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1 ABBOTT PRISM HCV Hepatitis C Virus Encoded Antigens (Recombinant c100-3, HCr43, NS5) en ABBOTT PRISM HCV 6A /R10 B6A520 Read Highlighted Changes Revised March, 2008 Customer Service For additional product information, please contact your local customer service organization. NOTE: This package insert must be read carefully prior to product use. Package insert instructions must be carefully followed. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in this package insert. NOTE: If you receive reagents, calibrators, controls or bulk solutions that are in a condition contrary to the package insert or label recommendation, or that are damaged, contact your local customer service organization. For use with software version 2.1 or higher Key to symbols used List Number In Vitro Diagnostic Medical Device Lot Number Expiration Date Store at 2-8 C Store at C CAUTION: Consult accompanying documents. Manufacturer Master Lot Sample Cups Calibrators Pipette Tips Line Cleaner Assay Kit Card Reaction Trays Reagent Components Run Control Adapters See REAGENTS section for a full explanation of symbols used in reagent component naming. ABBOTT Diagnostics Division 1

2 U. S. Patent No. 4, 380, 580, licensed under patent rights of Bayer Corp. Tarrytown, New York, USA, relates to this product. NAME AND INTENDED USE The ABBOTT PRISM HCV assay is an in vitro chemiluminescent immunoassay (ChLIA) for the qualitative detection of antibodies to hepatitis C virus (anti-hcv) in human serum or plasma. The ABBOTT PRISM HCV ChLIA is intended as a screen for donated blood to prevent transmission of hepatitis C virus (HCV) to recipients of blood and blood components and as an aid in the diagnosis of hepatitis C viral infection. SUMMARY AND EXPLANATION OF THE TEST Serological studies to detect the antibodies to recombinant antigens of HCV have established HCV as the cause of most blood-borne, 1-6 as well as community acquired, 7 non-a, non-b hepatitis (NANBH). Thus, the presence of anti-hcv indicates that an individual may have been infected with HCV, may harbor infectious HCV and may be capable of transmitting HCV infection. 8 However, as with all immunoassays, the ABBOTT PRISM HCV assay may yield non-specific reactivity due to other causes, particularly when testing low prevalence populations. Although the majority of infected individuals may be asymptomatic, complications of HCV infection may include chronic hepatitis, cirrhosis and increased risk of hepatocellular carcinoma The implementation of blood donor screening for anti-hcv has led to a marked decline in the risk of transfusion-transmitted hepatitis. 13,14 The ABBOTT PRISM HCV is designed to detect antibodies to recombinant antigens covering Core, NS3, NS4, and NS5 regions. The relationship between the recombinant proteins used for the test and the putative structural and nonstructural proteins of the HCV genome 15 is depicted in the figure below. Serological studies of HCV infection indicate that antibodies may recognize any or all of the regions of the HCV genome represented on the ABBOTT PRISM HCV solid phase, thereby improving the sensitivity of anti-hcv detection The HCr43 protein, expressed in Escherichia coli (E. coli), is composed of two non-contiguous coding regions of the Chiron HCV genome sequence. The first of the two regions represents amino acids #1192 to 1457 of the Chiron HCV sequence known as clone 33. The second region represents amino acids #1 to 150 of the Chiron HCV sequence. The genomic organization of other flaviviruses suggests the cloned sequence is the core coding sequence of HCV. This construct is therefore referred to as 33c-Core by Abbott. The c100-3 protein, expressed in Saccharomyces cerevisiae (S. cerevisiae), includes amino acids #1569 to 1931 of the Chiron HCV genome sequence. The genomic organization of other flaviviruses suggests the cloned sequence is contained within the putative non-structural (NS3 and NS4) regions of HCV. The NS5 protein, expressed in S. cerevisiae, includes amino acids #2054 to 2995 of the Chiron HCV genome sequence. The genomic organization of other flaviviruses suggests the cloned sequence is contained within the putative non-structural region of HCV referred to as NS5. HCV antigens c100-3, NS5 and HCr43 are prepared by Chiron Corporation under a shared manufacturing agreement. BIOLOGICAL PRINCIPLES OF THE PROCEDURE The ABBOTT PRISM HCV assay is a two-step sandwich ChLIA that detects antibodies to hepatitis C virus in human serum or plasma. The reactions occur in the following sequence: Microparticles coated with HCV recombinant antigens are incubated with Specimen Diluent and either plasma, serum, calibrator or control in the incubation well of the reaction tray. During incubation, HCV antibodies present in the sample bind to the antigen(s) on the microparticles. After this first incubation is complete, the reaction mixture is transferred to the glass fiber matrix (matrix) of the reaction tray using the Transfer Wash. The microparticles are captured by the matrix while the remaining mixture flows through to the absorbent blotter. The anti-biotin (mouse monoclonal): acridinium conjugate/biotinylated F(ab ) 2 fragment (goat) anti-human IgG is added to the microparticles on the matrix and incubated. After this second incubation, the unbound conjugate is washed into the blotter with the Conjugate Wash. The chemiluminescent signal is generated by addition of an alkaline hydrogen peroxide solution. The resultant photons are counted. The amount of light emitted is proportional to the amount of antibody to HCV in the sample. For further information regarding ChLIA technology, refer to the ABBOTT PRISM Operations Manual, Section 3. The presence or absence of antibody to HCV in the sample is determined by comparing the number of photons collected from the sample to a cutoff value determined from an ABBOTT PRISM calibration performed in the same batch. If the number of photons collected from a test sample is greater than or equal to the cutoff value, the sample is considered reactive for antibody to HCV. Specimens which are not reactive by the ABBOTT PRISM HCV assay are considered nonreactive for antibody to HCV. These specimens need not be further tested. Specimens which are initially reactive should be centrifuged according to the table in the SPECIMEN COLLECTION AND PREPARATION FOR ANALYSIS section and retested in duplicate. REAGENTS NOTE: Each specific component description noted below is accompanied by a unique symbol. These symbols appear on both the component labels and on corresponding instrument tubing identifier labels. They are meant to facilitate identification and installation of reagent bottles within the ambient reagent bay and refrigerator. ABBOTT PRISM HCV Assay Kit (6A52-48) 1 bottle (325 ml) hepatitis C virus encoded antigens (recombinant c100-3, HCr43, NS5) coated microparticles in phosphate buffered saline. Preservative: sodium azide. (Symbol: ) 1 bottle (332 ml) anti-biotin (mouse monoclonal): acridinium conjugate/biotinylated F(ab ) 2 fragment (goat) anti-human IgG (gamma) in phosphate buffer, bovine serum albumin and Triton X-100. Preservative: sodium azide. (Symbol: ) 2

3 3 bottles ( 8.3 ml each) Negative Calibrator. Recalcified human plasma nonreactive for HBsAg, HIV-1 RNA or HIV-1 Ag, Anti-HCV and Anti-HIV-1/HIV-2. Preservative: sodium azide. (Symbol: NC) 3 bottles ( 8.3 ml each) Positive Calibrator. Recalcified, heat-inactivated human plasma reactive for Anti-HCV, nonreactive for HBsAg and Anti-HIV-1/HIV-2, HIV-1 RNA or HIV-1 Ag. Preservative: sodium azide. (Symbol: PC) 1 bottle (328 ml) Specimen Diluent. Borate buffered saline with Tween 20, bovine serum albumin, and Triton X-100. Preservative: sodium azide. (Symbol: X) ABBOTT PRISM HCV Wash Kit (6A52-38) 1 bottle (3360 ml) Transfer Wash. Borate buffered saline with Tween 20. Preservative: sodium azide. (Symbol: ~) 1 bottle (1734 ml) Conjugate Wash. MES {2-(N-morpholino) ethanesulfonic acid} buffered saline. Preservative: ProClin 300. (Symbol: ) Other Reagents Required ABBOTT PRISM Activator Concentrate (1A75-02 or 3L27-02) 4 bottles (900 ml each) Activator Concentrate. 0.4% hydrogen peroxide/0.06% diethylenetriaminepentaacetic acid. ABBOTT PRISM Activator Diluent (1A75-01 or 3L27-01) 4 bottles (900 ml each) Activator Diluent. 0.3 N sodium hydroxide. Other Reagents Available ABBOTT PRISM Run Control Kit (5E22-10) ABBOTT PRISM Positive Run Control Kit (5E22-11) ABBOTT PRISM Run Control Kit (2K24-10) ABBOTT PRISM Positive Run Control Kit (2K24-11) NOTE: Each batch MUST end in a release control. An HCV release control is any control reactive for anti-hcv which has been configured to validate system function and release sample results. The configuration criteria are defined in the RUN CONTROLS file of the ABBOTT PRISM Resource Management software. Any customer specified control reactive for anti-hcv may be used. WARNINGS AND PRECAUTIONS For In Vitro Diagnostic Use. CAUTION: This product contains human sourced infectious and/or potentially infectious components. Refer to the REAGENTS section of this package insert. No known test method can offer complete assurance that products derived from human sources or inactivated microorganisms will not transmit infection. Therefore, it is recommended that all human sourced materials be considered potentially infectious and handled with appropriate biosafety practices. Safety Precautions The ABBOTT PRISM Line Cleaner containing 2% tetraethylammonium hydroxide (TEAH) may cause mild eye irritation. If this solution comes in contact with eyes, rinse immediately with water (for additional information, refer to the ABBOTT PRISM Operations Manual, Section 8). Conjugate Wash contains methylisothiazolones which are components of ProClin and is classified per applicable European Community (EC) Directives as: Irritant (Xi). The following are the appropriate Risk (R) and Safety (S) phrases. R43 May cause sensitization by skin contact. S24 Avoid contact with skin. S35 This material and its container must be disposed of in a safe way. S37 Wear suitable gloves. S46 If swallowed, seek medical advice immediately and show this container or label. The ABBOTT PRISM Activator Diluent contains sodium hydroxide and is classified per the applicable European Community (EC) Directives as: Irritant (Xi). The following are the appropriate Risk (R) and Safety (S) phrases. R36 Irritating to eyes. S26 In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. S35 This material and its container must be disposed of in a safe way. S46 If swallowed, seek medical advice immediately and show this container or label. This product contains sodium azide; for a specific listing, refer to the REAGENTS section. Contact with acids liberates very toxic gas. This material and its container must be disposed of in a safe way. For product not classified as dangerous per European Directive 1999/45/EC as amended - Safety data sheet available for professional user on request. Handling Precautions CAUTION: If the ABBOTT PRISM HCV assay is selected, put on clean gloves before handling HCV conjugate. HCV conjugate is neutralized by contamination with human IgG. Extreme caution must be exercised when handling all containers, tubing, and accessories which may come in contact with the conjugate. Do not use kits beyond the expiration date. Gently invert each component several times prior to loading on the ABBOTT PRISM System to ensure a homogenous solution. Avoid foaming. Each component of the ABBOTT PRISM HCV Wash Kit should be at room temperature (15 to 30 C) before mixing. Do not mix reagents from different bottles. Do not mix reagents from different assay kit lots. Any lot of ABBOTT PRISM HCV Wash Kit can be used with any lot of ABBOTT PRISM HCV Assay Kit. Avoid microbial and chemical contamination of samples, reagents, and equipment. The use of disposable pipette tips is recommended for any preliminary sample transfer. Do not freeze reagents. Failure to adhere to instructions in the ABBOTT PRISM Operations Manual or Package Insert may result in erroneous results. Use caution when handling samples, reagent bottles, and reagent caps to prevent cross contamination. Additional safety and handling precautions and limitations for the assay kit, calibrators, specimens, controls, and other reagents are described in the ABBOTT PRISM Operations Manual, Section 7. PREPARATION OF ACTIVATOR SOLUTION Activator Solution is prepared daily by mixing equal parts of Activator Concentrate and Activator Diluent. The volume of Activator Solution required for multiple tests is calculated by the ABBOTT PRISM software. Refer to the ABBOTT PRISM Operations Manual, Section 5, under Plan Work Load, for additional information. Use clean pipettes and/or metal-free containers (such as plasticware or acid-washed and distilled or deionized water-rinsed glassware) to measure. Prepare in the bottle provided in the Accessory Kit. Cover the bottle opening securely with the cap provided and invert gently five to ten times to mix. Load the Activator Solution on the PRISM System. Refer to the ABBOTT PRISM Operations Manual, Section 5, under Prepare and Load Activator Solution, for additional information. NOTE: The Activator Solution must be used within 24 hours of preparation. 3

4 STORAGE INSTRUCTIONS Store the ABBOTT PRISM HCV Assay Kit, ABBOTT PRISM Run Control Kit, ABBOTT PRISM Positive Run Control Kit, and ABBOTT PRISM Activator Concentrate at 2 to 8 C. Store the ABBOTT PRISM HCV Wash Kit and ABBOTT PRISM Activator Diluent at room temperature (15 to 30 C). Store ABBOTT PRISM Pipette Tips and ABBOTT PRISM Reaction Trays in their original package until use. INDICATIONS OF INSTABILITY OR DETERIORATION OF REAGENTS The ABBOTT PRISM System will not continue to process samples when calibrator values do not meet specifications. This may indicate either deterioration or contamination of reagents, or instrument failure. Refer to the ABBOTT PRISM Operations Manual, Section 10, for additional information. INSTRUMENT PROCEDURE Refer to the ABBOTT PRISM Operations Manual for a detailed description of Instrument Procedures. Solutions required for instrument cleaning and maintenance are described in detail in the ABBOTT PRISM Operations Manual, Sections 5 and 9. For optimal performance, it is important to follow the routine maintenance procedures defined in the ABBOTT PRISM Operations Manual, Section 9. SPECIMEN COLLECTION AND PREPARATION FOR ANALYSIS Either serum (including serum collected in serum separator tubes) or plasma collected in EDTA, Potassium Oxalate, Sodium Citrate, ACD, CP2D, CPD, or CPDA-1 anticoagulants may be used with the ABBOTT PRISM HCV assay. NOTE: Do not use specimens collected in heparin. Use of heparin as an anticoagulant may cause a reduction in Sample Net Counts and in S/CO (Sample Net Counts/Cutoff). Therefore, heparin is not recommended for any PRISM Assay. This assay was designed and validated for use with human serum or plasma from individual donor specimens. Pooled specimens must not be used. Heat-inactivated specimens should be avoided. Gravity separation is not sufficient for specimen preparation. Specimens collected by plasmapheresis which have not been frozen do not require centrifugation. All other specimens (including previously frozen plasmapheresis specimens) must be centrifuged. Non-frozen specimens (excluding non-frozen plasmapheresis specimens) must be centrifuged such that g-minutes (the product of relative centrifugal force [RCF] and centrifugation time [minutes]) is between 30,000 and 75,000. The following chart lists acceptable time and force ranges that meet this criteria. RCF x Time Time (minutes) RCF (x g) (g-minutes) 10 3,000 30, ,000-3,000 30,000-45, ,500-3,000 30,000-60, ,300-3,000 32,500-75,000 Convert rpm to RCF as follows: RCF = 1.12 r max (rpm/1000) 2 Convert RCF to rpm as follows: rpm = 1000 x RCF 1.12 x r max RCF - The relative centrifugal force generated during centrifugation. rpm - The rotation per minute of the rotor on which the specimens are being spun (usually the digital readout on the centrifuge will indicate rpm). Time - The time should be measured from the time the rotor reaches the required RCF or rpm to the time it begins decelerating. r max - The radius of the rotor in millimeters. The radius measured is dependent on whether the rotor is a fixed angle rotor or a swinging bucket rotor. This value is typically provided with the rotor, by the manufacturer. For fixed angle, r max is a measure of the distance from the rotor axis (center) to the bottom of the tube cavity. For the swinging bucket, r max is a measure of the distance from the rotor axis (center) to the bottom of the tube bucket while it is extended during rotation. g-minutes - The unit of measure for the product of RCF (x g) and time (minutes). Previously frozen specimens must be centrifuged such that g-minutes (the product of relative centrifugal force [RCF] and centrifugation time [minutes]) is between 180,000 and 300,000. The following chart lists acceptable time and force ranges that meet this criteria. RCF x Time Time (minutes) RCF (x g) (g-minutes) 15 12, , ,000-12, , , ,200-12, , ,000 ANY specimen (excluding non-frozen plasmapheresis) not tested within 24 hours of initial centrifugation, must be recentrifuged from 30,000 to 75,000 (g-minutes) as defined for non-frozen specimens. NOTE: If re-testing a specimen within 24 hours of initial centrifugation, the specimen is not required to be re-centrifuged. FAILURE TO FOLLOW THE SPECIFIED CENTRIFUGATION PROCEDURE MAY GIVE ERRONEOUS OR INCONSISTENT RESULTS. Specimens may be stored at 2 to 8 C for up to fourteen days. If storage periods greater than fourteen days are anticipated, the serum or plasma should be removed from the clot or red blood cells to avoid hemolysis. Store the serum or plasma frozen (-10 C or colder). Previously frozen specimens must be mixed thoroughly after thawing and centrifuged according to the table in this section. 10 nonreactive and 10 low-level reactive specimens showed no qualitative performance differences when subjected to 6 freeze-thaw cycles. However, specimens that have undergone multiple freeze-thaw cycles or have been stored frozen for prolonged periods may give erroneous or inconsistent results. Clear, non-hemolyzed specimens should be used when possible. Specimens containing particulate matter may give erroneous or inconsistent test results. No qualitative performance differences were observed when nonreactive and low-level reactive specimens were spiked with elevated levels of bilirubin ( 20 mg/dl), hemoglobin ( 500 mg/dl), or lipids ( 3000 mg/dl). When shipped, specimens must be packaged and labeled in compliance with applicable regulations covering the transport of clinical specimens and etiologic agents. Specimens may be shipped under ambient conditions, refrigerated on wet ice (2 to 8 C), or frozen on dry ice (-10 C or colder). Prior to freezing, the specimen should be removed from the clot or red cells. Performance has not been established for cadaver specimens, umbilical cord blood, or body fluids such as urine, saliva, semen, amniotic fluid, cerebrospinal fluid, or pleural fluid. 4

5 Specimen Volume The specimen volume required to perform a single assay on the ABBOTT PRISM System varies according to the different specimen containers. For ABBOTT PRISM Sample Cups, the minimum specimen volume required for one assay is 350 μl. The ABBOTT PRISM HCV Assay requires 50 μl sample dispense. For volume requirements for each additional assay performed from the same specimen container and for volume requirements in primary or aliquot tubes, refer to the ABBOTT PRISM Operations Manual, Section 5. ABBOTT PRISM HCV PROCEDURE Materials Provided 6A52-48 ABBOTT PRISM HCV Assay Kit* 6A52-38 ABBOTT PRISM HCV Wash Kit Materials Required but not Provided 1A75-02 or 3L27-02 ABBOTT PRISM 1A75-01 or 3L27-01 ABBOTT PRISM 5A07-01 ABBOTT PRISM 5A07-10 ABBOTT PRISM 6A36-60 ABBOTT PRISM Accessory Kit Additional Materials Available 1A75-10 or 3L27-10 ABBOTT PRISM 2K24-10 ABBOTT PRISM Run Control Kit 2K24-11 ABBOTT PRISM Positive Run Control Kit 5E22-10 ABBOTT PRISM Run Control Kit 5E22-11 ABBOTT PRISM Positive Run Control Kit 6A36-31 ABBOTT PRISM 7A03-01 or 3L00-01 ABBOTT PRISM 7A03-30 or 3L00-30 ABBOTT PRISM 7A03-31 ABBOTT PRISM 7B36-01 ABBOTT PRISM ABBOTT PRISM ASSAY PROCEDURE For detailed information concerning batch time and maximum batch size, refer to the ABBOTT PRISM Operations Manual, Section 2. STEP 1: Enter a Plan Work Load (refer to ABBOTT PRISM Operations Manual, Section 5). Replace reagents as needed. NOTE: Gently invert each component several times prior to loading on the ABBOTT PRISM System to ensure a homogenous solution. Additional gentle inversion may be required to thoroughly resuspend microparticles. Avoid foaming. Each component of the ABBOTT PRISM HCV Wash Kit should be at room temperature (15 to 30 C) before mixing. Verify that all tubing label symbols match the symbols on each reagent label. (Refer to the symbol key in the REAGENTS section.) Verify that all tubing is securely fastened to the corresponding wash and reagent bottles. STEP 2: Inspect the waste containers. Empty and clean as defined in the ABBOTT PRISM Operations Manual, Section 9, if necessary. STEP 3: Prepare Activator Solution (refer to the PREPARATION OF ACTIVATOR SOLUTION section of this package insert) and load into the ABBOTT PRISM System. STEP 4: Verify adequate number of Reaction Trays are in the Tray Loader. STEP 5: Verify adequate number of Pipette Tips are in the Pipette Tip Racks. STEP 6: Perform the prime procedure (refer to the ABBOTT PRISM Operations Manual, Section 5). STEP 7: Initiate sample processing. Open the bottles in the Calibrator Pack and place in the Calibrator Rack. Load the Calibrator Rack and Sample Racks, including the Controls (refer to the Control Handling Procedure, under Controls). STEP 8: After the calibrators have been pipetted, remove Calibrator Rack. Close the Calibrator bottles and return to 2 to 8 C storage. STEP 9: Each sample is initially tested once. Sample Racks may be removed after the samples have been pipetted. STEP 10: Any specimen (excluding non-frozen plasmapheresis) that is initially reactive must be centrifuged according to the table in the SPECIMEN COLLECTION AND PREPARATION FOR ANALYSIS section and retested in duplicate (refer to the ABBOTT PRISM Operations Manual, Section 5). NOTE: Specimens retested within 24 hours of initial centrifugation do not require recentrifugation. STEP 11: After the run is complete, perform the purge procedure (refer to the ABBOTT PRISM Operations Manual, Section 5). Refer to the ABBOTT PRISM Operations Manual, Section 3, for a detailed description of ChLIA procedures. QUALITY CONTROL PROCEDURES Calibration The ABBOTT PRISM HCV Negative and Positive Calibrators are tested in triplicate automatically at the beginning of each batch. The ABBOTT PRISM System will not release results when Calibrator values do not meet specifications. This may indicate either deterioration or contamination of reagents, or instrument failure. Controls 1. A release control MUST be included as the last sample in each batch. A control such as the ABBOTT PRISM Positive Control or any customer-specified control reactive for anti-hcv may be used. This control must test reactive in order to validate system functionality and to release results. If this control does not test reactive, refer to the ABBOTT PRISM Operations Manual, Section 10. Sites using a release control other than the ABBOTT PRISM Positive Control must validate its performance on the ABBOTT PRISM System. 2. ABBOTT PRISM Run Control Handling Procedure a. Place each Run Control bottle into an adapter such that when the flip-top cap is opened, it can be snapped into an open position within the adapter. b. Place each Run Control within an adapter onto the Sample Rack. The controls can be placed in any rack position except 1, 2, 27, or 28. c. As mentioned above, place an ABBOTT PRISM Positive Control after the last sample tested in the batch. The controls can be placed in any rack position except 1, 2, 27, or 28. * Average expected kit utilization is 4100 tests per kit. Actual utilization will vary by customer. 5

6 3. Customer-Specified Control Handling Procedure a. Determine the volume of controls required. The control volume required to perform a single assay on the ABBOTT PRISM System varies according to the different specimen containers. For ABBOTT PRISM Sample Cups, the minimum control volume required for one assay is 500 μl (300 μl μl Sample Cup dead volume). For every additional assay performed from the same control container, an additional 100 μl is required. For volume requirements in primary or aliquot tubes, refer to the ABBOTT PRISM Operations Manual, Section 5. b. Refer to the ABBOTT PRISM Operations Manual, Section 5: Operating Instructions, Subsection: Sample Processing. 4. Additional controls may be run at the operator s discretion. Validity specifications may be assigned such that if these controls fail, no results are reported for that assay batch. ASSAY PARAMETER SPECIFICATIONS The PRISM assay parameter specifications have been factory set. These parameters cannot be printed, displayed, or edited. RESULTS Calculation of Cutoff and S/CO Values The ABBOTT PRISM System calculates the ABBOTT PRISM HCV assay cutoff value using the following formula: Cutoff = Mean Negative Calibrator Net Counts + (0.55 x Mean Positive Calibrator Net Counts)* Example: If the Mean NCC = 2,500, and the Mean PCC = 30,000, 2,500 + (0.55 x 30,000) = 19,000 Cutoff = 19,000 * The Mean Positive Calibrator and Negative Calibrator Net Counts are calculated using the two lowest replicates. An instrument code (02-211) will be displayed in place of the count value for the third replicate. Each of the two remaining replicates of the Positive Calibrator and the Negative Calibrator used to calculate the cutoff must meet all specifications. The ABBOTT PRISM System calculates the ABBOTT PRISM HCV assay S/CO using the following formula: S/CO = Sample Net Counts / Cutoff Example: If the Sample Net Counts = 32,000, and the Cutoff = 19,000, 32,000 / 19,000 = 1.68 S/CO = 1.68 INTERPRETATION OF RESULTS In the ABBOTT PRISM HCV assay, specimens with Net Counts less than the cutoff are considered nonreactive and need not be further tested. Specimens with Net Counts greater than or equal to the cutoff are considered initially reactive. All specimens that are reactive on initial testing must be centrifuged according to the table in the SPECIMEN COLLECTION AND PREPARATION FOR ANALYSIS section of this package insert and retested in duplicate. NOTE: If re-testing a specimen within 24 hours of the initial centrifugation, the specimen is not required to be re-centrifuged. If repeat testing shows the Net Counts for both retests to be less than the cutoff, the sample is considered nonreactive. If either duplicate retest Net Count is greater than or equal to the cutoff, the specimen is repeatedly reactive. Repeatedly reactive specimens should be investigated further in supplemental tests such as other HCV specific immunoassays and immunoblot assays or a combination thereof. ABBOTT PRISM reports display sample results in Net Counts and/or S/CO. Net Counts are used by ABBOTT PRISM to interpret results. The S/CO value is provided in reports to show relative reactivity to the cutoff. In the ABBOTT PRISM HCV assay, specimens with S/CO values of less than 1.00 are considered nonreactive. Specimens with an S/CO value of greater than or equal to 1.00 are considered reactive. For details on configuring the ABBOTT PRISM System to use Grayzone Interpretations, refer to the ABBOTT PRISM Operations Manual, Section 2. READING RESULTS Some S/CO values may be flagged with < or > symbols. Refer to the ABBOTT PRISM Operations Manual, Section 5: Operating Instructions, Subsection: Reports. SYSTEM ERRORS For a description of the error codes that appear in the ABBOTT PRISM Report, refer to the ABBOTT PRISM Operations Manual, Section 10. LIMITATIONS OF THE PROCEDURE Testing of previously frozen samples with the ABBOTT PRISM HCV assay may cause erroneous results. This assay was designed and validated for use with human serum or plasma from individual patient and donor specimens. Pooled specimens must not be used since the accuracy of their test results has not been validated. It is recommended that repeatedly reactive specimens be investigated by supplemental testing. Individuals who are repeatedly reactive may be referred for medical evaluation which may include additional testing. False reactive results can be expected with any test kit. Falsely elevated results have been observed due to non-specific interactions (see Table II). Performance has not been established for body fluids other than serum or plasma. Previously frozen specimens must be centrifuged per the SPECIMEN COLLECTION AND PREPARATION FOR ANALYSIS section of this package insert prior to running the assay. Serum from heparinized patients may be incompletely coagulated. Erroneous or inconsistent results may occur due to the presence of fibrin. To prevent this phenomenon, draw specimen prior to heparin therapy. Do not use specimens collected in heparin. Use of heparin as an anticoagulant may cause a reduction in Sample Net Counts and in S/CO (Sample Net Counts/Cutoff). The use of specimens with obvious microbial contamination should be avoided. Although the association of infectivity and the presence of anti-hcv is strong, it is recognized that presently available methods for anti-hcv detection are not sensitive enough to detect all potentially infectious units of blood or possible cases of HCV infection. EXPECTED RESULTS In a random population of 6,742 volunteer blood donor specimens, 26 (0.39%) were reactive by ABBOTT PRISM HCV. Among 419 specimens from individuals with acute or chronic HCV infection and preselected anti-hcv positive specimens, the ABBOTT PRISM HCV assay detected % as repeatedly reactive. SPECIFIC PERFORMANCE CHARACTERISTICS NOTE: Representative performance data are shown. Results obtained at individual laboratories may vary. Precision Assay reproducibility was determined by assaying a 16 member panel consisting of four replicates each of three diluted specimens reactive for anti-hcv (panel members 1, 2, and 3) and one specimen nonreactive for anti-hcv (panel member 4). The panel was tested in five runs over five days with each of three master lots at a total of three sites. The intra- and inter-assay standard deviation (SD) and percent coefficient of variation (%CV) were analyzed with a variance components analysis, using a nested analysis of variance model 19 (Table I). Mean S/CO is defined as the mean Sample Net Counts (NET) divided by the calculated Cutoff. 6

7 TABLE I ABBOTT PRISM HCV Reproducibility Number of Mean Intra-assay Inter-assay a Panel Member Replicates S/CO SD %CV SD %CV Negative Control Positive Control Number of Mean Intra-assay Inter-assay a Calibrators Replicates NET SD %CV SD %CV Negative 180 3, Positive ,476 2, , Specificity The specificity of the ABBOTT PRISM HCV assay was estimated assuming a zero prevalence of HCV in volunteer blood donors and plasmapheresis donors. A total of 6,742 serum and plasma specimens from volunteer blood donors and plasmapheresis donors was collected from four blood centers (Table II). Of the 26 repeatedly reactive specimens eight were excluded as confirmed positive by ABBOTT MATRIX HCV 2.0 and/or RIBA 2.0. b Therefore, of the 6,734 donations presumed seronegative for anti-hcv, ABBOTT PRISM HCV had an estimated specificity of 99.73% (6,716/6,734). Specimens from individuals with medical conditions unrelated to HCV infection or containing potentially interfering substances and specimens from random hospital patients were tested with the ABBOTT PRISM HCV assay (Table II). TABLE II Reactivity in Donor Populations, in Random Hospital Patients, and in Specimens from Individuals with Medical Conditions Unrelated to HCV Infection or Containing Potentially Interfering Substances ABBOTT PRISM HCV Number of PRISM HCV Repeatedly Reactives Number Initially Repeatedly that were of Specimens Reactive Reactive Confirmed Group/Type n (%) n (%) Positive c Tested (%) Volunteer Serum 3, (0.39) 12 (0.39) 2 (16.67) Blood Donors Plasma 3, (0.44) 12 (0.38) 6 (50.00) Plasmapheresis (0.40) 2 (0.40) 0 (0.00) Donors TOTAL DONORS 6, (0.42) 26 (0.39) 8 (30.77) Random Hospital (7.59) 23 (7.59) 18 (78.26) Patients Medical Conditions Unrelated to HCV Infection and Potentially Interfering Substances d (9.80) 30 (9.80) 30 (100.00) Detectability Specimens obtained from patients with acute and chronic HCV infections, preselected anti-hcv positive specimens, and populations at increased risk of HCV infection were tested with the ABBOTT PRISM HCV assay. The ABBOTT PRISM HCV assay detected all of the 496 confirmed positive specimens (100.00%) (Table III). TABLE III Reactivity of the ABBOTT PRISM HCV Assay in Selected Populations with HCV Infection, or at Increased Risk of HCV Infection Number Number Number Repeatedly Confirmed Positive that of Reactive Number are ABBOTT Specimens by ABBOTT Con- PRISM HCV Group Tested PRISM HCV (%) firmed Positive Repeatedly Reactive (%) Selected Acute (100.00) 8 8 (100.00) Populations with HCV Infection Chronic (100.00) (100.00) Preselected Anti-HCV (100.00) (100.00) Positive Populations Populations at (49.40) (100.00) Increased Risk of HCV Infection e TOTAL (85.64) (100.00) Sensitivity The sensitivity of ABBOTT PRISM HCV was evaluated by testing 20 seroconversion panels and comparing to the ABBOTT HCV EIA 3.0. The ABBOTT PRISM HCV assay showed earlier detection than the ABBOTT HCV EIA 3.0 assay in eight of the 20 panels; five by detecting one additional bleed, two by detecting two additional bleeds, and one by detecting three additional bleeds. The ABBOTT PRISM HCV assay showed equivalent detection in ten of the 20 panels. The ABBOTT HCV EIA 3.0 showed earlier detection by one bleed in the remaining two panels. a) This term includes intra-assay variability. b) A confirmed positive result was defined as reactivity to 2 HCV gene products by ABBOTT MATRIX HCV 2.0 and/or RIBA 2.0. c) A confirmed positive result in these studies was defined as reactivity to 2 HCV gene products by ABBOTT MATRIX HCV 2.0 and/or RIBA 2.0. d) Medical Conditions Unrelated to HCV Infection and Potentially Interfering Substances included the following categories of frozen specimens: ANA Ab Positive, Rheumatoid Factor Positive, SLE, CMV Ab Positive, EBV Ab Positive, HAV Ab Positive, HBsAg Positive, HBc Ab Positive, HBs Ab Positive, HIV-1 Ab Positive, HSV Ab Positive, Rubella Ab Positive, Non-Viral Liver Disease, Multiple Myeloma, Syphilis Positive, Toxoplasma Ab Positive, Fungal Infections, Animal Handlers, Elevated Immunoglobulin, Elevated Bilirubin, Elevated Triglycerides, Elevated Cholesterol, Elevated Hemoglobin, Goat/Mouse Ab Positive, Multiparous Females, Multiple Transfusion Recipients, Vaccine Recipients, E. coli Infections, and Biotin Positive. e) Selected Populations at Increased Risk of HCV Infection included the following categories: U.S. IV Drug Users, Hemodialysis Patients, Homosexual Males, and Hemophiliacs. 7

8 BIBLIOGRAPHY 1. Choo Q.-L., Kuo G., Weiner A.J., Bradley D.W., and Houghton M. Isolation of a cdna Clone Derived From a Blood-Borne Non-A, Non-B Viral Hepatitis Genome. Science, 244: , Kuo G., Choo Q.-L., Alter H.J., et al. An Assay for Circulating Antibodies to a Major Etiologic Virus of Human Non-A, Non-B Hepatitis. Science, 244: , Alter H.J., Purcell R.H., Shih J.W., et al. Detection of Antibody to Hepatitis C Virus in Prospectively Followed Transfusion Recipients With Acute and Chronic Non-A, Non-B Hepatitis. N. Engl. J. Med., 321: , Esteban J.I., Viladomiu L., Gonzalez A., et al. Hepatitis C Virus Antibodies Among Risk Groups in Spain. Lancet, ii: , Van der Poel C.L., Lelie P.N., Choo Q.-L., et al. Anti-Hepatitis C Antibodies and Non-A, Non-B Post-Transfusion Hepatitis in the Netherlands. Lancet, ii: , Esteban J.I., Gonzalez A., Hernandez J.M., et al. Evaluation of Antibodies to Hepatitis C Virus in a Study of Transfusion Associated Hepatitis. N. Engl. J. Med., 323: , Miyamura T., Saito I., Katayama T., et al. Detection of Antibody Against Antigen Expressed by Molecularly Cloned Hepatitis C Virus cdna: Application to Diagnosis and Blood Screening for Posttransfusion Hepatitis. Proc. Natl. Acad. Sci. USA, 87: , Aach R.D., Stevens C.E., Hollinger F.B., et al. Hepatitis C Virus Infection in Post-Transfusion Hepatitis: An Analysis With First- and Second Generation Assays. N. Engl. J. Med., 325: , Alter M.J., Hadler S.C., Judson F.N., et al. Risk Factors for Acute Non-A, Non-B Hepatitis in the United States and Association With Hepatitis C Infection. JAMA, 264: , Choo Q.-L., Weiner A.J., Overby L.R., et al. Hepatitis C Virus: The Major Causative Agent of Viral Non-A, Non-B Hepatitis. Brit. Med. Bull., 46: , Dienstag J.L. Non-A, Non-B Hepatitis. I. Recognition, Epidemiology and Clinical Features. Gastro-enterology, 85: , Gitnick G. Non-A, Non-B Hepatitis: Etiology and Clinical Course. Ann. Rev. Med., 35: , Wick M.R., Moore S., Taswell H.F. Non-A, Non-B Hepatitis Associated With Blood Transfusion. Transfusion, 25:93-101, Hollinger F.B. Non-A, Non-B Hepatitis Viruses. In Virology, BN Fields and DN Knipe (Eds.). Raven Press, New York. pp , Japanese Red Cross Non-A, Non-B Hepatitis Research Group. Effect of Screening for Hepatitis C Virus Antibody and Hepatitis B Virus Core Antibody on Incidence of Post-Transfusion Hepatitis. Lancet, 338: , Donahue J.G., Alvaro Muno, D.V.M., A., Ness P.M., et al. The Declining Risk of Post-Transfusion Hepatitis C Virus Infection. N. Engl. J. Med., 327: , Choo Q.-L., Richman K.H., Han J.H., et al. Genetic Organization and Diversity of the Hepatitis C Virus. Proc. Natl. Acad. Sci. USA, 88: , McHutchinson J.G., Person J.L., Govindarajan S., et al. Improved Detection of Hepatitis C Virus Antibodies in High-Risk Populations. Hepatology, 15: 19-25, Statistics for Experiments: An Introduction to Design, Data Analysis, and Model Building. Box GEP, Hunter WG and Hunter JS. Wiley J and Sons, Inc., , All trademarks are property of their respective owners. ABBOTT Max-Planck-Ring Wiesbaden Germany March , 2008 Abbott Laboratories 8