and Urine Culture programs. For scoring organism identifications in a given specimen, the following ratio of responses is multiplied by 100: (1) The numerator is the sum of all gradable correct participant responses. Only one participant response per organism is counted, and, in cases in which more than one response is given per organism, only the most definitive answer is used. For example, a species name takes precedence over a genus name which, in turn, takes precedence over an antigen screen result which, in turn, takes precedence over a Gram stain result. (2) The denominator is the sum of all incorrect responses for the presence of organisms plus the total number of gradable correct responses for the specimen. Since AAB determines which organism(s) to place in each specimen, AAB determines the total number of gradable correct responses for the presence of organisms. For scoring antimicrobial susceptibilities in a given specimen, the following ratio of responses is multiplied by 100: (1) The numerator is the sum of all gradable correct responses. (2) The numerator is the sum of all gradable correct responses. Since each participant determines which antimicrobials to report on their patient specimens, each participant determines the total number of antimicrobial susceptibilities to report on PT specimens. The only reporting requirement is that the susceptibilities reported on a proficiency test specimen reflect the same list of drugs which would be tested by the laboratory on patient material in the same clinical context as that of the proficiency specimen. PEER GROUPS FOR QUANTITATIVE RESULTS To provide optimal peer grading, quantitative results are grouped initially by coupling reagent and instrument. After outlier removal, if the group shows poor precision, a second grouping by reagent alone is applied. If a third group is required, method principle or calibration type is employed. If no information is provided concerning reagent and instrument, or if the group is not statistically viable, results are graded by total population. This style of grading is used for Alcohol, Basic/Comprehensive Chemistry, Coagulation, Direct Bilirubin, Fertility Endocrinology, Fructosamine, Glycohemoglobin, Isoenzymes, Lipids, Special Chemistry, Therapeutic Drug Monitoring and Urine Microalbumin. Other quantitative modules are grouped initially by instrument or method, second by method principle and finally by total population. This scheme applies to Activated Clotting Time, Blood Gases, Centrifugal Hematology (QBC), C-Reactive Protein, Hematology, Immunoproteins, Urinalysis, Whole Blood Glucose, Whole Blood Prothrombin Time, Andrology and Embryology programs. REFERENCE METHOD VALUES Reference method values for Cholesterol (HDL, LDL and Total) and Triglycerides are published in our Participant Statistics for informational purposes only. They are not used in grading in any fashion. We gratefully acknowledge Atherotech, Inc. for providing these values. WOULD REFER Would Refer is a special response available for some interpretive programs such as Microbiology, Clinical Microscopy or Hematology with Manual Differential. It represents the situation where you would refer a non-routine result on to another entity such as a reference laboratory or pathologist for further interpretation or for a final result. You may only enter this response if this is your laboratory s standard procedure for handling patient samples with similar results. You may not enter Would Refer for all 5 specimens in these programs unless you are not performing any patient testing of the same type at the time of the proficiency testing event. If you do report Would Refer for all 5 specimens, you will be graded as having discontinued testing for the current testing event only. TABLE OF GRADING LIMITS INTRODUCTION "Fixed" grading limits use either a constant concentration (type C limits) or a constant percentage of the target value (type P limits) to define a range of acceptable performance above and below the target value. The target value for qualitative or semi-quantitative results is the most frequently reported (consensus) value(s) of all valid results. The target value for quantitative results is the mean of all results after removal of outliers. In contrast to fixed limits, limits based on standard deviations (SDs) can lead to different grading ranges for the same analyte, even when different peer groups have the same means. The limits required by CMS consist of all three types discussed above - fixed concentration (type C), fixed percentage (type P) and 3SD (type S): type C- plus/minus a fixed concentration from the mean; used in the absence of type P limits or when these limits exceed type P limits type P- the mean plus/minus a fixed percentage of the mean; used in the absence of type C limits or when these limits exceed type C limits type S- plus/minus three standard deviations (SDs) from the mean; mandated by CMS in the absence of type C and/or type P limits All three types of CMS grading limits are used. Not all analytes listed have CMS limits. Proficiency testing is not required by CMS or other agencies for analytes/procedures which have no CMS-mandated grading limits. Such analytes/procedures are identified with a pound sign (#) in the table on the next page as well as on the reporting forms. Results for these CMS-unscored analytes are not included in computing participants' overall CMS subspecialty scores. Therefore these scores do not appear on CMS Cumulative Summary reports; they appear only on graded reports (which do not go to CMS). In the interest of uniformity, the limits used for CMS-unscored analytes are derived from those used for CMS-scored analytes which are similar. A listing of the limits used for all analytes/procedures, both CMS-scored and CMS-unscored, follows Program Guide ~ PROFICIENCY TESTING SERVICE AAB