ADVIA CENTAUR HBSAG READY PACK REAGENTS/CONFIRMATORY READY PACK REAGENTS/QUALITY CONTROL MATERIAL

P030049 · Siemens Healthcare Diagnostics · LOM · May 26, 2005 · Microbiology

Device Facts

Record IDP030049
Device NameADVIA CENTAUR HBSAG READY PACK REAGENTS/CONFIRMATORY READY PACK REAGENTS/QUALITY CONTROL MATERIAL
ApplicantSiemens Healthcare Diagnostics
Product CodeLOM · Microbiology
Decision DateMay 26, 2005
DecisionAPRL
Regulation21 CFR 866.3172
Device ClassClass 2
AttributesPediatric

Intended Use

The ADVIA Centaur® HBsAg assay is an in vitro immunoassay for the qualitative detection of hepatitis B surface antigen (HBsAg) in human serum and plasma (potassium EDTA, lithium or sodium heparinized,) using the ADVIA Centaur® system. The assay may be used in conjunction with other serological and clinical information to diagnose individuals with acute or chronic hepatitis B infection. The assay may also be used to screen for hepatitis B infection in pregnant women to identify neonates who are at risk of acquiring hepatitis B during the perinatal period. The ADVIA Centaur® HBsAg Confirmatory assay is an in vitro diagnostic immunoassay for the qualitative confirmation of the presence of hepatitis B surface antigen (HBsAg) in human serum or plasma (EDTA, lithium or sodium heparin,) using the ADVIA Centaur® system. The assay is intended to be used to confirm the presence of HBsAg in samples that are repeatedly reactive using the ADVIA Centaur® HBsAg assay.

Device Story

The ADVIA Centaur® HBsAg assay is a monoclonal antibody sandwich immunoassay using chemiluminometric technology. It processes human serum or plasma samples on the ADVIA Centaur® system. The assay uses biotinylated and acridinium ester-labeled anti-HBs monoclonal antibodies and streptavidin-coated magnetic latex particles. During incubation, HBsAg in the sample forms complexes with these antibodies and particles. After washing, acid and base are added to initiate a chemiluminescent reaction, measured as relative light units (RLUs) by a photomultiplier tube. The system calculates an Index Value from a stored Master Curve. The Confirmatory assay uses specific antibody neutralization; samples are pretreated with neutralizing (anti-HBs) or non-neutralizing reagents, and the inhibition of the RLU signal is compared to confirm HBsAg presence. Used in clinical laboratories by technicians to aid physician diagnosis of HBV infection and prenatal screening. Results assist clinicians in identifying infected individuals and managing neonates at risk of perinatal transmission.

Clinical Evidence

Clinical study evaluated 2,740 patients (high risk, signs/symptoms, dialysis, prenatal). HBV status classified via reference serological markers. Overall positive percent agreement was 94.57% (122/129) and negative percent agreement was 99.63% (1888/1895). Retrospective study of 153 acute/chronic HBV patients showed 100% positive agreement for acute and 98.1% for chronic. Prenatal study (n=716) showed 100% agreement. Seroconversion panels demonstrated detection timing comparable to reference methods. Precision/reproducibility studies showed acceptable variability across sites and lots.

Technological Characteristics

Monoclonal antibody sandwich immunoassay; chemiluminometric detection. Components: Streptavidin-coated magnetic latex particles, biotinylated/acridinium ester-labeled anti-HBs monoclonal antibodies. Energy source: Photomultiplier tube for RLU quantification. Connectivity: System-based, barcode-entered Master Curve. Sterilization: Not applicable (reagents). Software: Automated calculation of Index Values from stored calibration curves.

Indications for Use

Indicated for qualitative detection of HBsAg in human serum/plasma to diagnose acute/chronic HBV infection and for prenatal screening to identify neonates at risk of perinatal HBV acquisition. Confirmatory assay indicated for samples repeatedly reactive in the primary HBsAg assay.

Regulatory Classification

Identification

A qualitative hepatitis B virus (HBV) antigen assay is identified as an in vitro diagnostic device intended for prescription use for qualitative use with human serum, plasma, or other matrices that aids in the diagnosis of chronic or acute HBV infection. HBV surface antigen (HbsAg) is also used for screening of HBV infection in pregnant women to identify neonates who are at risk of acquiring hepatitis B during perinatal period. The assay is not intended for screening of blood, plasma, cells, or tissue donors.

Special Controls

*Classification.* Class II (special controls). The special controls for this device are:(1) The labeling required under § 809.10(b) of this chapter must include: (i) A prominent statement that the assay is not intended for the screening of blood, plasma, cells, or tissue donors. (ii) A detailed explanation of the principles of operation and procedures for performing the assay. (iii) A detailed explanation of the interpretation of results. (iv) Limitations, which must be updated to reflect current clinical practice and disease presentation and management. The limitations must include statements that indicate: (A) The specimen types for which the device has been cleared, and that use of this assay with specimen types other than those specifically cleared for this device may result in inaccurate assay results. (B) When appropriate, performance characteristics of the assay have not been established in populations of immunocompromised or immunosuppressed patients or other populations where assay performance may be affected. (C) Diagnosis of hepatitis B infection should not be established on the basis of a single assay result but should be determined by a licensed healthcare professional in conjunction with the clinical presentation, history, and other diagnostic procedures. (D) Detection of HBV antigens indicates a current infection with hepatitis B virus but does not differentiate between acute or chronic infection. False reactive HbsAg result may occur for up to 2 weeks after vaccination with HbsAg containing vaccine. (E) Current methods for the detection of hepatitis B antigens may not detect all potentially infected individuals. A non-reactive assay result does not exclude the possibility of exposure to or infection with hepatitis B virus. A non-reactive assay result in individuals with prior exposure to hepatitis B may be due to but not limited to antigen levels below the detection limit of this assay or lack of antigen reactivity to the antibodies in this assay. HBV mutants lacking the ability to produce antigens have been reported. These may occur as “escape” mutants in the presence of anti-HBV antibodies and such patients may be infectious. (F) Results obtained with this assay may not be used interchangeably with results obtained with a different manufacturer's assay. (2) Design verification and validation must include the following: (i) A detailed device description, including all parts that make up the device, ancillary reagents required but not provided, an explanation of the device methodology, design of the capture antibody(ies), external controls, and computational path from collected raw data to reported result ( *e.g.,* how collected raw signals are converted into a reported signal and result), as applicable to the detection method and device design.(ii) For devices with assay calibrators, the design and composition of all primary, secondary, and subsequent quantitation standards used for calibration as well as their traceability to a standardized reference material that FDA has determined is appropriate ( *e.g.,* a recognized consensus standard). In addition, analytical testing must be performed following the release of a new lot of the standard material that was used for device clearance or approval, or when there is a transition to a new calibration standard.(iii) Documentation and characterization ( *e.g.,* supplier, determination of identity, purity, and stability) of all critical reagents (including description of the capture antibody(ies)), and protocols for maintaining product integrity throughout its labeled shelf life.(iv) Risk analysis and management strategies, such as Failure Modes Effects Analysis and/or Hazard Analysis and Critical Control Points summaries and their impact on assay performance. (v) Final release criteria to be used for manufactured assay lots with appropriate evidence that lots released at the extremes of the specifications will meet the identified analytical and clinical performance characteristics as well as stability. (vi) Stability studies for reagents must include documentation of an assessment of real-time stability for multiple reagent lots using the indicated specimen types and must use acceptance criteria that ensure that analytical and clinical performance characteristics are met when stability is assigned based on the extremes of the acceptance range. (vii) All stability protocols, including acceptance criteria. (viii) Final release assay results for each lot used in clinical studies. (ix) Reproducibility study data that includes the testing of three independent production lots. (x) Detailed documentation of analytical performance studies conducted, as appropriate to the technology, specimen types tested, and intended use of the device, including, the limit of blank (LoB), limit of detection (LoD), cutoff, precision (reproducibility) including lot-to-lot and/or instrument-to-instrument precision, interference, cross reactivity, carryover, hook effect, seroconversion panel testing, matrix equivalency, prominent mutants/variants detection ( *e.g.,* for HbsAg), specimen stability, reagent stability, and cross-genotype antigen detection sensitivity, when appropriate.(xi) Analytical sensitivity of the assay that is the same or better than that of other cleared or approved assays. (xii) For devices with associated software or instrumentation, documentation must include a detailed description of device software, including software applications and hardware-based devices that incorporate software. The detailed description must include documentation of verification, validation, and hazard analysis and risk assessment activities, including an assessment of the impact of threats and vulnerabilities on device functionality and end users/patients as part of cybersecurity review. (xiii) Detailed documentation and results from a clinical study. Performance must be analyzed relative to an FDA cleared or approved HBV antigen assay or a comparator that FDA has determined is appropriate. This study must be conducted using appropriate patient samples, with an appropriate number of HBV reactive and non-reactive samples in applicable risk and disease categories, and any applicable confirmatory testing. Additional relevant patient groups must be validated as appropriate. The samples must include prospective (sequential) samples for each identified specimen type and, as appropriate, additional characterized clinical samples. Samples must be sourced from geographically diverse areas. This study must be conducted in the appropriate settings by the intended users to demonstrate clinical performance.

Related Devices

Submission Summary (Full Text)

{0} # Summary of Safety and Effectiveness ## I. GENERAL INFORMATION Device Generic Name: Hepatitis B Surface Antigen (HBsAg Assay) Hepatitis B Surface Antigen Confirmatory (HBsAg Conf assay) Hepatitis B Surface Antigen Positive and Negative Control materials Device Trade Name: ADVIA Centaur® HBsAg ReadyPack Reagents ADVIA Centaur® HBsAg Confirmatory ReadyPack Reagents ADVIA Centaur® HBsAg Quality Control Material Applicant’s Name and Address: Bayer HealthCare LLC 511 Benedict Avenue Tarrytown, NY 10591-5097 Date(s) of Panel Recommendation: None. Premarket Approval Application (PMA) Number: P030049 Date of Notice of Approval to Applicant: May 26, 2005 ## II. INDICATIONS FOR USE ADVIA Centaur® HBsAg Ready Pack Indications for use: The ADVIA Centaur® HBsAg assay is an *in vitro* immunoassay for the qualitative detection of hepatitis B surface antigen (HBsAg) in human serum and plasma (potassium EDTA, lithium or sodium heparinized,) using the ADVIA Centaur® system. The assay may be used in conjunction with other serological and clinical information to diagnose individuals with acute or chronic hepatitis B infection. The assay may also be used to screen for hepatitis B infection in pregnant women to identify neonates who are at risk of acquiring hepatitis B during the perinatal period. ADVIA Centaur® HBsAg Confirmatory Ready Pack Indications for use: The ADVIA Centaur® HBsAg Confirmatory assay is an *in vitro* diagnostic immunoassay for the qualitative confirmation of the presence of hepatitis B surface antigen (HBsAg) in human serum or plasma (EDTA, lithium or sodium heparin,) using the ADVIA Centaur® system. The assay is intended to be used to confirm the presence of HBsAg in samples that are repeatedly reactive using the ADVIA Centaur® HBsAg assay. ADVIA Centaur® HBsAg Quality Control Material Indications for Use: {1} Summary of Safety and Effectiveness For monitoring the performance of the HBsAg and HBsAg Confirmatory assays on the ADVIA Centaur® Systems. ## III. CONTRAINDICATIONS None known ## IV. WARNINGS AND PRECAUTIONS For in vitro diagnostic use only. Warnings and precautions for ADVIA Centaur® HBsAg ReadyPack Reagents, the ADVIA Centaur® HBsAg ReadyPack Calibrators, ADVIA Centaur® HBsAg Confirmatory ReadyPack Reagents, and the ADVIA Centaur® HBsAg Quality Control Material are stated in the respective product labeling. ## V. DEVICE DESCRIPTION ### Kit Configuration and Components The ADVIA Centaur® HBsAg assay contains the following components: | Reagent Pack | Reagent | Ingredients | | --- | --- | --- | | ADVIA Centaur® HBsAg ReadyPack primary reagent pack | Solid Phase | Streptavidin-coated magnetic latex particles in buffer with bovine serum albumin, goat serum, surfactant, sodium azide (<0.1%) and preservatives | | ADVIA Centaur® HBsAg ReadyPack ancillary reagent pack | Lite Reagent | Biotinylated monoclonal mouse anti-HBsAg antibody reactive to the “a” region determinant (~1.0 μg/mL) and acridinium ester-labeled monoclonal mouse anti-HBsAg antibody reactive to the “a” region determinant (~0.1 μg/mL) in buffer with bovine serum albumin, bovine gamma globulin, goat serum, mouse IgG, surfactant, sodium azide (<0.1%) and preservatives | | HBsAg calibrator vials | Calibrators | High calibrator: purified human HBsAg in buffer with sodium azide (<0.1%) low calibrator: processed normal human plasma with sodium azide (<0.1%) | The ADVIA Centaur® HBsAg Confirmatory assay contains the following components: - 1 ReadyPack primary reagent pack containing ADVIACentaur® HBsAg Solid Phase and Ancillary Reagent - ADVIACentaur® HBsAg Master Curve card - ADVIACentaur® HBsAg Confirmatory Master Curve card - 1 vial HBsAg Low Calibrator - 1 vial HBsAg High Calibrator - ADVIACentaur® HBsAg Calibrator Assigned Value card The ADVIA Centaur® HBsAg Controls contain the following components: - 2 vials of Negative Control - 2 vials of Positive Control - Expected Values Card and barcode 10 {2} Summary of Safety and Effectiveness The following are required, but not furnished with the ADVIA Centaur® HBsAg Assay: - ADVIA Centaur® Instrument - ADVIA Centaur® Ancillary Probe Wash 1 - ADVIACentaur® HBsAg quality control material - ADVIA Centaur® Wash 1 All human source components have been tested with FDA approved methods for anti-HIV 1 and 2, anti-HCV, HBV surface antigen and found to nonreactive. The human source calibrators and positive control contains HBsAg and are treated with a BPL-UV inactivation procedure to render the material noninfectious. ## Assay Principle and Procedure The ADVIA Centaur® HBsAg assay is a monoclonal antibody sandwich immunoassay using direct, chemiluminometric technology. Non-magnetic latex particles are added from the ancillary well. The Lite Reagent, packaged in a ReadyPack ancillary reagent pack, contains a biotinylated anti-HBs mouse monoclonal capture antibody and an acridinium-ester labeled anti-HBs mouse monoclonal antibody. HBsAg in the sample complexes with the antibodies and Streptavidin coated magnetic latex particles in the Solid Phase capture the HBsAg-antibody complexes. The sample is incubated simultaneously with Solid Phase, Lite Reagent, and Ancillary Reagent. Antibody-antigen complexes will form if hepatitis B surface antigen is present in the sample. Sample is first incubated with the Lite Reagent for 6 minutes at 37°C. During this incubation biotinylated anti-HBs monoclonal antibody and acridinium conjugated anti-HBs monoclonal antibody bind to HBsAg present in the sample. The Solid Phase and non-magnetic microparticles are added next, and the streptavidin-coated microparticles in the Solid Phase bind the biotin of the sample HBsAg / acridinium conjugated anti-HBs / biotinylated anti-HBs complex during an 18 minute incubation at 37°C. The microparticles are then held fast by a magnet and washed multiple times to remove unbound sample. The reaction mix is next reacted with acid and base to initiate a chemiluminescent reaction of the bound acridinium ester. The chemiluminescent signal is detected and quantified as relative light units (RLUs) by the photomultiplier tube (PMT) of the ADVIA Centaur® Instrument. The relative light units (RLUs) detected by the ADVIA Centaur® system are used to calculate the Index Value from the Master Curve. The RLU value is compared to a stored calibration curve to generate a patient result. The ADVIA Centaur® HBsAg Confirmatory assay uses the principle of specific antibody neutralization to confirm the presence of HBsAg in a sample that is repeatedly reactive in the HBsAg assay. The sample is pretreated and tested in parallel; one sample aliquot is dispensed and incubated with a neutralizing reagent containing high titers of anti-HBs (Reagent A); the second sample aliquot is incubated with a non-neutralizing control reagent (Reagent B). HBsAg in the patient sample is bound by the anti-HBs in Reagent A and not allowed to react in the ADVIA Centaur® HBsAg assay. When both aliquots are run in the ADVIA Centaur® HBsAg assay, the inhibition of the RLU signal in the aliquot with Reagent A is compared to the RLU signal in the aliquot with Reagent B. The relative percent neutralization is calculated and an interpretation of the sample is generated. If the RLU value with Reagent B is below the cut off, the assay is invalid. If percent neutralization is &gt;50% the sample is confirmed positive for HBsAg. 3 {3} Summary of Safety and Effectiveness ## Calibration The ADVIA Centaur® HBsAg assay utilizes a factory set Master Curve. The Master Curve values are contained on the Master Curve card provided with each kit. The master curve and calibration are lot specific. The barcode reader or keyboard is used to enter the Master Curve values on the system. The 2 calibrators in the kit are run when the lot is first used or after expiration of the calibration interval (21 days). If the calibration run is valid, as determined by prearranged parameters, the values are stored and used to “normalize” test values to the Master Curve. The ADVIA Centaur® HBsAg Confirmatory assay utilizes the same calibrators as the HBsAg assay but has its own Master Curve card. ## Assay antigen/antibody Description ### HBsAg Assay The ADVIA Centaur® HBsAg assay utilizes biotinylated anti-HBs mouse monoclonal antibody (directed to the HBsAg “a” region determinant) and AE-labeled anti-HBs mouse monoclonal antibody in the Lite Reagent. ### HBsAg Confirmatory Assay The basis for the ADVIA Centaur® HBsAg Confirmatory assay is specific antibody neutralization prior to antibody/antigen interaction. An aliquot of the sample to be tested is pretreated with high titers of antibody to HBsAg (human anti-HBs; Reagent A) and then tested in the ADVIA Centaur® HBsAg assay. In parallel, an aliquot of the sample is pretreated by incubation with non-neutralizing control reagent (decalcified human plasma) that does not contain high titers of antibody to HBsAg (Reagent B). Hepatitis B virus surface antigen in the patient sample is bound by the anti-HBs in Reagent A and is not allowed to react in the ADVIA Centaur® HBsAg assay. ## VI. ALTERNATIVE PRACTICES OR PROCEDURES Determination of the presence of HBsAg in patients may be achieved by using a number of commercially available, FDA licensed/approved, serological tests. When the results of such tests are evaluated in conjunction with a physician’s assessment and other biochemical test results, a diagnosis of infection with HBV can be established. ## VII. MARKETING HISTORY The ADVIA Centaur® HBsAg and ADVIA Centaur® HBsAg Confirmatory Assays are currently being marketed internationally in accordance with section 802 of the Food Drug and Cosmetic act in the following countries: Colombia, Sweden, Norway, Finland, France, Germany, Italy, Spain, Portugal, United Kingdom, Belgium, Austria, South Africa, Japan, China, Hong Kong, Singapore, Malaysia, Korea, Australia, and New Zealand. This product has not been withdrawn from any country for any reason. ## VIII. POTENTIAL ADVERSE EFFECTS OF THE DEVICE ON HEALTH The ADVIA Centaur® HBsAg ReadyPack Reagents, ADVIA Centaur® HBsAg Confirmatory Reagents, and ADVIA Centaur® HBsAg Calibrators are for in vitro diagnostic use, thus there is no direct adverse effect on the patient. 12 {4} Summary of Safety and Effectiveness Failure of the product to perform as intended, or errors in the use of the product, may lead to a false result. ## ADVIA Centaur® HBsAg Assay A false reactive (false positive) result with an HBsAg assay is not usually considered a patient or public health concern. A reactive result in the clinical lab setting is usually accompanied with results for additional HBV or biochemical markers or followed up with supplemental testing such as neutralization. In screening of pregnant women, there is a potential consequence to the newborn of receiving unnecessary therapy. If the mother tests positive for HBsAg, it is recommended that the newborn be given hepatitis B immune globulin (HBIG) and HBV vaccine within 12 hours of birth. A false positive result would therefore expose the newborn to the risk of receiving human source material. A false non-reactive (false negative) result with an HBsAg assay is a concern primarily in screening of pregnant women. A false negative test would mean that a newborn may not receive treatment and would be at higher risk of acute or chronic HBV infection. No HBsAg assay can guarantee to detect all infectious individuals. An individual may have antigen levels too low to be detected or the antigen produced may not be detectable due to genetic mutation. Because of the complexity of the mutations that can occur, no manufacturer can guarantee to detect all infectious donors and patients. ## ADVIA Centaur® HBsAg Confirmatory Assay A false “confirmed” result when using an HBsAg Confirmatory assay is not considered a patient or public health concern because the results in both the neutralized sample and the non-neutralized control would have to be incorrect to report a false confirmation. If this did occur the implications would be the same as for the HBsAg test. A false negative (not confirmed) result when using a HBsAg Confirmatory assay could occur due to a falsely increased signal in the neutralized sample, perhaps due to sample integrity or other malfunction, which would give the appearance that the sample had not neutralized. Were this to occur, the implication would be the same as for the HBsAg assay. ## IX. SUMMARY OF PRECLINICAL STUDIES ### Laboratory Studies The objectives of the laboratory studies were to test the analytical sensitivity, dynamic range, potential cross-reactive specimens, endogenous interferences, high dose hook effects, precision, matrix/collection and tube type effects, sample handling, stability, microbial studies, and instrument studies for performance characteristics of the ADVIA Centaur® HBsAg and ADVIA Centaur® HBsAg Confirmatory assays. ### Analytical Sensitivity The sensitivity of the ADVIA Centaur® HBsAg assay was evaluated using the WHO 1st International Reference Standard, low titer performance panels, and ad subtype and ay subtype HBsAg Sensitivity Panels. Dilutions of the WHO 1st International Reference Standard 80/549-1 were tested with multiple reagent lots. A concentration of 0.1 IU/mL was consistently positive with all lots while a concentration of 0.05 IU/mL was always negative. By regression analysis the HBsAg concentration at the cut off (Index = 1.0) was 0.066 IU/mL (95% CL = 0.0658 – 0.0662). Four commercially available low titer panels were used to assess the sensitivity of the assay for {5} Summary of Safety and Effectiveness known positive samples in the assay cut off range. The panels were tested with multiple reagent lots and the ADVIA Centaur® HBsAg assay was reactive on 100% of the positive panel members. The ad and ay subtype sensitivity panels were evaluated with multiple reagent lots. For the ad subtype sensitivity panel, HBsAg concentrations greater than or equal to 0.05 Paul Ehrlich Institute (PEI) Units/mL were detected 100% of the time. HBsAg concentrations of less than or equal to 0.03 PEI Units/mL were detected 0% of the time. The HBsAg concentration at the assay's assigned cut-off (1.0 Index) was estimated from the linear regression analysis as 0.034 PEI Units/mL (95% CL = 0.028 -0.040). For the ay subtype sensitivity panel, HBsAg concentrations greater than or equal to 0.03 Paul Ehrlich Institute Units/mL were detected 100% of the time. HBsAg concentrations of less than or equal to 0.02 PEI Units/mL were detected 0% of the time. The HBsAg concentration at the assay's cut-off (1.0 Index) was estimated from the linear regression analysis as 0.033 PEI Units/mL (95% CL 0.032 - 0.034). The following table summarizes these testing results: | Series | Cutoff (Index = 1.0) | 95% Exact Confidence Interval | | --- | --- | --- | | WHO 1^{st} IRP Standard, 80/549 | 0.066 IU/mL | 0.0658 – 0.0662 | | Commercially available HBsAg Sensitivity panel (ad subtype) | 0.034 PEI Units/mL | 0.028 – 0.04 | | Commercially available HBsAg Sensitivity panel (ay subtype) | 0.033 PEI Units/mL | 0.032 – 0.034 | The ADVIA Centaur® HBsAg assay was tested with an HBV genotype panel to demonstrate performance against HBV genotypes A-F. All 17 panel members were detected by the assay. To demonstrate confirmation of low positive samples by the ADVIA Centaur® HBsAg Confirmatory assay, 11 commercially available seroconversion panels and one low titer panel were tested. Of the 62 seroconversion panel samples that were reactive on the ADVIA Centaur® HBsAg assay, 59 were confirmed positive; the other 3 being “invalid” (control reagent was below the cut off, these specimens were not repeated). All the detected positive samples on the low titer panel were confirmed positive on ADVIA Centaur® HBsAg Confirmatory assay. The above information demonstrates the ability of the ADVIA Centaur® HBsAg to be analytically sensitive for the detection of HBsAg. ## Potential Cross-Reactive Specimens The specificity of the ADVIA Centaur® HBsAg assay was evaluated by testing 205 distinct serum specimens from 22 potentially cross-reacting groups. Patient samples from the following groups were tested: HAV antibody, HCV antibody, VZV, CMV, EBV, HSV-1, Syphilis, Toxoplasma IgM, Toxoplasma IgG, Rubella, Non-viral Liver Disease, Human anti-mouse antibodies (HAMA), Systemic Lupus Erythematosus, Rheumatoid factor, and Anti-nuclear antibody (ANA) specimens. Of these 22 sub-groups, 202 out of 205 specimens were observed to be non-reactive. One ANA specimen and 2 anti-HIV positive specimens were reactive. The following table shows testing result summary: 14 {6} Summary of Safety and Effectiveness | HBsAg Cross Reactor Summary Table | | | | | --- | --- | --- | --- | | Sample Group | Number Tested | Centaur® HBsAg assay non-reactive | Centaur® HBsAg assay reactive | | Rheumatoid Factor | 9 | 9 | 0 | | ANA – SLE | 7 | 6 | 1* | | Rubella IgG+ | 10 | 10 | 0 | | Non Viral Liver Disease | 10 | 10 | 0 | | HAV total | 5 | 5 | 0 | | HCV IgG+ | 10 | 10 | 0 | | VZV IgG+ | 10 | 10 | 0 | | CMV IgG+ | 10 | 10 | 0 | | HAMA | 9 | 9 | 0 | | Flu vaccine recipient | 10 | 10 | 0 | | Toxoplasma IgM | 7 | 7 | 0 | | Toxoplasma IgG | 8 | 8 | 0 | | EBV IgG+ | 10 | 10 | 0 | | EBV IgM+ | 10 | 10 | 0 | | Syphilis | 10 | 10 | 0 | | HSV-1 IgG+ | 10 | 10 | 0 | | HSV-1 IgM+ | 10 | 10 | 0 | | HCV + | 10 | 10 | 0 | | HIV + | 10 | 8 | 2** | | Rubella IgM+ | 10 | 10 | 0 | | CMV IgM+ | 10 | 10 | 0 | | VZV IgM+ | 10 | 10 | 0 | | Total Samples | 205 | 202 | 3 | * One sample was repeatedly reactive and not confirmed with the ADVIA Centaur® HBsAg Confirmatory assay ** Two Samples were repeatedly reactive and confirmed positive for HBsAg with the ADVIA Centaur® HBsAg Confirmatory assay In order to determine whether organisms other than HBV would cross-react or interfere with the ADVIA Centaur® HBsAg assay the following organisms, at the concentrations listed, were spiked into HBsAg nonreactive and HBsAg reactive specimens. | HBsAg results of various bacterial spikes | | | | --- | --- | --- | | Spike Material | Reactivity before spike | Reactivity after spike | | S. aureus 1,000 CFU/mL | Non-reactive | Non-reactive | | S. aureus 10,000 CFU/mL | Non-reactive | Non-reactive | | P. aeruginosa 1,000 CFU/mL | Non-reactive | Non-reactive | | P. aeruginosa 10,000 CFU/mL | Non-reactive | Non-reactive | | E. coli 1,000 CFU/mL | Non-reactive | Non-reactive | | E. coli 10,000 CFU/mL | Non-reactive | Non-reactive | | S. aureus 1,000 CFU/mL | Reactive | Reactive | | S. aureus 10,000 CFU/mL | Reactive | Reactive | | P. aeruginosa 1,000 CFU/mL | Reactive | Reactive | | P. aeruginosa 10,000 CFU/mL | Reactive | Reactive | | E. coli 1,000 CFU/mL | Reactive | Reactive | | E. coli 10,000 CFU/mL | Reactive | Reactive | IS {7} Summary of Safety and Effectiveness | HBsAg results of various viral antigen spikes | | | | --- | --- | --- | | Spike Material | Reactivity before spike | Reactivity after spike | | Epstein-Barr Virus 1 μg/mL | Non-reactive | Non-reactive | | Epstein-Barr Virus 1 ng/mL | Non-reactive | Non-reactive | | Cytomegalovirus 1 μg/mL | Non-reactive | Non-reactive | | Cytomegalovirus 1 ng/mL | Non-reactive | Non-reactive | | Rubella 1 μg/mL | Non-reactive | Non-reactive | | Rubella 1 ng/mL | Non-reactive | Non-reactive | | Varicella Zoster Virus 1 μg/mL | Non-reactive | Non-reactive | | Varicella Zoster Virus 1 ng/mL | Non-reactive | Non-reactive | | Epstein-Barr Virus 1 μg/mL | Reactive | Reactive | | Epstein-Barr Virus 1 ng/mL | Reactive | Reactive | | Cytomegalovirus 1 μg/mL | Reactive | Reactive | | Cytomegalovirus 1 ng/mL | Reactive | Reactive | | Rubella 1 μg/mL | Reactive | Reactive | | Rubella 1 ng/mL | Reactive | Reactive | | Varicella-Zoster Virus 1 μg/mL | Reactive | Reactive | | Varicella-Zoster Virus 1 ng/mL | Reactive | Reactive | The submitted information shows that the ADVIA Centaur® HBsAg assays should have minimal, if any, cross-reactivity with the specimens that contain the organisms at or below the concentrations tested. Several organisms that cause hepatitis disease were not tested, e.g., hepatitis A virus, parvovirus B19, and hepatitis E virus. Therefore, the user should be aware that it is unknown whether these organisms will cross-react or interfere with the ADVIA Centaur® HBsAg assays. It is believed that this issue is adequately covered by the labeling listing for the organisms tested. ## Endogenous Interferents The potentially interfering effects of conjugated bilirubin, unconjugated bilirubin, hemoglobin, triglycerides, hyper IgG, and low protein were evaluated using 10 specimens following the guidelines described by CLSI EP7-P¹ for interference due to endogenous substances. The effects of conjugated bilirubin at 40 mg/dL, unconjugated bilirubin at 40 mg/dL, hemoglobin at 500 mg/dL, triglycerides at 1000 mg/dL, hyper IgG at 6 g/dL (i.e., high total protein), and low protein at 3 g/dL were evaluated in serum specimens. No interference was found. ## High Index Hook Effect The Centaur® HBsAg assay was evaluated for decrease of Index value in specimens containing extremely high HBsAg levels by spiking purified HBsAg (ad and ay subtypes) into negative serum and preparation of a series of dilutions with each subtype. It was determined that the ADVIA Centaur® HBsAg reagents will not hook below the cut-off value (index&lt;1.0) at the HBsAg concentrations tested (approximately 5 mg/mL) for either the ad or ay subtypes. ## Precision Studies A single instrument CLSI EP5-A² precision study was done at Bayer Diagnostics using one lot of ADVIA Centaur® HBsAg Reagents. The study was set up as both an evaluation of HBsAg assay imprecision and a study of the effect of sample matrix (serum vs. five plasma anti- 16 {8} Summary of Safety and Effectiveness coagulants) on specimen precision. For each anti-coagulant, a single specimen was used to prepare a five member panel targeting the following HBsAg INDEX ranges (high HBsAg positive serum was used to prepare spiked positive panel members); 0.0 INDEX (negative, un-spiked) 0.5 INDEX (elevated negative, spiked) 2.0 INDEX (low positive #1, spiked) 4.0 INDEX (low positive #2, spiked) 20.0 INDEX (positive, spiked). Each sample was run twice a day in triplicate for 20 days. A new set of sample aliquots was thawed and mixed/centrifuged for each day of the study. Index values of the positive control, and spiked HBsAg specimen panels prepared in serum and three plasma types were calculated using a stored curve established by 2-point calibrator adjustment of the reagent master curve. The within-run and total assay precision (%CVs) of all samples with an Index &gt; 1.0 (cut-off value) ranged from 1.8% to 4.7% and from 2.5% to 5.6% respectively. The following table shows summary study information: | Sample | Mean Index | Within-run | | Among run | | Among Date | | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | | Positive Control | 4.76 | 0.09 | 1.9 | 0.09 | 1.8 | 0.09 | 2.0 | 0.16 | 3.3 | | Serum 1 | 0.56 | 0.04 | 7.2 | 0.01 | 2.5 | 0.03 | 5.5 | 0.05 | 9.4 | | Serum 2 | 1.82 | 0.06 | 3.2 | 0.05 | 2.5 | 0.04 | 2.1 | 0.08 | 4.6 | | Serum 3 | 3.57 | 0.07 | 2.0 | 0.04 | 1.2 | 0.06 | 1.7 | 0.10 | 2.9 | | Serum 4 | 17.07 | 0.37 | 2.2 | 0.34 | 2.0 | 0.39 | 2.3 | 0.64 | 3.7 | | K2 EDTA 1 | 0.49 | 0.05 | 9.8 | 0.04 | 7.4 | 0.02 | 3.7 | 0.06 | 12.8 | | K2 EDTA 2 | 1.86 | 0.07 | 3.6 | 0.00 | 0.0 | 0.05 | 2.6 | 0.08 | 4.5 | | K2 EDTA 3 | 3.93 | 0.11 | 2.7 | 0.03 | 0.9 | 0.09 | 2.2 | 0.14 | 3.6 | | K2 EDTA 4 | 19.13 | 0.37 | 2.0 | 0.36 | 1.9 | 0.26 | 1.4 | 0.58 | 3.1 | | Lithium heparin 1 | 0.36 | 0.04 | 9.7 | 0.02 | 6.6 | 0.03 | 7.0 | 0.05 | 13.7 | | Lithium heparin 2 | 1.77 | 0.08 | 4.6 | 0.04 | 2.1 | 0.04 | 2.5 | 0.10 | 5.6 | | Lithium heparin 3 | 3.77 | 0.08 | 2.1 | 0.06 | 1.6 | 0.06 | 1.6 | 0.11 | 3.0 | | Lithium heparin 4 | 16.92 | 0.30 | 1.8 | 0.20 | 1.2 | 0.24 | 1.4 | 0.43 | 2.5 | | Sodium heparin 1 | 0.42 | 0.50 | 11.2 | 0.02 | 3.8 | 0.02 | 5.4 | 0.05 | 13.0 | | Sodium heparin 2 | 2.08 | 0.10 | 4.7 | 0.04 | 1.8 | 0.00 | 0.0 | 0.10 | 5.0 | | Sodium heparin 3 | 3.72 | 0.10 | 2.8 | 0.07 | 1.9 | 0.00 | 0.0 | 0.13 | 3.4 | | Sodium heparin 4 | 19.87 | 0.39 | 2.0 | 0.43 | 2.2 | 0.20 | 1.0 | 0.62 | 3.1 | ## Matrix, Collection Tube Type, Effects Collection Tube Type Blood was collected from 50 individuals into serum glass, serum plastic, K₂EDTA plastic, lithium heparin glass, lithium heparin plastic, sodium heparin glass, sodium heparin plastic, serum separator tube (SST) glass, and SST plastic. All serum and plasma donors were initially screened to be HBsAg and anti-HBs negative. Specimens used for positive spikes were screened to be anti-HBs negative. Using a high titer HBsAg plasma pool, specimens were spiked to reach HBsAg target levels covering low cut-off positives (2 - 8 ADVIA Centaur® Index (Index)) and high positive samples (50 - 150 Index). The distribution of the samples included in the study was twenty negatives, eleven low cut-off positives, and nineteen high 9 17 {9} Summary of Safety and Effectiveness 10 18 positives. For evaluation purposes, the positive specimens were grouped into five groups with the following serum glass index ranges for each group: | | n | Serum Glass Min^{1} Index | Serum Glass Max^{2} Index | Serum Glass Mean Index | Serum Plastic Min Index | Serum Plastic Max Index | Serum Plastic Mean Index | LiHeparin^{3} Glass Min Index | LiHeparin Glass Max Index | LiHeparin Glass Mean Index | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Group 1 | 12 | 2.1 | 3.6 | 2.9 | 2.1 | 3.7 | 2.9 | 1.7 | 3.2 | 2.6 | | Group 2 | 10 | 7.5 | 8.1 | 7.8 | 7.2 | 8.1 | 7.8 | 6.6 | 7.5 | 7.2 | | Group 3 | 20 | 53.6 | 68.9 | 59.9 | 53.9 | 81.5 | 60.9 | 50.8 | 64.9 | 57.3 | | Group 4 | 10 | 103.0 | 115.6 | 108.8 | 109.6 | 123.0 | 116.0 | 100.0 | 134.2 | 114.6 | | Group 5 | 8 | 125.2 | 143.5 | 136.3 | 119.5 | 132.4 | 127.2 | 104.1 | 126.8 | 113.6 | 1 = Minimum 2 = Maximum 3 = Lithium Heparin Results for the serum glass specimens were considered the reference results. Bias for the various specimen collection devices were determined by calculating the index percent difference from serum glass for each individual result. These results were then graphed with the x-axis showing serum index result and the y-axis showing percent difference from serum. The following two graphs show the results for serum plastic and sodium lithium glass. ![img-0.jpeg](img-0.jpeg) {10} Summary of Safety and Effectiveness ![img-1.jpeg](img-1.jpeg) Based on the results obtained it was determined that the following blood collection tubes are acceptable for use in the ADVIA Centaur® HBsAg and HBsAg Confirmatory assays: serum plastic, K₂EDTA plastic, serum separator tube (SST) glass, and SST plastic. For these anti-coagulants, the specimen index did not appreciably vary as a function of collection tube type. Due to the variation noted with lithium heparin glass, lithium heparin plastic, sodium heparin glass, and sodium heparin plastic it was determined that specimens collected with these devices that produced results ≥ 0.7 to 0.99 required additional testing, which may include other HBV serological marker and supplemental tests. ## Sample Handling Studies The sample handling studies are a series of experiments in which specimens collected in all of the sample matrices claimed as suitable for use with the Centaur® HBsAg assay are subjected to potential stresses such as freeze/thaw or elevated temperature storage and tested in comparison to baseline data to determine the impact of the stress on assay accuracy. The sample handling studies described here evaluate the effect of the following patient sample handling conditions on ADVIA Centaur® HBsAg Index: 1. Extended time on-board the Centaur® Instrument (simulated by non-capped storage at 25°C). 2. Extended time in refrigerated (2-8°C) storage. 3. Extended time at room temperature (25°C) storage. 4. Extended time in freezer (-20°C) storage. 5. Multiple freeze/ thaw (-20°C/2-8°C) cycles. Samples were collected during in-house blood draws from healthy donors in serum and plasma with a variety of anti-coagulants. The serum and plasma samples were aliquoted and placed in appropriate storage/stress conditions on the day of collection. A baseline Index value for each sample was established by testing with the ADVIA Centaur® HBsAg assay on the day of collection. All percent recoveries were calculated against this baseline (day 0) value. Results from the ADVIA Centaur® HBsAg sample handling studies support the claims that samples can 19 {11} Summary of Safety and Effectiveness be subjected to the following conditions and still generate accurate results when tested in the ADVIA Centaur® HBsAg assay: 1. Samples can be kept on-board the Centaur® instrument for up to 8 hours. 2. Samples can be stored refrigerated (2-8°C) for up to 14 days. 3. Samples can be stored at room temperature for up to 8 hours. 4. Samples can be stored frozen (-20°C) for long term storage. 5. Samples can be frozen and thawed up to six times. ## On-The-Cells Specimen Storage A study was done to determine if storing the processed serum or plasma sample in the original collection tube (“On the Cells”) rather than transferring the serum or plasma to a secondary container affects the ADVIA Centaur® HBsAg Index. Fresh samples were drawn from 10 healthy in-house donors into serum and plasma collection tube types. The specimens were centrifuged within the specified study times and an aliquot from each primary tube placed in another container (HBsAg positive specimens were spiked with high positive HBsAg pool before removing the aliquot). The samples were then stored at 2-8°C and tested side by side (on-the-cells versus plasma or serum) in the ADVIA Centaur® HBsAg assay at 1, and 3 days after collection. There is no evidence of any effect on the final assay result for the following specimen types: serum plastic, K₂EDTA plastic, sodium heparin glass, sodium heparin plastic, serum separator tube (SST) glass, and SST plastic due to sample being left in the primary collection tube for up to three days of 2-8°C storage. The study show that specimens collected in lithium heparin (glass and plastic) may be left in the primary collection tube for up to 12 hours at 2-8°C. ## Sample Processing – Time to Centrifugation A study was done to determine the effect of fresh sample time to centrifugation on the HBsAg Index. Ten (10) specimens were drawn from healthy in-house donors in serum collection tubes. The tubes were spiked with HBsAg positive pool and centrifuged shortly after collection and after 24 hours. Aliquots from the tubes were then tested on the ADVIA Centaur® to determine if leaving blood unprocessed in the collection tube for as long as 24 hours caused the specimen’s ADVIA Centaur® HBsAg index to shift. There was no apparent change in clinical results, i.e., reactive or nonreactive, when centrifugation was delayed up to 24 hrs. ## Sample Handling – Inversion of Gel barrier Collection tubes A study was done to determine if inversion of barrier gel blood collection tubes interferes with ADVIA Centaur® HBsAg assay results. Blood was drawn from 22 healthy in-house donors using serum and plasma (Lithium Heparin) gel barrier (separator) collection tubes. Seven sample tubes were spiked with HBsAg at different levels, three were left negative and 12 specimens were spiked to levels close to the 1.0 Index cut-off. The tubes were rocked for 30 minutes, centrifuged, and an aliquot was taken. The tubes were then inverted 5 times and a second aliquot was taken. The two aliquots were compared to determine if inversion altered sample Index. Inversion of the barrier gel collection tube, 5 times, had no apparent effect on the assay results for both positive and negative samples. ## Stability Studies 20 {12} Summary of Safety and Effectiveness Three lots of HBsAg reagents (Centaur® ReadyPack reagents, calibrators, and controls) were used for real time stability studies. All kits and reagents were stored at the recommended storage temperature of 2-8°C. Reagents and calibrators were monitored at several check points post manufacturing date. The submitted shelf-life studies support a claim of a 52 week expiration dating at 2-8°C for the HBsAg reagents. Three lots of the ADVIA Centaur® HBsAg Confirmatory Reagents were placed on real time stability studies. All kits and reagents were stored at the recommended storage temperature of 2-8°C. Reagents were monitored at several checkpoints post manufacturing date. The submitted shelf-life studies support a claim of a 104 week expiration dating at 2-8°C for the HBsAg Confirmatory Reagents. To determine the shelf life for assay controls and calibrators, three lots of calibrators and three lots of controls have been placed on 2-8°C long-term stability. All calibrators and controls were stored at the recommended storage temperature of 2-8°C. Calibrators and controls were monitored at several checkpoints post manufacturing date. The submitted shelf-life studies support a claim of 12 month expiration dating at 2-8°C for the ADVIA Centaur® HBsAg calibrators and controls. Reagent On-Board Stability (OBS) and Calibration Interval Studies: Two lots of reagents have undergone reagent OBS studies on two Centaur® instruments. OBS testing on the instruments occurred at several checkpoints after the reagents are placed on-board. A fresh pack serves as the control for each time-point. It was determined that a dose recovery within 10% or 2SD of the fresh pack serves to define acceptable performance. A calibration interval of 21 days was also evaluated using these results. The submitted on-board studies for the reagents support 41 days OBS (999 hours) for the ADVIA Centaur® HBsAg reagents. The OBS studies also support a re-calibration interval of 21 days. To support reagent shipping recommendations shipping studies of the reagents found that the HBsAg reagents could withstand 3x freeze/thaw cycles (-40°C to 4C) without aggregation of the solid phase and acceptable assay performance. The recommended shipping conditions are to ship the ReadyPack reagents in an upright position and stored at 2-8°C. A calibrator and control shipping study was performed on one manufactured lot. The HBsAg Calibrators and controls underwent 3x freeze/thaw cycles with no adverse effects. The calibrator and control open vial study examined the length of time the calibrator or control is stable once the vial is opened. Open vials are stored at the recommended storage conditions of 2-8°C. The open vials are sampled periodically up to 90 days post initial opening. Fresh (unopened) vials are evaluated at each time point to serve as controls. The acceptance criterion for this study was dose recovery within 10% (or 2SD) of the fresh vial dose. The submitted study information supports an open vial use lifetime of up to 90 days. ## Microbiology Studies The ADVIA Centaur® HBsAg and HBsAg Confirmatory reagents contain amphotericin, gentamicin, and sodium azide as preservatives to protect against contamination by microorganisms. The ADVIA Centaur® calibrators and Negative Control contain 0.09% sodium azide and the Positive Control contains 0.25% Micro-O-Protect as preservatives to protect against contamination by microorganisms. The reagents and calibrators were challenged in a study conducted according to United States Pharmacopeia (USP) requirements for Antimicrobial Effectiveness testing to assess the ability of the reagents to withstand or control microbial contamination. Results indicated that the preservative systems for reagents and calibrators met the USP requirements for antimicrobial effectiveness testing. A performance microbial challenge was performed using one lot of ADVIA Centaur® HBsAg and HBsAg Confirmatory reagents, calibrators, and controls. Reagents and controls were 13 21 {13} Summary of Safety and Effectiveness inoculated with two pools of microbes containing the USP defined organisms at 10E3 and 10E6 CFU/mL and then run on the ADVIA Centaur® instrument at time 0 (Baseline), 30 days and 60 days. Controls, medical decision pools, and a quality control panel were all within expected ranges when tested using inoculated reagents at all timepoints. ## Instrument Studies Software and hardware verification testing were performed for the ADVIA Centaur. Appropriate information and study results were furnished demonstrating that the ADVIA Centaur® hardware and software, used with the ADVIA Centaur® HBsAg reagents, functioned as described and had appropriate safeguards ## Environmental Testing The purpose of environmental testing is to assess ADVIA Centaur® HBsAg and HBsAg Confirmatory assay control recovery or percent neutralization and interpretations at the mean and extreme environmental conditions as specified (18 to 30 °C). Each assay was calibrated and run on a single instrument unit in an environmental chamber. These studies demonstrated acceptable performance of the HBsAg and HBsAg Confirmatory assays when performed on instruments operating at the extremes of the temperature range for the ADVIA Centaur® instrument (18°C to 30°C). ## Reagent Compatibility Testing The purpose of this study was to confirm there are no primary reagent interactions for assays that share the same reagent probe, and might therefore be susceptible to reagent carryover affects. Mitigation of any interference identified is accomplished through Test definition scheduling options, using multiple water washes, or, in rare occasions where a Wash Pack with a solution other than water may be required. The ADVIA Centaur® HBsAg assay and HBsAg Confirmatory assay were evaluated for their potential affect on all other assays using the same reagent probes and for the affect of all the other assay reagents on the HBsAg or HBsAg Confirmatory assay. To be accepted there must be &lt; 5% difference in Index (or dose) between test and control, or no statistically significant change in Index (or dose), or no more than 1SD difference in Index (or dose) as appropriate for the assay and the control being tested. The submitted studies show that the HBsAg primary (ReadyPack) reagent has no compatibility issues with assays that share a common probe. It was demonstrated that the HBsAg ancillary reagent would interfere with some ADVIA Centaur® immunoassays. To correct this issue Ancillary Probe washes were instituted in the assay software to mitigate this interference. It was demonstrated that HBsAg Confirmatory Reagent A and Reagent B ancillary reagents have no compatibility issues with other assays. ## Conclusions Drawn from the Pre-Clinical Studies The ADVIA Centaur® HBsAg assay was evaluated to demonstrate performance claims for cross-reactivity, interference, precision, matrix type, specimen handling, and reagent stability. The results of the pre-clinical studies demonstrated that the ADVIA Centaur® HBsAg reagents and instrument were capable of detecting HBsAg and with the assay there was minimal adverse effect on performance when other factors were included. ## X. SUMMARY OF CLINICAL STUDIES The objective of this clinical study was to assess the efficacy of ADVIA Centaur® HBsAg for detecting hepatitis B surface antigen in human serum as presumptive evidence of an HBV infection. 14 22 {14} Summary of Safety and Effectiveness ## Study Design The safety and effectiveness of the ADVIA Centaur®® HBsAg and ADVIA Centaur®® HBsAg Confirmatory assays was determined by a laboratory study consisting of the following: - A study of prospectively obtained samples from patients who are either at risk for hepatitis B virus (HBV) infection (at least one risk factor indicated on the patient’s case report form (CRF)), exhibiting signs and/or symptoms of HBV infection (at least two signs/symptoms indicated on the patient’s CRF), or undergoing dialysis. These samples were tested using FDA approved or licensed HBV serological assays and their HBV status classified on the basis of the HBV serological marker results (HBV surface antigen (HBsAg), HBV e antigen (HBeAg), anti-HBV core antigen IgM (anti-HBc IgM) total anti-HBV core antigen (total anti-HBc), anti-HBe antigen (anti-HBe), and total anti-HBsAg (anti-HBs) – this testing was considered the reference testing) and the ADVIA Centaur®® HBsAg assay at the clinical trial sites. - A study of retrospectively collected samples, obtained from a commercial vendor, from HBV acutely infected and HBV chronically infected patients. Vendor assignment of HBV acute samples was based upon positive HBsAg and anti-HBc IgM assay results. Vendor assignment of HBV chronic samples was based upon the individual having positive HBsAg results for at least six months. Vendor assignment was verified by HBV marker testing at the clinical sites. These samples were tested with both the ADVIA Centaur®® HBsAg assay and a reference HBsAg assay at the clinical trial sites. - A study of prospectively obtained prenatal samples to determine the use of the ADVIA Centaur® HBsAg assay for prenatal testing. This population included pregnant women in the 1st, 2nd, and 3rd trimesters. These samples were tested with both the ADVIA Centaur®® HBsAg assay and a reference HBsAg assay at the clinical trial sites. - A study of retrospectively collected HBV seroconversion panels obtained from a commercial vendor. These panels were tested with both the ADVIA Centaur®® HBsAg assay and a reference HBsAg assay at the clinical trial sites. - A precision and reproducibility study in which a specimen panel was assayed over several days, at multiple clinical trial sites, and using multiple ADVIA Centaur®® HBsAg assay reagent lots. The results were analyzed to derive precision estimates. These studies are described in more detail below. ## Gender Bias The total ratio of men versus women in the high risk, signs and symptoms, and dialysis prospective study was 39.9% male and 60.1% female. It is considered that this is reflective of the underlying distribution of the disease for each given age group, ethnic group, stage of disease, and the inclusion of a specific prenatal study. If the prenatal study population is removed the distribution for males is 54% and females 46%, which is a more equal gender distribution. There appeared to be no selection bias on the basis of gender identified during the review process other than those specimens which were retrospectively selected and the prenatal prospective population. There appeared to be no difference in the safety and effectiveness of the device based on gender. This device appeared to have a similar effectiveness between men and women. 15 23 {15} Summary of Safety and Effectiveness ## Demographic Data 2740 patient samples were with the ADVIA Centaur® HBsAg assay. The following results classified by gender and age range were obtained: | Bayer ADVIA Centaur® HBsAg Assay Distribution of High Risk, Signs and Symptoms, Dialysis, and Prenatal Population by Age Group and Gender All Testing Sites | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | | Age (years) | Gender | Positive^{a} | | Nonreactive^{b} | | Total | | | | | N | % | N | % | N | % | | 0-9 | Male | 0 | -- | 0 | -- | 0 | -- | | | Female | 0 | -- | 0 | -- | 0 | -- | | | Overall | 0 | -- | 0 | -- | 0 | -- | | 10-19 | Male | 1 | 14.3 | 6 | 85.7 | 7 | 6.6 | | | Female | 0 | -- | 99 | 100 | 99 | 93.4 | | | Overall | 1 | 0.9 | 105 | 99.1 | 106 | 100 | | 20-29 | Male | 6 | 7.1 | 78 | 92.9 | 84 | 14.7 | | | Female | 7 | 1.4 | 479 | 98.6 | 486 | 85.3 | | | Overall | 13 | 2.3 | 557 | 97.7 | 570 | 100 | | 30-39 | Male | 18 | 9.2 | 178 | 90.8 | 196 | 32.7 | | | Female | 16 | 4.0 | 388 | 96.0 | 404 | 67.3 | | | Overall | 34 | 5.7 | 566 | 94.3 | 600 | 100 | | 40-49 | Male | 35 | 9.1 | 350 | 90.9 | 385 | 54.5 | | | Female | 8 | 2.5 | 314 | 97.5 | 322 | 45.5 | | | Overall | 43 | 6.1 | 664 | 93.9 | 707 | 100 | | 50-59 | Male | 29 | 10.1 | 259 | 89.9 | 288 | 58.2 | | | Female | 5 | 2.4 | 202 | 97.6 | 207 | 41.8 | | | Overall | 34 | 6.9 | 461 | 93.1 | 495 | 100 | | 60-69 | Male | 9 | 10.3 | 78 | 89.7 | 87 | 46.5 | | | Female | 1 | 1.0 | 99 | 99.0 | 100 | 53.5 | | | Overall | 10 | 5.4 | 177 | 94.7 | 187 | 100 | | ≥70 | Male | 4 | 8.9 | 41 | 91.1 | 45 | 61.6 | | | Female | 0 | -- | 28 | 100 | 28 | 38.4 | | | Overall | 4 | 5.5 | 69 | 94.5 | 73 | 100 | | Unknown | Male | 0 | -- | 1 | 100 | 1 | 50.0 | | | Female | 0 | -- | 1 | 100 | 1 | 50.0 | | | Overall | 0 | -- | 2 | 100 | 2 | 100 | | Total | Male | 102 | 9.3 | 991 | 90.7 | 1093 | 39.9 | | | Female | 37 | 2.2 | 1610 | 97.8 | 1647 | 60.1 | | | Overall | 139 | 5.1 | 2601 | 94.9 | 2740 | 100 | a Samples with an Index Value ≥ 1.00 that were confirmed positive b Samples with an Index Value &lt; 1.00 and initially reactive samples that did not confirm as positive ## Data Analysis and Results ## Prospective Study The prospective study population for the ADVIA Centaur® HBsAg assay and the ADVIA Centaur® HBsAg confirmatory assay consisted of 2740 patients. Of these 2740 patients, 965 patients (35.2%) were from a population considered at risk for hepatitis (high risk) due to lifestyle, behavior, occupation, or known exposure events, 847 patients (30.9%) were from the population of individuals exhibiting signs and/or symptoms of hepatitis infection (signs and 24 {16} Summary of Safety and Effectiveness symptoms), 212 patients (7.7%) were from the dialysis population, and 716 patients (26.1%) were from the prenatal population. The prenatal population included serum samples from healthy, pregnant women in their first trimester (149 of 716 patients [20.8%]), second trimester (272 of 716 patients [38.0%]), or third trimester (295 of 716 patients [41.2%]) of pregnancy. The prospective study population was 38.4% Hispanic, 31.8% Caucasian, 22.6% Black, 2.9% Asian, and 4.3% from unknown or other ethnicity. The majority of patients were female (60.1% female and 39.9% male). The mean age was 41.0 years (range of 12 to 82 years). Patients in the prospective study population were from the following geographic regions: Florida (37.8%), Texas (33.8%), New York (22.2%), and California (6.2%). The HBV disease classification for each patient in the high risk, signs and symptoms, and dialysis populations (2024 total) was determined by serological assessment using resultant hepatitis marker profiles obtained from results of commercially available, FDA-approved assays, considered the reference assays. Testing of the specimens occurred at each study site. The individual ADVIA Centaur® HBV assay result was compared to the reference HBsAg assay result and to the patient classification determined by the six HBV serological marker combinations. No patients were excluded from the complete study set due to incomplete reference HBV serological results. Disease classification for each individual was based only on the HBV serological marker results, and was not affected by additional laboratory or clinical information. Results for HBsAg were based on the results of HBsAg neutralization testing for both the ADVIA Centaur® HBsAg assay and the reference assay. The unique reference marker patterns observed using the ADVIA Centaur® HBsAg assay are presented in the following table. 17 25 {17} Summary of Safety and Effectiveness | Bayer ADVIA Centaur® HBsAg Assay Classification by HBV Reference Markers (All Testing Sites) | | | | | | | | --- | --- | --- | --- | --- | --- | --- | | HBV Classification | HBV Reference Markers | | | | | | | | HBsAg | HBeAg | IgM Anti-HBc | Total Anti-HBc | Anti-HBe | Anti-HBs (>10 mIU/mL) | | Acute | + | + | + | + | + | - | | Acute | + | + | + | + | - | - | | Acute | + | - | + | + | + | - | | Chronic | + | + | - | + | + | - | | Chronic | + | + | - | + | - | + | | Chronic | + | + | - | + | - | - | | Chronic | + | - | - | + | + | + | | Chronic | + | - | - | + | + | - | | Chronic | + | - | - | + | - | + | | Chronic | + | - | - | + | - | - | | Chronic | + | + | + | + | - | + | | Early Recovery | - | - | + | + | + | + | | Early Recovery | - | - | + | + | + | - | | Early Recovery | - | - | + | + | - | + | | Early Recovery | - | - | - | + | + | - | | Recovery | - | - | - | + | + | + | | Recovery | - | - | - | - | + | + | | Recovered | - | - | - | + | - | + | | Recovered | - | - | - | + | - | - | | HBV Vaccine Response | - | - | - | - | - | + | | Not previously infected | - | - | - | - | - | - | | Uninterpretable | + | - | - | - | - | + | | Uninterpretable | + | - | - | - | - | - | | Uninterpretable | - | + | - | - | - | + | | Uninterpretable | - | + | - | - | - | - | | Uninterpretable | - | - | + | - | - | - | | Uninterpretable | - | + | - | + | - | + | | Uninterpretable | - | + | - | + | - | - | | Uninterpretable | - | + | - | + | + | + | + = Reactive - = Nonreactive Following the assignment of specimen classification, the HBsAg results (nonreactive and confirmed reactive) obtained using the ADVIA Centaur® method were compared with the nonreactive and confirmed reactive HBsAg results obtained using the reference method for each result category. The method comparison for patients in the high risk, signs and symptoms, and dialysis populations for all testing sites combined is presented in the following table. 26 {18} Summary of Safety and Effectiveness | Bayer ADVIA Centaur® HBsAg Assay Method Comparison in High Risk, Signs and Symptoms, and Dialysis Population by HBV Classification ADVIA Centaur® HBsAg Assay vs. HBsAg Reference Assay All Testing Sites | | | | | | | --- | --- | --- | --- | --- | --- | | HBV Classification | Reference HBsAg Negative | | Reference HBsAg Positive | | Total^{a} | | | ADVIA Centaur® HBsAg Assay | | ADVIA Centaur® HBsAg Assay | | | | | Positive | Nonreactive | Positive | Nonreactive | | | | N | N | N | N | N | | Acute | 0 | 0 | 10 | 1 | 11 | | Chronic | 0 | 0 | 109 | 2 | 111 | | Early Recovery | 0 | 126 | 0 | 0 | 126 | | Recovery | 1 | 210 | 0 | 0 | 211 | | Recovered | 3 | 320 | 0 | 0 | 323 | | HBV Vaccine Response | 0 | 384 | 0 | 0 | 384 | | Not Previously Infected | 2 | 826 | 0 | 0 | 828 | | Uninterpretable | 1 | 22 | 3 | 4 | 30 | | Total | 7 | 1888 | 122 | 7 | 2024 | a Total number of test results by HBV categories The percent agreement between the ADVIA Centaur® HBV method and the reference assays for each specimen classification was performed, including the upper and lower 95% confidence intervals. The positive, negative, and overall percent agreements were calculated as follows: Positive percent agreement: Number of ADVIA Centaur® HBsAg confirmed positive results in agreement with reference HBsAg X 100 Total number of reference HBsAg confirmed positive results Negative percent agreement: Number of ADVIA Centaur® HBsAg nonreactive results in agreement with reference HBsAg X 100 Total number of reference HBsAg nonreactive results Overall percent agreement: Number of ADVIA Centaur® HBsAg results in agreement with reference HBsAg X 100 Total number of reference HBsAg confirmed positive and nonreactive results The percent agreement between the ADVIA Centaur® HBsAg assay and the reference HBsAg assay for the high risk, signs and symptoms, and dialysis populations across all testing sites is summarized in the following table. 19 27 {19} Summary of Safety and Effectiveness | Bayer ADVIA Centaur® HBsAg Assay Percent Agreement and Confidence Intervals by HBV Classification in High Risk, Signs and Symptoms, and Dialysis Population ADVIA Centaur® HBsAg Assay vs. HBsAg Reference Assay All Testing Sites | | | | | | --- | --- | --- | --- | --- | | HBV Classification | Positive Percent Agreement % (x/n)a | 95% Exact Confidence Interval | Negative Percent Agreement % (x/n)b | 95% Exact Confidence Interval | | Acute | 90.91 (10/11) | 58.72 to 99.77 | -- | -- | | Chronic | 98.20(109/111) | 93.64 to 99.78 | -- | -- | | Early Recovery | -- | -- | 100.00 (126/126) | 97.11 to 100.00 | | Recovery | -- | -- | 99.53 (210/211) | 97.39 to 99.99 | | Recovered | -- | -- | 99.07 (320/323) | 97.31 to 99.81 | | HBV Vaccine Response | -- | -- | 100.00 (384/384) | 99.04 to 100.00 | | Not Previously Infected | -- | -- | 99.76 (826/828) | 99.13 to 99.97 | | Uninterpretable | 42.86 (3/7) | 9.90 to 81.59 | 95.65 (22/23) | 78.05 to 99.89 | | Overall | 94.57 (122/129) | 89.14 to 97.79 | 99.63 (1888/1895) | 99.24 to 99.85 | a x = the number of ADVIA Centaur® HBsAg results that are confirmed positive in agreement with the reference HBsAg confirmed positive results; n = the total number of reference HBsAg results that are confirmed positive b x = the number of ADVIA Centaur® HBsAg results that are nonreactive in agreement with the reference HBsAg; n = the total number of reference HBsAg results that are nonreactive ## Retrospective HBV Infected (acute and chronic stages of infection) Study A retrospective study was conducted using commercially obtained specimens, 49 well-characterized samples from patients diagnosed with acute HBV and 104 well-characterized samples from patients with chronic HBV (patients who had a positive HBsAg result at least 6 months prior to sample collection). Samples were evaluated using the ADVIA Centaur® HBV method and the reference assay). Vendor assignment of disease status was verified by testing with HBsAg, Anti-HBc IgM, Anti-HBc Total, and Anti-HBs assays at the testing sites. The method comparison for the acute and chronic retrospective population across all testing sites is presented in the following table. | Bayer ADVIA Centaur® HBsAg Assay Method Comparison in Retrospective HBV Infected Population ADVIA Centaur® HBsAg Assay vs. HBsAg Reference Assay All Testing Sites | | | | | | | --- | --- | --- | --- | --- | --- | | HBV Disease Classification | Reference HBsAg Negative | | Reference HBsAg Positive | | Total | | | ADVIA Centaur® HBsAg Assay | | ADVIA Centaur® HBsAg Assay | | | | | Positive | Nonreactive | Positive | Nonreactive | | | | N | N | N | N | N | | Acute | 0 | 0 | 49 | 0 | 49 | | Chronic | 0 | 0 | 102 | 2 | 104 | | Total | 0 | 0 | 151 | 2 | 153 | 20 {20} Summary of Safety and Effectiveness The percent agreement between the ADVIA Centaur® HBsAg assay and the reference HBsAg assay for the retrospective acute and chronic HBV infected population is summarized by testing site in the following table. | Bayer ADVIA Centaur® HBsAg Assay Percent Agreement and Confidence Intervals by Testing Sites in Retrospective HBV Population Bayer ADVIA Centaur® HBsAg Assay vs. HBsAg Reference Assay | | | | | | | --- | --- | --- | --- | --- | --- | | Testing Site | Population | Positive Percent Agreement % (x/n)^{a} | 95% Exact Confidence Interval | Negative Percent Agreement % (x/n)^{b} | 95% Exact Confidence Interval | | Testing Site 1 | Acute | 100 (14/14) | 76.8 to 100 | -- | -- | | | Chronic | 94.3 (33/35) | 80.8 to 99.3 | -- | -- | | Testing Site 2 | Acute | 100 (35/35) | 90.0 to 100 | -- | -- | | | Chronic | 100 (35/35) | 90.0 to 100 | -- | -- | | Testing Site 3 | Acute | -- | -- | -- | -- | | | Chronic | 100 (34/34) | 89.7 to 100 | -- | -- | | All Testing Sites | Acute | 100 (49/49) | 92.8 to 100 | -- | -- | | | Chronic | 98.1 (102/104) | 93.2 to 99.8 | -- | -- | a x = the number of ADVIA Centaur® HBsAg results that are positive in agreement with the reference HBsAg; n = the total number of reference HBsAg results that are positive b x = the number of ADVIA Centaur® HBsAg results that are nonreactive in agreement with the reference HBsAg; n = the total number of reference HBsAg results that are nonreactive The above studies demonstrate that if used according to the procedural directions the ADVIA Centaur® HBsAg and the ADVIA Centaur® HBsAg Confirmatory assay should provide a meaningful result for the diagnosis of HBV infection. ## Prospective Study in Pregnant Women Serum samples were prospectively collected from 716 healthy, pregnant women who were in the first, second, or third trimester of pregnancy. By testing these samples from pregnant women using the ADVIA Centaur® HBsAg Assay and the reference HBsAg assay, the performance of the ADVIA Centaur® HBsAg assay in identifying neonates who were at risk for HBV infection during the perinatal period was evaluated. Results of HBsAg testing (repeatedly nonreactive and confirmed positive by neutralization testing) were compared between the ADVIA Centaur® HBsAg assay and the reference HBsAg assay for the prenatal population. This comparison for all testing sites combined is presented in the following table. 21 {21} Summary of Safety and Effectiveness | Bayer ADVIA Centaur® HBsAg Method Comparison in Prenatal Population ADVIA Centaur® HBsAg Assay vs. HBsAg Reference Assay | | | | | | | --- | --- | --- | --- | --- | --- | | Population | Reference HBsAg Assay Negative | | Reference HBsAg Assay Positive | | | | | ADVIA Centaur® HBsAg Assay | | ADVIA Centaur® HBsAg Assay | | | | | Positive | Nonreactive | Positive | Nonreactive | Total | | | N | N | N | N | N | | 1st Trimester | 0 | 148 | 1 | 0 | 149 | | 2nd Trimester | 0 | 268 | 4 | 0 | 272 | | 3rd Trimester | 0 | 290 | 5 | 0 | 295 | | All | 0 | 706 | 10 | 0 | 716 | The percent agreement between the ADVIA Centaur® HBsAg assay and the reference HBsAg assay for the prenatal population by pregnancy trimester is summarized for all sites in the following table. | Bayer ADVIA Centaur® HBsAg Percent Agreement and Confidence Intervals by Testing Sites in Prenatal Population ADVIA Centaur® HBsAg Assay vs. HBsAg Reference Assay | | | | | | --- | --- | --- | --- | --- | | Trimester | Positive Percent Agreement % (x/n)^{a} | 95% Exact Confidence Interval | Negative Percent Agreement % (x/n)^{b} | 95% Exact Confidence Interval | | 1st | 100 (1/1) | 2.5 to 100 | 100 (148/148) | 97.5 to 100 | | 2nd | 100 (4/4) | 39.8 to 100 | 100 (268/268) | 98.6 to 100 | | 3rd | 100 (5/5) | 47.8 to 100 | 100 (290/290) | 98.7 to 100 | | All | 100 (10/10) | 69.2 to 100 | 100 (706/706) | 99.5 to 100 | a x = the number of ADVIA Centaur® HBsAg results that are confirmed positive in agreement with the reference HBsAg confirmed positive results; n = the total number of reference HBsAg results that are confirmed positive b x = the number of ADVIA Centaur® HBsAg results that are nonreactive in agreement with the reference HBsAg; n = the total number of reference HBsAg results that are nonreactive The above study supports the use of the ADVIA Centaur® HBsAg Assay for prenatal screening. ## Seroconversion Study To demonstrate when the ADVIA Centaur® HBsAg assay would first detect HBsAg after initial infection six commercially obtained seroconversion panels were tested with the ADVIA Centaur® HBsAg assay and a reference HBsAg assay. The table below lists the individual panel results. 30 {22} Summary of Safety and Effectiveness | | HBsAg Positive Result From Initial Draw Date | | Reference Assay vs ADVIA Centaur® Assay | | --- | --- | --- | --- | | Panel ID | Reference Assay (Days) | ADVIA Centaur® Assay (Days) | Difference in Bleed Numbers* | | 1 | 51 | 51 | 0 | | 2 | 42 | 42 | 0 | | 3 | 19 | 19 | 0 | | 4 | 12 | 9 | +2 | | 5 | 7 | 7 | 0 | | 6 | 61 | 61 | 0 | * The difference in bleed numbers is relative to the reference assay. For example, a +2 means that the reference assay required 2 additional bleeds before reactivity was determined as compared to the time-point when ADVIA Centaur® assay confirmed positive. Compared to the reference HBsAg assay, the ADVIA Centaur® HBsAg assay demonstrated efficacy for the detection of the appearance of HBsAg following HBV infection. To demonstrate the ability of the ADVIA Centaur® HBsAg Assay to detect truly positive HBsAg samples at ≥ 50 index value the ranges of index values for repeatedly reactive samples in the ADVIA Centaur® HBsAg assay were compared to neutralization results from the ADVIA Centaur® HBsAg Confirmatory assay. These data are summarized in the following table. | ADVIA Centaur® HBsAg Assay | Repeat Reactive Results | ADVIA Centaur® HBsAg Confirmatory Assay Confirmed | | | --- | --- | --- | --- | | Index Range | N | N (%) | 95% Exact Confidence Intervals | | ≤1 to <50 | 14 | 10 (71.40)* | 41.9 to 91.6 | | ≤50 to <500 | 27 | 27 (100) | 87.2 to 100 | | >500 | 102 | 102 (100) | 96.4 to 100 | | Total | 143 | 139 (97.20) | 93.0 to 99.2 | * Neutralization testing not performed on 4 specimens. The above data support the use of the ≥ 50 index cutoff for not performing neutralization testing for samples whose results fall within this range. The low specimen number within the range of 50 - &lt;300 is justified based on the low number of specimens that would fall within this range. It is believed that if this specimen number were larger the ADVIA Centaur® HBsAg assay would have a lower 95% Exact confidence interval value of ≥90%. There were 133 samples that were repeatedly reactive or had results ≥ 50 index using the ADVIA Centaur® HBsAg assay. These samples were tested for HBsAg neutralization using the ADVIA Centaur® HBsAg Confirmatory assay. The results of this testing, for all testing sites, by HBV classification are presented in the following table. 31 {23} Summary of Safety and Effectiveness | Confirmatory Results for Reactive Samples by HBV Classification in High Risk, Signs and Symptoms, and Dialysis Populations (All Testing Sites) | | | | | | --- | --- | --- | --- | --- | | HBV Classification | ADVIA Centaur® HBsAg Assay | | Reference EIA HBsAg Assay | | | | Repeat Reactive (n) | Positive By Neutralization (n)(%) | Reference assay Percent agreement with ADVIA Centaur® (%)^{1} (n/n)^{2} | | | Acute | 10 | 10 (100) | 100% (10/10) | 69.2 to 100 | | Chronic | 109 | 109 (100) | 100% (109/109) | 96.7 to 100 | | Early Recovery | 0 | 0 (0/0) | NA | NA | | Recovery | 1 | 1 (100) | 0% (0/1) | NA | | Recovered | 5 | 3 (60) | 0% (0/3) | 0.0 to 70.8 | | HBV Vaccine Response | 0 | 0 (0/0) | NA | NA | | Not Previously Infected | 4 | 2 (50) | 0% (0/2) | 0.0 to 84.2 | | Uninterpretable | 4 | 4(100) | 75.0% (3/4) | 19.4 to 99.4 | | Total | 133 | 129 (96.99) | 94.57% (122/129) | 89.1 to 97.8 | 1 The percentage of Reference EIA HBsAg positive by neutralization samples which were also positive by neutralization in the ADVIA Centaur® HBsAg assay. 2 The number of samples which were positive by neutralization in the Reference EIA HBsAg assay over the number of samples which were positive by neutralization in the ADVIA Centaur® HBsAg assay. ## Precision and Reproducibility Study The ADVIA Centaur® HBsAg assay precision and reproducibility study was performed at 3 external sites utilizing 2 reagent lots per site. A 5-member panel and controls were assayed in replicates of 5 on a single run per day over 6 days for each lot. The study was completed with a single calibration of the assay (one calibration interval). Standard deviation and percent CV were calculated for within run, between run, and total. The site-specific results are presented in the following table. 32 {24} Summary of Safety and Effectiveness | Bayer ADVIA Centaur®® HBsAg Assay Precision Estimates for All Testing Sites and Reagent Lots | | | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Panel Member | Mean ADVIA Centaur® HBsAg Index Value | Within Run^{a} | | Between Run^{b} | | Total^{c} | | No. of Observations | No. of Days | | | | SD | CV (%) | SD | CV (%) | SD | CV (%) | | | | Testing Site 1, Reagent Lot 85 | | | | | | | | | | | 1 | 0.090 | 0.000 | NA | 0.000 | NA | 0.000 | NA | 30 | 6 | | 2 | 0.625 | 0.086 | 13.76 | 0.173 | 27.69 | 0.193 | 30.92 | 30 | 6 | | 3 | 1.738 | 0.084 | 4.82 | 0.179 | 10.30 | 0.198 | 11.37 | 30 | 6 | | 4 | 4.315 | 0.122 | 2.83 | 0.185 | 4.28 | 0.221 | 5.13 | 30 | 6 | | 5 | 20.156 | 0.395 | 1.96 | 0.820 | 4.07 | 0.910 | 4.52 | 30 | 6 | | Negative Control | 0.090 | 0.000 | NA | 0.000 | NA | 0.000 | NA | 30 | 6 | | Positive Control | 4.460 | 0.112 | 2.51 | 0.351 | 7.86 | 0.368 | 8.25 | 30 | 6 | | Testing Site 1, Reagent Lot 86 | | | | | | | | | | | 1 | 0.090 | 0.000 | NA | 0.000 | NA | 0.000 | NA | 30 | 6 | | 2 | 0.558 | 0.058 | 10.32 | 0.128 | 22.99 | 0.141 | 25.20 | 30 | 6 | | 3 | 1.556 | 0.070 | 4.49 | 0.244 | 15.71 | 0.254 | 16.34 | 30 | 6 | | 4 | 3.932 | 0.088 | 2.24 | 0.337 | 8.57 | 0.348 | 8.86 | 30 | 6 | | 5 | 19.163 | 0.349 | 1.82 | 0.926 | 4.83 | 0.990 | 5.16 | 30 | 6 | | Negative Control | 0.090 | 0.000 | NA | 0.000 | NA | 0.000 | NA | 30 | 6 | | Positive Control | 4.337 | 0.097 | 2.23 | 0.293 | 6.76 | 0.308 | 7.11 | 30 | 6 | | Testing Site 2, Reagent Lot 87 | | | | | | | | | | | 1 | 0.090 | 0.000 | NA | 0.000 | NA | 0.000 | NA | 30 | 6 | | 2 | 0.671 | 0.083 | 12.42 | 0.150 | 22.42 | 0.172 | 25.63 | 30 | 6 | | 3 | 1.696 | 0.065 | 3.81 | 0.079 | 4.64 | 0.102 | 6.00 | 30 | 6 | | 4 | 4.270 | 0.106 | 2.49 | 0.099 | 2.33 | 0.145 | 3.41 | 30 | 6 | | 5 | 20.049 | 0.244 | 1.22 | 0.448 | 2.23 | 0.510 | 2.54 | 30 | 6 | | Negative Control | 0.093 | 0.009 | NA | 0.014 | NA | 0.017 | NA | 30 | 6 | | Positive Control | 4.742 | 0.110 | 2.32 | 0.261 | 5.50 | 0.283 | 5.97 | 30 | 6 | | Testing Site 2, Reagent Lot 86 | | | | | | | | | | | 1 | 0.094 | 0.021 | NA | 0.018 | NA | 0.027 | NA | 29 | 6 | | 2 | 0.764 | 0.062 | 8.15 | 0.106 | 13.86 | 0.123 | 16.08 | 30 | 6 | | 3 | 1.758 | 0.070 | 4.00 | 0.145 | 8.25 | 0.161 | 9.17 | 30 | 6 | | 4 | 4.127 | 0.129 | 3.13 | 0.281 | 6.80 | 0.309 | 7.48 | 30 | 6 | | 5 | 18.889 | 0.276 | 1.46 | 0.821 | 4.35 | 0.866 | 4.59 | 30 | 6 | | Negative Control | 0.189 | 0.061 | NA | 0.092 | NA | 0.111 | NA | 30 | 6 | | Positive Control | 4.565 | 0.117 | 2.56 | 0.334 | 7.33 | 0.354 | 7.76 | 30 | 6 | 33 {25} Summary of Safety and Effectiveness | Bayer ADVIA Centaur®® HBsAg Assay Precision Estimates for All Testing Sites and Reagent Lots (continued) | | | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Panel Member | Mean ADVIA Centaur® HBsAg Index Value | Within Run^{a} | | Between Run^{b} | | Total^{c} | | No. of Observations | No. of Days | | | | SD | CV (%) | SD | CV (%) | SD | CV (%) | | | | Testing Site 3, Reagent Lot 87 | | | | | | | | | | | 1 | 0.090 | 0.000 | NA | 0.000 | NA | 0.000 | NA | 30 | 6 | | 2 | 0.656 | 0.142 | 21.64 | 0.154 | 23.55 | 0.210 | 31.98 | 30 | 6 | | 3 | 1.689 | 0.132 | 7.82 | 0.227 | 13.47 | 0.263 | 15.58 | 30 | 6 | | 4 | 4.222 | 0.176 | 4.17 | 0.407 | 9.65 | 0.444 | 10.51 | 30 | 6 | | 5 | 19.580 | 0.659 | 3.37 | 1.551 | 7.92 | 1.685 | 8.61 | 30 | 6 | | Negative Control | 0.164 | 0.099 | NA | 0.029 | NA | 0.103 | NA | 25 | 6 | | Positive Control | 4.798 | 0.254 | 5.30 | 0.300 | 6.24 | 0.393 | 8.19 | 25 | 6 | | Testing Site 3, Reagent Lot 85 | | | | | | | | | | | 1 | 0.096 | 0.031 | NA | 0.026 | NA | 0.040 | NA | 28 | 6 | | 2 | 0.948 | 0.066 | 6.92 | 0.251 | 26.48 | 0.259 | 27.36 | 30 | 6 | | 3 | 2.085 | 0.131 | 6.27 | 0.222 | 10.64 | 0.257 | 12.35 | 30 | 6 | | 4 | 4.865 | 0.251 | 5.17 | 0.421 | 8.66 | 0.491 | 10.08 | 30 | 6 | | 5 | 21.912 | 0.740 | 3.38 | 1.216 | 5.55 | 1.424 | 6.50 | 30 | 6 | | Negative Control | 0.174 | 0.074 | NA | 0.124 | NA | 0.145 | NA | 30 | 6 | | Positive Control | 4.934 | 0.287 | 5.82 | 0.224 | 4.54 | 0.364 | 7.38 | 30 | 6 | a Variability of the assay performance within day (from replicate to replicate). b Variability of the assay performance between days (from day to day). c Variability of the assay performance combining the effects of within day and between days. CV = Coefficient of variance NA = not applicable Note: 5 replicates per panel in 1 run per day for 6 days. 34 {26} 35 # Summary of Safety and Effectiveness The data from all 3 sites and from all 3 reagent lots were combined to achieve SD and percent CV for within run, between run, between testing site, between lot, and total. The precision estimates were derived from variance component analysis. The reproducibility results are presented in the following table. | Bayer ADVIA Centaur®® HBsAg Assay Reproducibility Between Testing Sites and Between Reagent Lots Estimates (Across All Reagent Lots and All Testing Sites) | | | | | | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Panel Member | Mean ADVIA Centaur® HBsAg Index Value | Within Run^{a} | | Between Run^{b} | | Between Testing Site^{c} | | Between Lot^{d} | | Total^{e} | | Number of Observations | | | | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | | | 1 | 0.09 | 0.015 | NA | 0.000 | NA | 0.001 | NA | 0.000 | NA | 0.015 | NA | 177 | | 2 | 0.70 | 0.088 | 12.44 | 0.066 | 9.37 | 0.153 | 21.77 | 0.000 | 0.00 | 0.188 | 26.77 | 180 | | 3 | 1.75 | 0.096 | 5.49 | 0.076 | 4.36 | 0.160 | 9.13 | 0.074 | 4.25 | 0.215 | 12.27 | 180 | | 4 | 4.29 | 0.155 | 3.62 | 0.124 | 2.90 | 0.232 | 5.41 | 0.226 | 5.28 | 0.380 | 8.87 | 180 | | 5 | 19.96 | 0.482 | 2.42 | 0.415 | 2.08 | 0.726 | 3.64 | 0.861 | 4.31 | 1.294 | 6.48 | 180 | | Negative Control | 0.13 | 0.055 | NA | 0.019 | NA | 0.059 | NA | 0.000 | NA | 0.083 | NA | 175 | | Positive Control | 4.63 | 0.177 | 3.83 | 0.112 | 2.42 | 0.212 | 4.57 | 0.060 | 1.30 | 0.304 | 6.56 | 175 | a Variability of the assay performance within day (all testing sites and reagent lots). b Variability of the assay performance between days (all testing sites and reagent lots). c Variability of the assay performance between testing sites (from testing site to testing site). d Variability of the assay performance between reagent lots (from reagent lot to reagent lot, across all testing sites). e Variability of the assay performance incorporating all testing sites, all reagent lots, and all days. CV = Coefficient of variance NA = Not applicable Note: 5 replicates per panel in 1 run per day for 6 days # Interpretation of Clinical Studies The following pattern of results interpretation was established on the basis of information collected from the clinical studies for the ADVIA Centaur® HBsAg assay: - Samples with an Index Value of less than 1.00 are considered nonreactive (negative) for HBsAg. - Samples with an Index Value of greater than or equal to 1.00 but less than or equal to 50 are considered initially reactive for HBsAg. Perform repeat testing in duplicate and/or supplemental testing on these samples. - Heparin anticoagulants have been shown to reduce the Index values in some samples near the assay cut off. High negative results (0.70 – 0.99 Index) obtained on samples collected with lithium or sodium heparin anticoagulant should be interpreted accordingly. Additional testing is required including other HBV markers and supplemental tests. - After repeat testing, if two of the three results are nonreactive (negative), the sample is considered negative for HBsAg. - After repeat testing, if at least two of the three results are reactive, the sample is considered repeat reactive for the presence of HBsAg. If a repeatedly reactive result is confirmed by supplemental tests, such as the ADVIA Centaur® HBsAg Confirmatory assay, the sample is positive for HBsAg. - If the sample is greater than 50, the specimen is positive for HBsAg by the {27} Summary of Safety and Effectiveness ADVIA Centaur® HBsAg assay. ## Device Failures and Replacements During the clinical studies there were no reported failures and/or replacements. ## XI. CONCLUSIONS DRAWN FROM THE STUDIES Multicenter studies were conducted in the US. The ADVIA Centaur® HBsAg assay performed with analytical sensitivity, analytical specificity, and positive agreement and negative agreement to reference methods, i.e., current commercially available FDA licensed or approved assays. - Hepatitis B virus classification using the prospective study population showed 31 unique reference marker patterns. The overall positive and negative percent agreement between the ADVIA Centaur® HBV method and the reference assay was reasonable in the high risk, signs and symptoms, and dialysis populations combined. - For the prenatal population, the overall positive and negative percent agreement between the ADVIA Centaur® HBV method and the reference assay was reasonable in all the trimesters tested. - In the HBV infected acute and chronic retrospective population, the overall positive percent agreement was acceptable. As all samples were reference method positive, there is no negative percent agreement calculation. - The ADVIA Centaur® HBsAg Confirmatory assay showed positive neutralization for 97.0 percent of samples that were repeatedly reactive by the ADVIA Centaur® HBsAg assay and 94.6 percent agreement with the reference assay for samples that were repeatedly reactive by the ADVIA Centaur® HBsAg assay for the high risk, signs and symptoms, dialysis, and prenatal populations combined. - The ability of the ADVIA Centaur® HBsAg assay to detect HBV infection shortly after initial HBV infection was demonstrated with the seroconversion panel evaluation. When the ADVIA Centaur® HBsAg result was compared to the reference assay results the detection of the first reactive specimen was similar to that of the reference method. - Precision and reproducibility of the ADVIA Centaur® HBsAg assay was acceptable with minor variability from run to run, day to day, or reagent lot to reagent lot. - Paired Matrix Study results supported the use of human serum, EDTA plasma, and lithium heparin plasma specimens for testing in the ADVIA Centaur® HBsAg assay. The use of heparin anticoagulants required an adjustment of the assay cutoff for these specimens. The results of the non-clinical and clinical studies indicate that the ADVIA Centaur® HBsAg and ADVIA Centaur® HBsAg Confirmatory assays can be used safely and 36 {28} Summary of Safety and Effectiveness effectively for the qualitative *in vitro* determination of HBsAg in human serum and plasma. The ADVIA Centaur® HBsAg and ADVIA Centaur® HBsAg Confirmatory assays may be used with other HBV serological markers to define the clinical status of patients known to be infected with HBV or with other HBV, HAV, and HCV assays to form a panel for the diagnosis of patients presenting with symptoms of acute or chronic viral hepatitis. In addition, the ADVIA Centaur® HBsAg and ADVIA Centaur® HBsAg Confirmatory assays can be used safely and effectively for the identification of neonates who are at risk for HBV infection during the perinatal period. ## RISK BENEFIT ANALYSIS As a diagnostic test, the ADVIA Centaur® HBsAg Assay involves removal of blood from an individual for testing purposes. This test presents no more of a safety hazard to an individual than is presented to an individual who is having their blood drawn for any other diagnostic evaluations. The potential risks encountered with *in vitro* diagnostic tests are not unusual in the clinical laboratory setting. Appropriate warnings for these risks are contained in the labeling and package inserts for these devices. Standard good laboratory practices are considered sufficient to mitigate the risks to the individual tested. The rate of false positivity and false negativity for the ADVIA Centaur® are within acceptable limits compared to the reference assay. It has been shown that the device has no demonstrable cross-reactivity with the viruses or organisms tested that may cause clinical hepatitis. ## SAFETY Based on the results of the preclinical and clinical laboratory studies, the ADVIA Centaur® HBsAg assays, when used according to the provided directions and in conjunction with other serological and clinical information, should be safe and pose minimal risk to the patient due to false test results. ## EFFECTIVENESS The effectiveness of the ADVIA Centaur® HBsAg has been demonstrated for use in determining if HBsAg is present in an individual’s serum or plasma. A reasonable determination of effectiveness of the ADVIA Centaur® HBsAg assays for aiding in the diagnosis of acute and chronic HBV infection and prenatal screening has been demonstrated. ## XII. PANEL RECOMMENDATION In accordance with the provisions section 515(c)(2) of the act as amended by the Safe Medical Devices Act of 1990, this PMA was not referred to the Microbiology Devices Advisory Panel an FDA advisory committee, for review and recommendation because the information in the PMA substantially duplicated information previously reviewed by this Panel. 37 {29} Summary of Safety and Effectiveness ## XIII. CDRH DECISION FDA issued an approval order on May 26, 2005. The applicant’s manufacturing facility was inspected on May 11, 2004 (New York) and April 28, 2004 (Massachusetts) and was found to be in compliance with the Quality System Regulation (21 CFR 820). ## XIV. APPROVAL SPECIFICATIONS Directions for Use: See the labeling. Hazards to Health from Use of the Device: See Indications, Contraindications, Warnings, Precautions and Adverse Events in the labeling. Postapproval Requirements and Restrictions: See approval order. 38
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