Lab O. Sharon L. Rolando, M.S., MT(ASCP)

BD Newsletter June 2001 Volume 12 No. 2 Indispensable to human health Lab O MICROBIOLOGY NEWS & IDEAS In This Issue... TechniTopic: Update on Pertussis and Pertussis-like Organisms 3 BACTEC System News: BD Donates BACTEC MGIT 960 Instruments to Russia New Blood Transfer Device for BACTEC Bottles Introduced BD BACTEC MGIT 960 AST for PZA Launched Outside the U.S. 5 Product Hi-Lights: BD BBL CHROMagar Orientation Medium Introduced New BDProbeTec ET Studies Published BD BBL Streptocard Individual Latex Components Available BD Bactrol Plus Offerings Expanded Reintroducing - BD Cellmatics Viral Transport Pack 9 FYI: BD Difco - BBL Gram Stain Technical Update BBL Sensi-Disc QC Log Sheets Available on the Internet International Symbols and Their Meaning - What's New? 2000-2001 Flu Season Was Mild New QC/PI Manual Now Available Coming Soon - Certificates of Analysis On-Line Update on Pertussis and Pertussis-like Organisms Sharon L. Rolando, M.S., MT(ASCP) Members of the genus Bordetella cause contagious acute respiratory disease in humans and other animals. B. pertussis, the organism most commonly associated with the human disease whooping cough, continues to cause outbreaks despite the widespread use of an effective childhood vaccine. Other species of Bordetella are also associated with human disease, generally causing milder respiratory symptoms. Classic whooping cough is characterized by three distinct stages of illness. 1 Initial symptoms are non-specific, resembling a cold or other viral illness, and may last 1-2 weeks. Following this initial catarrhal stage, cough becomes predominant. The patient may exhibit the characteristic whoop sound on inspiration, and a series of coughs may be followed by vomiting. This stage, known as the paroxysmal stage, may last 2-6 weeks. The final convalescent stage may persist for many weeks and involves coughing episodes of reduced severity. Whooping cough is most contagious during the 2-week catarrhal stage, passing from person-to-person via respiratory droplets. Frequently, however, diagnosis is not made until the paroxysmal stage, when the characteristic cough occurs. Disease outbreaks frequently occur, involving unvaccinated children or previously vaccinated adolescents and/or adults in whom immunity has waned. In older patients, the symptoms may be mild and atypical, making correct diagnosis and treatment difficult. Causative Agents B. pertussis is the most commonly recovered species of Bordetella. It is a fastidious gram-negative coccobacillus that requires special growth media for cultivation. The organisms attach to the cilia of respiratory epithelial cells and produce toxins that cause inflammation and interfere with host immune responses. The major virulence factor of B. pertussis is the production of pertussis toxin, Bordetella pertussis, the causative agent of whooping cough. although many other factors are now thought to play significant roles in the disease process. In infants, pneumonia or serious neurological complications can be associated with infection, including death on rare occasions. While most whooping cough infections are caused by B. pertussis, B. parapertussis is known to cause a milder form of the disease. This decreased pathogenicity of B. parapertussis is likely due to a lack of expression of the pertussis toxin and the absence of other virulence factors. Additional Bordetella species

TECHNITOPIC have also been isolated from patients with pertussis-like symptoms, including B. bronchiseptica (the cause of kennel cough in dogs) and B. holmesii (formerly CDC non-oxidizer group 2). The epidemiology and pathogenicity of these latter two species have not been completely studied. In fact, B. holmesii has only recently been isolated from human respiratory tract specimens. The previous failure to detect B. holmesii is most likely due to the presence of inhibitory antibiotics in the media commonly used for specimen transport. 2 Laboratory Diagnosis Laboratory testing requires the collection of a nasopharyngeal (NP) swab or aspirate for direct antigen detection, culture and/or polymerase chain reaction (PCR). NP aspirates are preferred, as they are suitable for all testing methodologies and can be stored in Casamino Acid broth for several days. If an aspirate collection is not clinically indicated, NP swabs may be collected. Calcium alginate swabs are best suited for culture, while polyester swabs are preferred for PCR testing. When the swab specimen cannot be plated immediately at the bedside, a transport medium must be used. Casamino Acid broth may be used if the transport time is expected to be <2 hours. Longer transport times require either Amies medium with char- Sharon L. Rolando, M.S., MT(ASCP) Sharon Rolando received her bachelor's degree in medical technology from Marquette University in 1992 and her master's degree in molecular microbiology and immunology from The Johns Hopkins School of Hygiene and Public Health in 1998. Her master's thesis, published by The Johns Hopkins School of Public Health, was titled "The Emergence of Human Infection with Penicillium marneffei in Relation to the Treatment and Control Patients with confirmed cases of B. pertussis should be put in respiratory isolacoal (<24 hours transport) or Regan- Lowe agar (>24 hours transport). The most rapid method for pertussis diagnosis involves microscopic examination of the collected material. The specimen is fixed to a glass slide and stained with polyclonal fluorescent antibody reagents against B. pertussis and B. parapertussis. This test can be completed within a few hours, but requires highly trained technologists to distinguish true positive results from the non-specific staining that is seen with some organisms. 3 The sensitivity of this method is poor, and its specificity is hampered by cross-reactivity with Bordetella bronchiseptica. Dual testing of all patient specimens with both FA reagents can help detect such cross-reactivity. The gold standard for diagnosis of Bordetella infection remains culture of the causative agent. All species of Bordetella may be identified by their biochemical reactions, and organisms suspected of being B. pertussis or B. parapertussis may be confirmed with direct fluorescent antibody stains. HIV Epidemic in Southeast Asia." She has worked as a clinical microbiologist for nine years, including seven years at The Johns Hopkins Hospital where she specialized in infectious disease serology. Currently, Ms. Rolando is completing work as an Emerging Infectious Diseases Fellow through the Centers for Disease Control and Prevention/ National Center for Infectious Diseases and Association of Public Health Laboratories at the Massachusetts Department of Public Health in Boston, Mass. Her areas of interest include the study of the molecular epidemiology of Neisseria gonorrhoeae and multi-drug resistant organisms. Regan-Lowe is an enriched medium for the selective recovery of B. pertussis. Regan-Lowe Charcoal Agar can also be used as a transport medium. Culture is highly specific, but sensitivity is frequently limited by inadequate specimen collection and transport procedures. The turn-around-time for culture may be as long as 14 days, and definitive culture identification of B. holmesii requires a transformation assay to rule-out the closely related Acinetobacter species. Assays that utilize PCR provide rapid, sensitive, and specific results, and they do not require that viable organisms be present in the sample. However, expensive and sophisticated equipment is required, and strict quality control measures must be implemented. Additionally, standardization between laboratories has generally not been achieved. Finally, PCR assays that target the IS481 repetitive element of B. pertussis have been shown to cross-react with B. holmesii. 4 Serological diagnosis of whooping cough is not widely available. The IgG assay is suitable only for individuals older than 11 years of age, as IgG titers in younger children would indicate vaccine status rather than a current infection. Although measurable IgA titers rarely occur in response to vaccination, they are only present in 40-60% of infected patients, and therefore are not suitable for diagnostic testing. 5 Neither IgG nor IgA antibodies are detectable in serum until approximately 14 days after the onset of symptoms. Thus, no single assay is completely adequate for the laboratory diagnosis of pertussis-like disease. All test results must be interpreted together and with the addition of clinical or epidemiological data. 2 Lab O volume 12 number 2 Continued on page 11

BACTEC SYSTEM NEWS BD Donates BACTEC MGIT 960 Instruments to Russia On March 20, 2001, BD symbolically donated two BACTEC MGIT 960 instruments to Russia during a ceremony at the Russian Embassy in Berlin, Germany. The ceremony was attended by representatives of the German government and the Russian and American embassies, as well as leaders from the scientific community. The BD BACTEC MGIT 960 instruments, along with some supplies, are being donated to two medical institutes in Moscow the Central Institute of Tuberculosis of the Russian Academy of Medical Sciences and the Moscow Center of Scientific and Practical Fight against Tuberculosis. It is expected that this donation will help in establishing a state-of-the art culture and drug susceptibility test system for tuberculosis (TB) and will offer an improvement in the diagnosis of multi-drug resistant tuberculosis (MDR-TB). In addition, these instruments will be used in a collaborative study to develop new tools for TB Some of the ceremony participants in front of the Russian Embassy in Berlin, Germany (from left to right): Dr. Larry Warfel (BD, U.S.), Dr. Gerhard J. Müller (LMTB, Germany), Dr. Olga V. Demikhova (Russian Federation), Mr. Detlef S. Siewert (BD, Eastern Europe), Dr. Salman H. Siddiqi (BD, U.S.), Dr. Klaus W. Berndt (BD, U.S.), Dr. Jürgen W. Leonhardt (I.U.T., Germany) and Mr. Hans-Joachim Cappius (LMTB, Germany). diagnosis such as an Electronic Super Nose an instrument designed to noninvasively detect volatile target molecules of tuberculosis. In this study, the BACTEC MGIT 960 instruments will serve as the reference method to determine the sensitivity and specificity of the Electronic Super Nose. If successful, the Electronic Super Nose could one day be used to rapidly screen people for tuberculosis. The study is being organized by scientists from BD, two German companies (the Laser and Medical Technology Center Berlin [LMTB] and the Institute for Environmental Technologies [ I.U.T.]) and the Russian institutes. Tuberculosis is an increasingly serious health problem in Russia. There are over three million cases of TB and in some prisons, as many as 80% of the inmates are infected. 1 Even more alarming is the high incidence of MDR-TB in Russia, estimated to be 37%. 1 It is hoped that the results of this study will lead to methods for earlier diagnosis of this rapidly spreading disease. For a complete overview of the worldwide TB crisis, see the LabO article Tuberculosis- A Public Health Crisis in the February 2001 issue, available at http://www.bd.com/clinical. 1 Halter et al. Der Spiegel. Jan. 22, 2001. Lab O volume 12 number 2 3

BACTEC SYSTEM NEWS BD Introduces New Blood Transfer Device for BACTEC Bottles The new BD Blood Transfer Device is a one-piece device designed to safely transfer blood from standard syringes into BACTEC blood culture bottles or evacuated blood collection tubes. This new device protects the health and safety of nurses, phlebotomists and others who draw blood by reducing the risk of spills and needlesticks during the blood transfer process. At the same time, it helps to ensure blood sample integrity, which is critical for accurate diagnosis and prescription. Each sterile, individually wrapped BD Blood Transfer Device consists of a plastic adapter fitted with a Luer-Lok tip and back-end needle; i.e., the needle is inside the adapter. After blood is drawn by syringe, the Blood Transfer Device is attached to the syringe via the Luer-Lok tip and the entire unit is used to fill the desired blood collection tubes and/or bottles. Multiple tubes/bottles may be filled sequentially from a single draw. If an indwelling catheter is used to initially draw the blood, there are no exposed needles from start to finish and the risk of needlestick injuries is minimized. For more information on the BD Blood Transfer Device, Catalog No. 364880, visit our electronic catalog at http://catalog.bd.com. BD BACTEC MGIT 960 Antimicrobial Susceptibility Test for PZA* Launched Outside the U.S. Pyrazinamide (PZA) susceptibility testing on the BD BACTEC MGIT 960 System is now available outside the U.S. The BACTEC MGIT 960 PZA susceptibility test augments existing BACTEC MGIT 960 SIRE* testing, which provides a susceptible or resistant result for four other drugs commonly used to treat tuberculosis streptomycin, isoniazid, rifampin and ethambutol. Multi-drug resistant Mycobacterium tuberculosis (MDR-TB) has recently become a serious public health problem. 1 Resistance to any of the primary drugs, including pyrazinamide, makes the disease more difficult and expensive to treat. Therefore, the rapid detection of these strains is critical to the effective treatment of the patient. The BACTEC MGIT 960 PZA Medium is a tube containing a modified Middlebrook 7H9 Broth that supports the growth and detection of mycobacteria at a ph of 5.9. The MGIT tube contains a fluorescent compound embedded in silicone on the bottom of a 16 x 100 mm round-bottom tube. The fluorescent compound is sensitive to the presence of oxygen dissolved in the broth. The initial concentration of dissolved oxygen quenches the fluorescent emission from the compound and little fluorescence can be detected. Later, actively respiring microorganisms consume the oxygen, which allows the compound to fluoresce. The BACTEC MGIT 960 PZA Kit is a qualitative test that is completed in 4 to 21 days by monitoring the growth of M. tuberculosis in a drug-containing tube compared to a drug-free tube (Growth Control). The BACTEC MGIT 960 instrument continually monitors tubes for increased fluorescence, indicating the presence of active microorganisms. Susceptible or resistant results are produced when the instrument compares the drug-containing tube to the control tube. The BACTEC MGIT 960 PZA Kit has been developed to allow susceptibility testing at a PZA concentration of 100 µg/ml. This concentration correlates with the concentration used in the BACTEC 460TB System. The latter is the method recommended by the National Committee for Clinical Laboratory Standards (NCCLS) for PZA susceptibility testing. *Not available in the U.S. 1 Barenfanger, J. 1993. Clin. Microbiol. Newsl. 15:76. 4 Lab O volume 12 number 2

PRODUCT HI-LIGHTS Now Available BBL CHROMagar Orientation Medium BD Diagnostic Systems is pleased to announce the introduction of its second chromogenic prepared plated medium BBL CHROMagar Orientation Medium for the detection and presumptive identification of urinary tract pathogens. BBL CHROMagar Orientation Medium joins BBL CHROMagar Candida Medium, in BD's chromogenic media portfolio. BBL CHROMagar Orientation Medium is a non-selective medium for the direct identification, differentiation and enumeration of grampositive and gram-negative pathogens associated with urinary tract infections (UTI). Differences in colony color and morphology facilitate detection of mixed cultures in specimens. The medium contains artificial substrates (chromogens) that release differently colored compounds upon degradation by specific microbial enzymes. The respective organisms cultured on the media react in such a way that the colonies take on a characteristic color and colonial morphology. This enables the direct differentiation of certain species or the detection of certain groups of organisms with only a minimum of confirmatory tests. Escherichia coli, Enterococcus, the Klebsiella-Enterobacter-Serratia (KES) and the Proteus-Morganella-Providencia (PMP) groups are frequently encountered organisms in UTIs. Most UTIs are caused by E. coli alone, or in combination with enterococci. Staphylococcus saprophyticus and Streptococcus agalactiae may be isolated from females, although less frequently. The respective organisms... take on a characteristic color and colonial morphology. is superior to other differential media used in the isolation, differentiation and enumeration of UTI pathogens, such as CLED Agar or a combination of Blood and MacConkey agars. 1-3 BBL CHROMagar Orientation Medium allows the differentiation of E. coli, enterococci, and most strains of S. saprophyticus directly on the primary isolation plate. Presumptive identification of Klebsiella-Enterobacter-Serratia and the Proteus-Morganella-Providencia groups is possible by means of colony morphology, pigmentation and medium discoloration. As this medium is nonselective, other UTI pathogens will grow, but will not show characteristic pigmentation, requiring additional biochemical tests to confirm identification. For more information on BBL CHROMagar Orientation Medium, mark the appropriate box on the reader response card. 1 Merlino et al. 1996. J. Clin. Microbiol. 34:1788. 2 Hengstler, Hammann and A.-M. Fahr. 1997. J. Clin. Microbiol. 35:2773. 3 Forbes, Sahm and Weissfeld (ed.). 1998. Bailey & Scott's diagnostic microbiology, 10th ed. Mosby, Inc., St. Louis. E. coli (rose) and Enterobacter cloacae (blue) Proteus sp. (brown) and Staphylococcus epidermidis (white) Enterococcus sp. (blue) and Staphylococcus epidermidis (white) Clinical studies have demonstrated that BBL CHROMagar Orientation Medium E. coli (rose) and Enterococcus sp. (blue) Lab O volume 12 number 2 5

PRODUCT HI-LIGHTS New BDProbeTec ET Studies Published Multicenter Evaluation of the BDProbeTec ET System for Detection of Chlamydia trachomatis and Neisseria gonorrhoeae in Urine Specimens, Female Endocervical Swabs, and Male Urethral Swabs Journal of Clinical Microbiology, Vol. 39, No.3, March 2001 The BDProbeTec ET System for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) was evaluated by seven centers. Swab and urine specimens were collected from 2109 men and women, with or without symptoms, attending sexually transmitted disease, family planning and obstetrics /gynecology clinics (4131 specimens). Specimens were tested by the BDProbeTec ET test, the Abbott LCx test (Abbott Laboratories) and culture. The performance of the BDProbeTec ET and Abbott systems versus patient infection status is summarized below. CT BDProbeTec ET Abbott LCx Sensitivity Specificity Sensitivity Specificity Female Swabs 92.8% 98.1% 94.4% 99.1% Female Urine 80.5% 98.4% 77.2% 99.0% Male Swabs 92.5% 96.4% ND ND Male Urine 93.1% 93.8% 93.8% 96.2% ND = Not Done GC BDProbeTec ET Abbott LCx Sensitivity Specificity Sensitivity Specificity Female Swabs 96.6% 99.5% 92.1% 99.8% Female Urine 84.9% 99.4% 83.7% 99.4% Male Swabs 98.5% 96.5% ND ND The investigators concluded Male Urine 97.9% 96.5% 93.8% 96.1% that the BDProbeTec ET System was comparable in ND = Not Done performance to the Abbott LCx System. In addition, they agreed that the BDProbeTec ET System "was easy to use, required low maintenance and had high throughput. The closed-system design and assay workflow were seen as advantageous." 6 Lab O volume 12 number 2

PRODUCT HI-LIGHTS Performance Characteristics of the Becton Dickinson ProbeTec System for Direct Detection of Chlamydia trachomatis and Neisseria gonorrhoeae in Male and Female Urine Specimens in Comparison with the Roche COBAS System Archives of Pathology and Laboratory Medicine, Vol. 124, Nov. 2000 Chan and colleagues from the Provincial Laboratory in Saskatchewan, Canada evaluated the performance of the BDProbeTec ET System versus the Roche COBAS AMPLICOR System (Roche Diagnostic Systems) for the detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) in urine specimens. A total of 1224 urine specimens, 825 male and 399 female, were tested by both assays. An in-house confirmatory PCR test was performed on discrepant specimens. Amplification controls were included in all three assays to monitor inhibition. After resolution, performance was as follows: CT Sensitivity Specificity PPV NPV BDProbeTec ET System 95.3% 99.3% 95.9% 99.2% Roche COBAS System 95.9% 98.3% 90.6% 99.3% GC Sensitivity Specificity PPV NPV While the sensitivities of both systems were comparable, the Roche sys- BDProbeTec ET System 100% 99.7 % 88.2% 100% Roche COBAS System 96.7% 98.9% 69% 99.9% tem was less specific with 30 false positive results compared to 11 for the BDProbeTec ET system. In fact, the Roche system produced a PPV of 69% for GC. The investigators commented that cross-reactivity with nongonococcal Neisseria species has been previously observed in the Roche system. The inhibition rates for this study were 3.3% and 2.0% for the BDProbeTec ET and Roche systems, respectively. When these specimens were retested, all produced negative results in both systems. The investigators also evaluated both systems for throughput. They found that the BDProbeTec ET system was capable of testing 150 specimens for CT and GC using an amplification control in an 8-hour shift using one instrument. The maximum throughput for the Roche system was 46 specimens using one instrument per 8- hour shift. The investigators concluded the high throughput and performance of the BDProbeTec ET instrument allow us to achieve increased sensitivity and specificity with greater efficiency than previously possible. For a copy of either of these articles or for more information on the BDProbeTec ET System, mark the appropriate box(es) on the reader response card. Lab O volume 12 number 2 7

PRODUCT HI-LIGHTS BD BBL Streptocard C, F and G Individual Latex Components Now Available Individual components for the C, F and G latex reagents of the BD BBL Streptocard Acid Latex Test Kit and the BD BBL Streptocard Enzyme Latex Test Kit are now available, completing the individual component offerings for these kits. The BBL Streptocard Acid Latex Test Kit is a test system for the qualitative identification of β-hemolytic colonies of Lancefield streptococcal groups A, B, C, F and G, based on specific carbohydrate antigens. The BBL Streptocard Enzyme Latex Test Kit is a test system for qualitative identification of β-hemolytic and nonhemolytic colonies of Lancefield streptococcal groups A, B, C, D, F and G, based on specific carbohydrate antigens. The individual kit components (50 tests each) are: Catalog No. Description 240960 Strep Group A Latex 240961 Strep Group B Latex 240865 Strep Group C Latex 240962 Strep Group D Latex 240866 Strep Group F Latex 240867 Strep Group G Latex 240963 Acid Extraction Reagent 240964 Enzyme Extraction Reagent 240965 Positive Control 240966 Reaction Cards For more information on these products, visit our website at http://www.bd.com/- clinical/products/stains/strepg.html. BD Bactrol Plus Offerings are Expanding The BD Bactrol Plus line of Quality Control Cultures is expanding with the addition of 11 new organisms. With these new cultures, the BD Bactrol Plus offerings now total 29 cultures and 5 sets. The BD Bactrol Plus Quality Control Cultures were introduced last year and will be replacing the well-known Bactrol disks. The Bactrol Plus cultures are vials of lypholized microorganisms. As such, they are easily reconstituted and there is less chance of contamination compared to the handling requirements of disks. After reconstitution, the resulting suspension is ready for inoculation onto appropriate plating media. The cultures are used to assist in the quality control of microbiological media, reagents and identification systems. The organisms are derived strains that are maintained by nationally recognized culture collections, such as the American Type Culture Collection (ATCC ). These organisms have consistent biochemical characteristics or known susceptibility patterns and can be used as internal controls to evaluate an individual laboratory's procedures and practices. BD Bactrol Plus vials are for single use and come packaged in pouches of 5 vials. For more information, check the appropriate box on the reader response card. BD Cellmatics Viral Transport Pack the Complete System for Quality Viral Collection and Transport When it comes to assuring quality viral test results, proper specimen collection and transport are two of the most critical components of the diagnostic process. The quality and accuracy of all test results from the laboratory 8 Lab O volume 12 number 2 are contingent on the quality of the specimens received. The BD Cellmatics Viral Transport Pack offers a convenient, high-performance system to meet the demanding viral collection and transport needs of your laboratory. Each BD Cellmatics Viral Transport Pack includes: One screw-cap vial containing 2 ml of liquid, tissue-culture compatible viral transport medium. Difco brand viral transport medium contains Hanks balanced salt solution with gelatin as a holding medium, HEPES biological buffer as used in cell culture media and gentamicin sulfate to reduce the overgrowth of microorganisms. One sterile, rayon-tipped, plastic shaft swab. The rayon-tipped swab preserves the integrity of viruses without causing cell culture toxicity problems. One transport pouch the cardboard transport pouch with printed instructions and patient ID label, protects sample during transport. For more information on the BD Cellmatics Viral Transport Pack, contact your local BD sales representative.

FYI BD Difco - BBL Gram Stain Technical Update We are pleased to announce that BD Diagnostic Systems has developed and incorporated a unique filtration process improvement to reduce the presence of artifacts when using BD Difco-BBL Gram Stains. Why these improvements? Safranin counterstain contains varying amounts of insoluble, spherical, chemical precipitate that could be mistaken for gram-positive cocci in Gram stain preparations. These spherical artifacts are normally 0.5 µm or smaller in diameter and appear dark purple under 1000X bright field microscopy. The spheres are present in low numbers, from 0 to 10 per 20 fields examined, depending upon the source of the powdered stain and the methods used to prepare the stain. The spheres are usually single, rarely in pairs, but never appear in clusters or chains. How to distinguish artifacts? Artifacts can be differentiated from gram-positive coccoidal bacteria by careful examination of the Gram stain preparation. Bacteria that have been properly fixed on the microscope slide during the staining process appear in a single plane. Artifacts, on the other hand, sit on top of the plane of fixed bacteria and can be brought into sharp focus by slightly raising the microscope objective with the fine adjustment knob. They are most easily distinguished from bacterial cells by examination of the Gram stained slide under 1000X phase contrast microscopy; bacteria appear pinkish, pale blue or opaque white whereas artifacts appear as refractile, bright spheres. With the implementation of the filtration step in the manufacturing process for safranin counterstain, the frequency of artifacts has been significantly reduced resulting in Gram stain preparations that are clearer and easier to read. At BD, we are committed to producing the highest quality products to meet your laboratory needs. For more information on BD Difco -BBL Gram Stains, mark the appropriate box on the reader response card. BD BBL Sensi-Disc Quality Control Log Sheets Available on the Internet To assist you with your antimicrobial disc susceptibility testing documentation, we have produced up-to-date Quality Control Log Sheets for BD BBL Sensi-Disc Antimicrobial Discs available on the Internet. The log sheets contain the 86 Sensi-Disc antimicrobial agents recommended by the National Committee for Clinical Laboratory Standards (NCCLS), along with their corresponding quality control organism zone sizes, and space to record disc lot numbers and test results. The log sheets, two 8.5" x 14.5" or 8.5" x 11" sheets, may be downloaded and photocopied for repeated use. Visit our website at http://www.bd.com/- clinical/products/idsus/sensi.html and download your copy of the BD BBL Sensi-Disc Quality Control Log Sheets today. An alphabetized list of the 86 Sensi-Disc antimicrobial agents. Space to record disc lot numbers. Expected organism zone sizes and space to record observed zone sizes. Lab O volume 12 number 2 9

FYI International Symbols and Their Meaning What's New? As BD Diagnostics Systems products expand into the international marketplace, the task of communicating information to a multi-national, multi-lingual customer base becomes increasingly challenging. One method to meet this challenge is to utilize internationally recognized symbols that communicate specific information without words or the need for translation. BD Diagnostic Systems has committed to the use of these symbols and has been actively converting product labeling to include their use. The use of these symbols provides many benefits to both BD and the customer: Fewer words on the labels which reduces "clutter" Easily recognized graphics that provide immediate information In the September 1999 edition of LabO, we provided a glossary of the symbols that are used on our product labeling. In addition, many of our products are packaged with a copy of the symbol glossary for your use. With this and future editions of LabO, we will be providing explanations of individual symbols to highlight their use and assist in familiarizing you with their meaning. For a copy of the symbol glossary, mark the appropriate box on the reader response card or visit our website at http://www.bd.com/clinical/index.html. Lot Number Symbol Uniformity in labeling format on each product label The actual lot number is presented to the side of this symbol. Expiration Date Symbol The actual expiration date is presented to the side of this symbol. With the change in the millennium and new international requirements, the format of the expiration date has been revised. Both the old and new formats are shown to the right. The new format will also be used on Certificates of Analysis in the near future. Old Formats (Expiration Date of January 15, 2002) DD-MMM-YY Example: 15 JAN 02 YY/MM/DD Example: 02/01/15 New Format (Expiration Date of January 15, 2002) YYYY/MM/DD or YYYY/MM a four (4) digit year a two (2) digit month a two (2) digit day (if necessary) Example: 2002/01/15 or 2002/01 10 Lab O volume 12 number 2

FYI 2000-2001 Flu Season Was Mild The Centers for Disease Control and Prevention (CDC) surveillance report for the week ending April 7, indicates that the current flu season was milder than the past three seasons. This year's peak positivity rate was 24% (occurring at the end of January) compared to peak positivity rates of 28% to 33% for the last three seasons. Although influenza A viruses have predominated this year, influenza type B viruses have been more prevalent than in previous years. Since October 1, World Health Organization and National Respiratory and Enteric Virus Surveillance System laboratories have identified 9,325 positive influenza specimens of which 55% were influenza type A (primarily H1N1) and 45% were influenza type B. In fact, influenza type B viruses have predominated (range 53% to 62%) in the Mountain, Pacific and Mid- Atlantic regions. The CDC provides influenza surveillance data from October through May on its website http://www.cdc.gov/ncidod/diseases/flu/- weekly.htm. Information is updated weekly. Update on Pertussis and Pertussis-like Organisms Continued from page 2 tion, kept from contact with nonimmunized infants, and treated with appropriate antibiotic therapy. 6 Although antibiotic therapy may not lessen the duration of symptoms, it will decrease the period of communicability. Additionally, all contacts of a known positive patient, regardless of immunization status, should be sought for prophylactic antibiotic treatment. 6 Erythromycin is the antibiotic of choice for treating cases of B. pertussis and their contacts. 5 Trimethoprim-sulfamethoxazole (TMP-SMX) is also effective, whereas amoxicillin and oral cephalosporins are not effective. Susceptibility testing of Bordetella species has not been standardized. Methods that have been utilized for research purposes include agar dilution, broth dilution, gradient strip and disc diffusion. Gradient strip results have been shown to correlate well with agar dilution results for several drugs, including erythromycin. 7 Erythromycin resistant isolates of B. pertussis are extremely rare, even in widespread surveillance testing. 8 Childhood vaccination is the best control measure for prevention of whooping cough. While the efficacy of both the whole-cell and the new acellular vaccine has been proven, the acellular preparation is more commonly used, as there are fewer complications. Childhood vaccination is the best control measure for prevention of whooping cough. Vaccination, including booster shots, is contraindicated for individuals older than 7 years of age due to severe adverse reactions, especially with the whole cell preparation. There is evidence that neither the whole-cell nor the acellular pertussis vaccine provides protection against species of Bordetella other than B. pertussis. 9 In conclusion, as long as B. pertussis remains an endemic infection in the United States, accurate diagnosis and reporting will remain the core of effective control measures. Research into new methods to detect this organism will continue to improve the laboratory's ability to aid clinicians in making this important diagnosis. The role of other species of Bordetella in causing human disease needs to be defined, as well as the mechanisms of resistance that may be developing in this vaccinepreventable disease. 1 Koneman et al. (ed.). 1997. Color atlas and textbook of diagnostic microbiology, 5th ed. Lippincott-Raven Publishers, Philadelphia. 2 Mazengia et al. 2000. J. Clin. Microbiol. 38:2330. 3 Becton Dickinson. Bacto FA Bordetella Pertussis/ Parapertussis package insert. Becton Dickinson, Sparks. 4 Loeffelholz et al. 2000. J. Clin. Microbiol. 38:467. 5 Hoppe. 1999. Bordetella. In Murray, Baron, Pfaller, Tenover and Yolken (ed.), Manual of clinical microbiology, 7th ed. ASM, Washington D.C. 6 Chin (ed.). 2000. Pertussis. In Control of communicable diseases manual, 17th ed. APHA, Washington, D.C. 7 Hill et al. 2000. J. Clin. Microbiol. 38:1151. 8 Korgenski et al. 1997. J. Clin. Microbiol. 35:2989. 9 Mastrantonio et al. 1998. J. Clin. Microbiol. 36:999. LabO is published three times per year by Becton, Dickinson and Company, 7 Loveton Circle, Sparks, MD 21152-0999, 410.316.4701. Editor: Mary Jo Zimbro, B.S., MT(ASCP). Send address changes and mailing list additions to the attention of Marketing Communications, Mail Code 634. For technical information, call Technical Services, toll free, at 800.638.8663. Visit our website at http:www.bd.com/clinical CHROMagar is a trademark of Dr. A. Rambach. Bacto, Bactrol, Cellmatics and Difco are trademarks of Difco Laboratories, subsidiary of Becton, Dickinson and Company. BD, BD Logo and all other trademarks are the property of Becton, Dickinson and Company unless otherwise noted. 2001BD BD Diagnostic Systems is an ISO 9000 registered manufacturer. Becton, Dickinson and Company 2001 BD Printed in USA Lab O volume 12 number 2 11

FYI Recently Released The New Edition of the BBL Quality Control and Product Information Manual The new edition of the BBL Quality Control and Product Information (QC/PI) Manual was released in April. The newly updated Manual has been condensed and contains product information for both plated and tubed media (previously, the information was contained in separate plate and tube manuals). Thus, the new manual replaces the previous versions of both the Plate and Tube Manuals. The Manual was distributed to all prepared media customers on our QC/PI mailing list. Customers were sent either a printed hard copy or a CD- ROM of the new manual. The new edition of the QC/PI Manual includes: Changes in quality control procedures and test organisms made by our Quality Control Laboratory Updated organism names to adopt recent changes in microbial taxonomy Revised text and references, where appropriate, to provide information on current microbiological procedures and identification tables Two new plated media, Selective Streptococcus Agar and MacConkey II Agar with Sorbitol Users of BBL plated and tubed media who do not have these manuals or who did not receive the April mailing, should visit our website http://www.bd.com/clinical or call BD Diagnostic Systems Technical Services at 800-638-8663, prompt 2, to order a copy. And don't forget to advise us if the person designated to receive and maintain these manuals changes by returning the postage-paid business reply card located at the back of each manual. Coming Soon Certificates of Analysis On-Line Certificates On Line is a new system that provides Internet access to Certificates of Analysis for specific microbiology products from BD Diagnostic Systems. Certificates of Analysis will be available on the Internet 24 hours a day, 7 days a week this summer. Customers worldwide will simply register on the BD website and enter product catalog numbers and lot numbers to immediately receive the desired Certificates of Analysis. This new service is free-ofcharge, fully automated and also allows for batch requests; i.e., customers may request multiple Certificates at the same time. After the system retrieves the requested Certificates, customers have the option of e-mailing them to other associates, printing or downloading the files. Animal origin information will also be included on the Certificates of Analysis for applicable products. Certificates of Analysis and Certificates of Origin are routinely requested for the purposes of incoming product testing, FDA inspections and documentation in QC files, among others. In these scenarios, Certificates are often needed immediately. Providing access to these Certificates on the Internet enables customers to conveniently access these documents without delays in turnaround time. Access to Material Safety Data Sheets (MSDS) and package inserts will be added in the future. Certificates On Line augments the existing system of Certificates On Call (formerly called Bactofax), whereby customers also have 24-hour access to Certificates by phone and fax machine. Certificates On Call will continue to be available to customers and can be reached in the U.S. by calling 800-343- 2655. For requests originating outside the U.S., calls should be made to 410-773-6926. Additionally, customers may continue to contact BD Technical Services at 800-638-8663, selection 2, for further information. To receive notification of Certificates On Line going live, register at http://www.bd.com/diagnostics/regslive.html. We will also send you a complimentary brochure describing the new system. BD Diagnostic Systems 7 Loveton Circle Sparks, MD 21152 800.638.8663 www.bd.com/diagnostics