ADVIA Centaur Anti-HBe2 (aHBe2) assay

P200017 · Siemens Healthcare Diagnostics, Inc. · SEI · Jul 14, 2021 · Microbiology

Device Facts

Record IDP200017
Device NameADVIA Centaur Anti-HBe2 (aHBe2) assay
ApplicantSiemens Healthcare Diagnostics, Inc.
Product CodeSEI · Microbiology
Decision DateJul 14, 2021
DecisionAPRL
Regulation21 CFR 866.3173
Device ClassClass 2
AttributesPediatric

Intended Use

The ADVIA Centaur® Anti-HBe2 (aHBe2) assay is an in vitro diagnostic immunoassay for the qualitative detection of antibodies to the e antigen of the hepatitis B virus (HBV) in human pediatric (2–21 years old) and adult serum, EDTA plasma, or lithium heparin plasma using the ADVIA Centaur systems (XP/XPT/CP). Assay results, in conjunction with other laboratory results and clinical information may be used as an aid in the diagnosis of hepatitis B virus (HBV) infection in patients with signs or symptoms of hepatitis B infection, or with risk factors for HBV infection, or with known HBV infection. Results of the assay, in conjunction with other diagnostic information, may be used to aid in determining HBV seroconversion. This assay is not intended for screening donors of blood or blood products or human cells, tissues, and cellular and tissue-based products (HCT/Ps).

Device Story

ADVIA Centaur Anti-HBe2 is a fully automated 2-step competitive chemiluminescent immunoassay. Input: human serum, EDTA plasma, or lithium heparin plasma. Process: sample incubated with recombinant HBeAg; remaining HBeAg binds to solid-phase microparticles coated with anti-HBe monoclonal antibody and acridinium ester-labeled anti-HBe monoclonal antibody. Output: relative light units (RLUs) inversely proportional to anti-HBe concentration, reported as reactive/nonreactive Index values. Used in clinical laboratories by professional staff. Output aids clinicians in diagnosing HBV infection, monitoring disease progression, and managing patients undergoing antiviral therapy. Benefits include accurate detection of HBV seroconversion and improved patient management.

Clinical Evidence

Multisite clinical study evaluated 1814 prospective samples (pediatric and adult) against an FDA-approved reference assay. Performance across ADVIA Centaur XP, XPT, and CP systems showed high agreement. Overall PPA ranged 92.0%–97.6% and NPA 99.0%–99.1% depending on the system. Pediatric performance (N=180) showed 100% PPA and 99.1% NPA. Study included pregnant women (N=228) with acceptable performance across trimesters. No clinical cross-reactivity observed with 314 samples from 33 unrelated medical conditions.

Technological Characteristics

Competitive chemiluminescent immunoassay. Reagents: recombinant HBeAg, paramagnetic microparticles coated with mouse monoclonal anti-HBe, acridinium ester-labeled mouse monoclonal anti-HBe. Instrumentation: ADVIA Centaur XP, XPT, and CP automated analyzers. Connectivity: networked laboratory systems. Software: automated data reduction and result reporting. Calibration: two-point, lot-specific Master Curve.

Indications for Use

Indicated for qualitative detection of anti-HBe antibodies in human serum, EDTA plasma, or lithium heparin plasma from pediatric (2–21 years) and adult patients. Used as an aid in diagnosis of HBV infection in patients with signs/symptoms, risk factors, or known HBV infection, and to aid in determining HBV seroconversion. Not for blood/tissue donor screening.

Regulatory Classification

Identification

A hepatitis B virus (HBV) antibody assay is identified as an in vitro diagnostic device intended for prescription use in the detection of antibodies to HBV in human serum, plasma, or other matrices, and as a device that aids in the diagnosis of HBV infection in persons with signs and symptoms of hepatitis and in persons at risk for hepatitis B infection. Results from assays may be qualitative or quantitative, such as quantitative anti-HBs. In addition, results from an anti-HBc IgM (IgM antibodies to core antigen) assay indicating the presence of anti-HBc IgM are indicative of recent HBV infection. Anti-HBs (antibodies to surface antigen) assay results may be used as an aid in the determination of susceptibility to HBV infection in individuals prior to or following HBV vaccination or when vaccination status is unknown. The assay is not intended for screening of blood, plasma, cells, or tissue donors. The assay is intended as an aid in diagnosis in conjunction with clinical findings and other diagnostic procedures.

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) When appropriate, performance characteristics of the assay have not been established in populations of immunocompromised or immunosuppressed patients or other special populations where assay performance may be affected. (B) Detection of HBV antibodies to a single viral antigen indicates a present or past infection with hepatitis B virus, but does not differentiate between acute, chronic, or resolved infection. (C) The specimen types for which the device has been cleared, and that use of the assay with specimen types other than those specifically cleared for this device may result in inaccurate assay results. (D) 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. (E) A non-reactive assay result may occur early during acute infection, prior to development of a host antibody response to infection, or when analyte levels are below the limit of detection of the assay. (F) Results obtained with this assay may not be used interchangeably with results obtained with a different manufacturer's assay. (v) For devices intended for the quantitative detection of HBV antibodies (anti-HBs), in addition to the special controls listed in paragraphs (b)(1) and (2) of this section, labeling required under § 809.10(b) of this chapter must include: (A) The assay calibrators' traceability to a standardized reference material that FDA has determined is appropriate ( *e.g.,* a recognized consensus standard) and the limit of blank (LoB), limit of detection (LoD), limit of quantitation (LoQ), linearity, and precision to define the analytical measuring interval.(B) Performance results of the analytical sensitivity study testing a standardized reference material that FDA has determined is appropriate ( *e.g.,* a recognized consensus standard).(2) Design verification and validation must include the following: (i) Detailed device description, including all parts that make up the device, ancillary reagents required but not provided, an explanation of the device methodology, and design of the antigen(s) and capture antibody(ies) sequences, rationale for the selected epitope(s), degree of amino acid sequence conservation of the target, and the design and composition of all primary, secondary and subsequent standards used for calibration. (ii) Documentation and characterization ( *e.g.,* supplier, determination of identity, and stability) of all critical reagents (including description of the antigen(s) and capture antibody(ies)), and protocols for maintaining product integrity throughout its labeled shelf life.(iii) 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. (iv) 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. (v) 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. (vi) All stability protocols, including acceptance criteria. (vii) When applicable, analytical sensitivity of the assay that is the same or better than that of other cleared or approved assays. (viii) Analytical performance studies and results for determining 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, specimen stability, reagent stability, and cross-genotype antibody detection sensitivity, when appropriate. (ix) For devices intended for the detection of antibodies for which a standardized reference material (that FDA has determined is appropriate) is available, the analytical sensitivity study and results testing the standardized reference material. Detailed documentation of that study and its results must be provided, including the study protocol, study report, testing results, and all statistical analyses. (x) 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. (xi) Detailed documentation of clinical performance testing from a clinical study with an appropriate number of HBV reactive and non-reactive samples in applicable risk categories and conducted in the appropriate settings by the intended users. Performance must be analyzed relative to an FDA cleared or approved HBV antibody assay or a comparator that FDA has determined is appropriate. 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. (3) For any HBV antibody assay intended for quantitative detection of anti-HBV antibodies, the following special controls, in addition to those special controls listed in paragraphs (b)(1) and (2) of this section, also apply: (i) Detailed documentation of the metrological calibration traceability hierarchy to a standardized reference material that FDA has determined is appropriate. (ii) Detailed documentation of the following analytical performance studies conducted, as appropriate to the technology, specimen types tested, and intended use of the device, including upper and lower limits of quantitation (UloQ and LloQ, respectively), linearity using clinical samples, and an accuracy study using the recognized international standard material.

Related Devices

Submission Summary (Full Text)

{0} # SUMMARY OF SAFETY AND EFFECTIVENESS DATA (SSED) ## I. GENERAL INFORMATION Device Generic Name: Antibody to Hepatitis B e Antigen Assay (Anti-HBe) Device Trade Name: ADVIA Centaur® Anti-HBe2 (aHBe2) assay Device Product Code: LOM Applicants Name and Address: Siemens Healthcare Diagnostics Inc. 511 Benedict Ave Tarrytown, NY 10591 Date of Panel Recommendation: None Premarket Approval Application (PMA) Number: P200017 Date of FDA Notice of Approval: July 14, 2021 ## II. INDICATIONS FOR USE The ADVIA Centaur® Anti-HBe2 (aHBe2) assay is an in vitro diagnostic immunoassay for the qualitative detection of antibodies to the e antigen of the hepatitis B virus (HBV) in human pediatric (2–21 years old) and adult serum, EDTA plasma, or lithium heparin plasma using the ADVIA Centaur systems (XP/XPT/CP). Assay results, in conjunction with other laboratory results and clinical information may be used as an aid in the diagnosis of hepatitis B virus (HBV) infection in patients with signs or symptoms of hepatitis B infection, or with risk factors for HBV infection, or with known HBV infection. Results of the assay, in conjunction with other diagnostic information, may be used to aid in determining HBV seroconversion. This assay is not intended for screening donors of blood or blood products or human cells, tissues, and cellular and tissue-based products (HCT/Ps). ## III. CONTRAINDICATIONS There are no known contraindications. PMA P200017: FDA Summary of Safety and Effectiveness Data Page 1 of 39 {1} PMA P200017: FDA Summary of Safety and Effectiveness Data Page 2 of 39 # IV. WARNINGS AND PRECAUTIONS The warning and precautions can be found in the ADVIA Centaur® Anti-HBe2 (aHBe2) assay package insert. - For in vitro diagnostic use. - For Professional Use. - For Prescription Use Only. - Warning - May cause an allergic skin reaction. Wear protective gloves/protective clothing/eye protection/face protection. IF ON SKIN: Wash with plenty of soap and water. If skin irritation or rash occurs: Get medical advice/attention. Take off contaminated clothing and wash it before reuse. - Caution Potential Biohazard - Contains human source material. Each donation of human blood or blood component negative for antibodies to HBe antigen was tested by FDA-approved methods for the presence of antibodies to human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2), as well as for hepatitis B surface antigen (HBsAg) and antibody to hepatitis C virus (HCV). The test results were negative (not repeatedly reactive). No test offers complete assurance that these or other infectious agents are absent; this material should be handled using good laboratory practices and universal precautions. Calibrators and controls contain processed human plasma that is nonreactive for HIV and HCV but is reactive for antibodies to HBe antigen. The units were treated with a BPL-UV inactivation procedure; 10 however, all products manufactured using human source material should be handled as potentially infectious. - Caution - This device contains material of animal origin and should be handled as a potential carrier and transmitter of disease. # V. DEVICE DESCRIPTION ## Assay Principle and Format: The ADVIA Centaur aHBe2 assay is a fully automated 2-step competitive immunoassay using chemiluminescent technology The Ancillary well reagent contains recombinant HBeAg. The Solid Phase contains microparticles coated with aHBe monoclonal antibody. The Lite Reagent contains aHBe monoclonal antibody labeled with acridinium ester. The sample is incubated with the Ancillary Well Reagent. During the incubation period, aHBe in the sample binds with recombinant HBeAg. The Solid Phase and Lite Reagent are added next and {2} bind any recombinant HBeAg not already bound by sample. After a wash step, acid and base are dispensed to initiate the chemiluminescent reaction. An inverse relationship exists between the amount of aHBe activity present in the patient sample and the amount of relative light units (RLUs) detected by the system. A result of reactive or nonreactive is determined according to a cut-off of 1.00 Index established with the calibrators. The ADVIA Centaur XP system automatically performs the following actions: 1. Dispenses 75 μL of sample into a cuvette. 2. Dispenses 75 μL of Ancillary Well Reagent and incubates the mixture for 29 minutes at 37°C. 3. Dispenses 150 μL of Solid Phase and 75 μL of Lite Reagent, and incubates the mixture for 18 minutes at 37°C. 4. Separates, aspirates, then washes the cuvette with ADVIA Centaur Wash 5. Dispenses 300 μL each of ADVIA Centaur Acid Reagent and ADVIA Centaur Base Reagent to initiate the chemiluminescent reaction. 6. Reports results. ## Calibration: The ADVIA Centaur aHBe2 assay utilizes two-point calibration using the two calibrators (Calibrator L, Calibrator H) provided with each kit. There are two levels, the low calibrator (Calibrator L (~ 0.5 Index)) and the high calibrator (Calibrator H (~ 1.5 Index)). When used with the ADVIA Centaur aHBe2 reagent, there is an emission of light which can be measured by the ADVIA Centaur luminometer. The ADVIA Centaur aHBe2 calibrators are value assigned over multiple runs on the ADVIA Centaur instruments using a specific lot of reagent and against internal standards. Results are reported in Index value. The 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. Calibration frequency- The two calibrators in the kit are run when one or more of the following conditions exist: - At the end of the 21-day calibration interval. - When changing lot numbers of primary reagent packs. - When indicated by quality control results. - After major maintenance or service, as indicated by quality control results. 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 Index value of the sample or control is read off the Master Curve. Individuals whose samples read at or above an Index of 1.0 are considered to be reactive for aHBe while those below 1.00 are non-reactive. PMA P200017: FDA Summary of Safety and Effectiveness Data Page 3 of 39 {3} PMA P200017: FDA Summary of Safety and Effectiveness Data Page 4 of 39 # Controls: ADVIA Centaur Anti-HBe2 QC set contains Negative control (2 vials with 10 mL) and Positive control (2 vials with 10 mL). They have different concentrations of anti-HBe antibodies and the Index (Negative control (~0.3 Index) and Positive control (~2.00 Index)) for each control is indicated in a Control Expected Value card enclosed with the kit. The performance of the ADVIA Centaur aHBe2 assay is monitored by the use of Centaur aHBe2 Quality Controls at least once during each day when samples are analyzed or after a successful calibration. # Interpretation of Results: The sample results from the ADVIA Centaur aHBe2 assay are in Index Values and they are reported by the system as 'Non-reactive' or 'Reactive.' The final interpretation of the results is done as shown below in Table 1. Table 1. Interpretation of Results for the ADIVIA Centaur aHBe2 Assay | Initial Test Results (C/S) | Retest Zone | Final Interpretation | | --- | --- | --- | | < 0.80 | Not Required | Non-Reactive (Negative) | | ≥ 0.80 and < 1.20 | Retest in duplicate | Sample is non-reactive and presumed negative for anti-HBe if 2 of 3 results are <1.00. Sample is reactive and presumed positive for anti-HBe if 2 of 3 results are ≥ 1.00. | | ≥ 1.20 | Not Required | Reactive (Positive) | # Components of the ADVIA Centaur® Anti-HBe2 (aHBe2) Assay ## 1. Reagent Kit: Each ADVIA Centaur Anti-HBe2 kit contains a ReadyPack for 50 tests and one vial each of two calibrators (Calibrator L and Calibrator H). The calibrators provided in the kit are matched to the ReadyPack primary reagent. The kit also provides ADVIA Centaur aHBe2 Master Curve card and aHBe2 CAL calibrator assigned value card and barcode labels. ADVIA Centaur Anti-HBe2 ReadyPack – Number of Tests – 50 - **Lite reagent: 3.8 mL/reagent pack.** Mouse monoclonal anti-HBe labeled with an acridinium ester in buffer, bovine serum albumin (BSA); mouse IgG; surfactant; preservative Proclin 950 1.5 mL/L. - **Solid phase: 7.5 mL/reagent pack** Paramagnetic microparticle suspension coated with mouse monoclonal aHBe in buffer; BSA, surfactant; sodium azide (&lt; 0.1%). - **Ancillary reagent: 3.8 mL/reagent pack** {4} Recombinant HBe antigen in buffer; BSA; surfactant; preservative Proclin 950 1.5 mL/L. ADVIA Centaur Anti-HBe2 Calibrator Processed human plasma negative and positive for aHBe; sodium azide (&lt; 0.1%) - aHBe2 CAL low calibrator: 2.0 mL/vial - aHBe2 CAL high calibrator: 2.0 mL/vial ## Additional materials required but purchased separately ### 2. ADVIA Centaur aHBe2 Quality Control Kit: Negative Control: 2 x 10.0 mL, Level 1 Positive Control: 2 x 10.0 mL, Level 2 Quality control lot-specific value sheet and barcode labels. ### 3. ADVIA Centaur Probe Wash 3: 50.0 mL/pack Sodium hypochlorite (0.5%); sodium hydroxide (&lt;0.5%), pH 11.0 ### 4. ADVIA Centaur Wash 1: 1500 mL/pack Phosphate buffered saline; sodium azide (&lt; 0.1%); surfactant ### 5. ADVIA Centaur Wash 1: 2500 mL/Pack Phosphate buffered saline; sodium azide (&lt; 0.1%); surfactant ### 6. Instrumentation and Software: The ADVIA Centaur CP and ADVIA Centaur XP systems were cleared under k041133 and k032525. The ADVIA Centaur XPT is an upgraded model and instrument family member of the XP system cleared under k141999. The Siemens ADVIA Centaur XP, ADVIA Centaur XPT and ADVIA Centaur CP System are automated immunoassay analyzers designed to perform in vitro diagnostic immunochemical assay analysis on clinical specimens and intended for professional use in a laboratory environment only. The system menu includes endocrine, anemia, allergy, reproductive, cardiovascular, oncology, adrenal, bone metabolism, therapeutic drug, and infectious disease assays. All assays are based on chemiluminescent technology. The ADVIA Centaur CP, ADVIA Centaur XP and ADVIA Centaur XPT Systems perform the following functions: - Aspirates and dispenses samples - Performs dilutions - Adds reagents - Incubates reaction vessels - Separates solid and liquid wastes - Measures photon emissions PMA P200017: FDA Summary of Safety and Effectiveness Data Page 5 of 39 {5} - Performs data reduction - Collects and maintains patient demographics and results. The ADVIA Centaur XP, XPT and CP systems are automated and perform different functions including loading samples, primary and ancillary reagents, and supplies, or emptying waste without pausing or interrupting test processing. The ADVIA Centaur XP, XPT and CP computers control system functions and processes data. The applications module is equipped with a CD-RW drive for software updates and for data storage. The ADVIA Centaur System software are the set of computer instructions that interprets system and assay information, calculates results, and provides the interface for controlling the system hardware. The ADVIA Centaur XP software provides user interface workstation to control the operation of the system. The workspace has a system view offering access to system functions and an applications view offering access to a set of applications including the ADVIA QC analyzer, the RealTime Solutions service (e.g., remote troubleshooting and repair) and the online documentation browser. Off-The-Shelf software components are integrated into the overall design of the ADVIA Centaur CP, ADVIA Centaur XP and ADVIA Centaur XPT software and as a result do not function outside of the software. The end user has no direct interaction with the OTS software components but instead with the overall design of the ADVIA Centaur CP, ADVIA Centaur XP and ADVIA Centaur XPT software. Additional details can be found in the ADVIA Centaur XP, XPT or CP Operating Instructions Manual. ## VI. ALTERNATIVE PRACTICES AND PROCEDURES There are several other alternatives for the determination of HBV infection and its disease stage. Detection of anti-HBe in patients who may be infected with the hepatitis B virus may also be accomplished with any commercially available FDA approved serological tests. This assay is one of several hepatitis marker assays that are often used together in conjunction with clinical assessment and other laboratory test results in the diagnosis of the HBV infection. A patient should fully discuss these alternatives with his/her physician to select the method that best meets expectations and lifestyle. ## VII. MARKETING HISTORY The ADVIA Centaur aHBe2 assay has not been marketed in the United States or any foreign country. PMA P200017: FDA Summary of Safety and Effectiveness Data Page 6 of 39 {6} # VIII. POTENTIAL ADVERSE EFFECTS OF THE DEVICE ON HEALTH The potential adverse effects associated with the use of this device are as follows. Failure of the product to perform as indicated or human error in use of the product may lead to a false result. Anti-HBe antibody is an intermediate or long-term risk analyte; repeatedly erroneous false positive or false negative anti-HBe results could lead to inappropriate initiation or cessation of antiviral therapy. When used according to the instructions in the package insert, there are no known direct adverse effects of this device on the health of the user. No specific adverse effects occurred during the conduct of clinical studies. # IX. SUMMARY OF NON-CLINICAL STUDIES The software versions used for the studies described below were ADVIA Centaur CP (v6.4), ADVIA Centaur XP (v7.4.0.4) and ADVIA Centaur XPT (v1.5.1). # Cut-Off Determination A set of 2111 samples that had been previously tested and classified as Positive (204) or Negative (1907) using the FDA-approved Reference Assay (Anti-HBe Assay) were tested with the ADVIA Centaur Anti-HBe2 assay on ADVIA Centaur XP system to determine the Cut-Off. For each sample, sensitivity (TP rate) and specificity (TN rate) versus the Reference Assay was calculated, using the individual RLU of ADVIA Centaur Anti-HBe2 result as a tentative Cut-Off. The optimal ADVIA Centaur aHBe2 Cut-Off was obtained considering the best results of the area under the ROC (Receiver Operating Characteristic) curve in terms of discrimination between positive and negative samples according to the reference method and considering the criterion of maximize the relative sensitivity and specificity. The ADVIA Centaur aHBe2 assay Cut-Off (1.00 Index) was established at 214986 RLU, because it showed a good Sensitivity without compromising the Specificity. The percent positive agreement and percent negative agreement with the reference assay used to determine the cut-off were $\geq 95.0\%$. # Precision The precision study for the ADVIA Centaur Anti-HBe2 assay was conducted in various sample matrices in a 20-day precision protocol using three lots of the reagents on ADVIA Centaur XP, XPT and CP instruments. Two controls (Positive &amp; Negative) and four specimens in three matrices (serum, EDTA plasma, and lithium heparin plasma) were used to measure the precision of the assay at different dose levels. The specimens were assayed in duplicate per run, two runs per day for 20 days (N = 80 for each sample). PMA P200017: FDA Summary of Safety and Effectiveness Data Page 7 of 39 {7} Four samples for each matrix was selected at different nominal values: sample 1, 0.50-0.79 (high non-reactive), sample 2, 0.80-1.20 (Cut-Off), sample 3, 1.21-1.70 (Reactive) and sample 4, $&gt;1.70$ (High Reactive). The results in the following tables are with one lot of the ADVIA Centaur Anti-HBe2 assay on XP instrument. Calculations for within-run, between-run, between-day, and total precision were performed according to CLSI Document EP05-A3. The repeatability of the combined three lots of ADVIA Centaur Anti-HBe2 assay ranged from $0.8\%$ to $4.9\%$ . The within-laboratory $\% \mathrm{CV}$ of the combined three lots ranged from $1.3\%$ to $9.6\%$ . The results obtained with three lots of Reagent on ADVIA Centaur XP (Table 2A), XPT (Table 2B) and CP (Table 2C) instruments for repeatability and within-laboratory, indicating only minor variability in the components evaluated. Table 2A. Centaur XP - Summary of Precision Results for the ADVIA Centaur Anti-HBe2 Assay | Sample Description | Mean (Index) | Within-Run | | Between-Run | | Between-Day | | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | \( SD^a \) | \( CV^b \)% | SD | CV % | SD | CV % | SD | CV % | | EDTA plasma 1 | 0.71 | 0.01 | 1.7% | 0.01 | 0.8% | 0.01 | 1.8% | 0.02 | 2.6% | | EDTA plasma 2 | 1.06 | 0.01 | 1.2% | 0.01 | 1.1% | 0.01 | 1.2% | 0.02 | 2.0% | | EDTA plasma 3 | 1.41 | 0.02 | 1.7% | 0.02 | 1.2% | 0.01 | 0.5% | 0.03 | 2.2% | | EDTA plasma 4 | 1.91 | 0.02 | 1.0% | 0.03 | 1.3% | 0.00 | 0.0% | 0.03 | 1.6% | | Lithium Heparin plasma 1 | 0.61 | 0.01 | 2.1% | 0.01 | 2.3% | 0.01 | 1.5% | 0.02 | 3.4% | | Lithium Heparin plasma 2 | 1.02 | 0.02 | 1.5% | 0.02 | 1.6% | 0.01 | 0.6% | 0.02 | 2.3% | | Lithium Heparin plasma 3 | 1.43 | 0.02 | 1.3% | 0.01 | 0.9% | 0.01 | 0.9% | 0.03 | 1.8% | | Lithium Heparin plasma 4 | 1.96 | 0.03 | 1.5% | 0.02 | 1.2% | 0.00 | 0.0% | 0.04 | 1.9% | | Serum 1 | 0.54 | 0.01 | 1.9% | 0.01 | 1.5% | 0.01 | 1.8% | 0.02 | 3.0% | | Serum 2 | 0.84 | 0.01 | 1.6% | 0.03 | 3.5% | 0.00 | 0.0% | 0.03 | 3.9% | | Serum 3 | 1.44 | 0.02 | 1.4% | 0.02 | 1.3% | 0.01 | 0.6% | 0.03 | 2.0% | | Serum 4 | 2.00 | 0.04 | 1.7% | 0.02 | 1.0% | 0.02 | 0.9% | 0.04 | 2.2% | | Negative Control | 0.28 | 0.01 | 3.1% | 0.01 | 3.5% | 0.01 | 3.8% | 0.02 | 6.0% | | Positive Control | 1.96 | 0.02 | 1.2% | 0.01 | 0.6% | 0.01 | 0.7% | 0.03 | 1.5% | ${}^{a}$ Standard deviation of mean concentration (Index) b Coefficient of variation PMA P200017: FDA Summary of Safety and Effectiveness Data Page 8 of 39 {8} Table 2B. Centaur XPT - Summary of Precision Results for the ADVIA Centaur Anti-HBe2 Assay | Sample Description | Mean (Index) | Within-Run | | Between-Run | | Between-Day | | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | \( SD^a \) | \( CV^b \)% | SD | CV % | SD | CV % | SD | CV % | | EDTA plasma 1 | 0.74 | 0.02 | 2.9% | 0.01 | 1.3% | 0.03 | 4.3% | 0.04 | 5.3% | | EDTA plasma 2 | 1.07 | 0.03 | 3.1% | 0.03 | 3.2% | 0.03 | 2.8% | 0.06 | 5.3% | | EDTA plasma 3 | 1.45 | 0.03 | 2.1% | 0.03 | 1.7% | 0.03 | 2.1% | 0.05 | 3.4% | | EDTA plasma 4 | 1.99 | 0.06 | 2.9% | 0.03 | 1.6% | 0.03 | 1.7% | 0.07 | 3.7% | | Lithium Heparin plasma 1 | 0.63 | 0.02 | 3.2% | 0.02 | 3.6% | 0.02 | 2.5% | 0.03 | 5.4% | | Lithium Heparin plasma 2 | 1.06 | 0.02 | 2.1% | 0.03 | 2.4% | 0.03 | 3.0% | 0.05 | 4.4% | | Lithium Heparin plasma 3 | 1.47 | 0.03 | 2.0% | 0.03 | 1.9% | 0.04 | 2.6% | 0.06 | 3.8% | | Lithium Heparin plasma 4 | 2.04 | 0.07 | 3.4% | 0.01 | 0.6% | 0.04 | 1.9% | 0.08 | 4.0% | | Serum 1 | 0.55 | 0.02 | 2.8% | 0.02 | 4.2% | 0.02 | 4.4% | 0.04 | 6.8% | | Serum 2 | 0.84 | 0.03 | 3.0% | 0.03 | 3.3% | 0.02 | 2.3% | 0.04 | 5.0% | | Serum 3 | 1.47 | 0.03 | 1.8% | 0.04 | 2.7% | 0.02 | 1.3% | 0.05 | 3.5% | | Serum 4 | 2.08 | 0.07 | 3.2% | 0.02 | 1.1% | 0.04 | 2.0% | 0.08 | 4.0% | | Negative Control | 0.28 | 0.01 | 4.7% | 0.01 | 5.1% | 0.02 | 6.6% | 0.03 | 9.6% | | Positive Control | 2.04 | 0.05 | 2.4% | 0.03 | 1.6% | 0.04 | 2.0% | 0.07 | 3.5% | ${}^{a}$ Standard deviation of mean concentration (Index) b Coefficient of variation Table 2C. ADVIA Centaur CP - Summary of Precision Results for the ADVIA Centaur Anti-HBe2 Assay | Sample Description | Mean (Index) | Within-Run | | Between-Run | | Between-Day | | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | \( SD^a \) | \( CV^b \)% | SD | CV % | SD | CV % | SD | CV % | | EDTA plasma 1 | 0.72 | 0.01 | 1.2% | 0.01 | 1.9% | 0.01 | 0.9% | 0.02 | 2.4% | | EDTA plasma 2 | 1.06 | 0.02 | 2.1% | 0.01 | 1.2% | 0.01 | 0.7% | 0.03 | 2.5% | | EDTA plasma 3 | 1.44 | 0.02 | 1.2% | 0.02 | 1.4% | 0.01 | 0.9% | 0.03 | 2.1% | | EDTA plasma 4 | 1.97 | 0.02 | 1.1% | 0.03 | 1.3% | 0.04 | 1.9% | 0.05 | 2.5% | | Lithium Heparin plasma 1 | 0.62 | 0.01 | 1.3% | 0.01 | 1.7% | 0.01 | 2.1% | 0.02 | 3.0% | PMA P200017: FDA Summary of Safety and Effectiveness Data {9} # System Reproducibility The system reproducibility of the ADVIA Centaur Anti-HBe2 assay was evaluated on ADVIA Centaur XP, XPT and CP systems at three external US sites. Each site tested at least two of the three reagent lots. A four-member serum panel was assayed in replicates of four with two runs per day, over five days for each lot $(N = 240)$ for each sample. The system reproducibility was determined in accordance with CLSI Document EP05-A3 Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline—Third Edition. The $\%$ CV for reproducibility for the three sites are ranged between $2.3\% -10.0\%$ . The results for ADVIA Centaur XP, XPT, and CP are summarized below in the Tables 3A, 3B and3C. Table 3A. ADVIA Centaur XP - Summary of Anti-HBe2 Reproducibility Results | Sample | Mean (Index) | Within-Run Repeatability | | Between-Run | | Between-Day | | Between-Lot | | Between-Site | | Reproducibility | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | \( SD^a \) | \( CV^b \)(%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | | Serum 1 | 0.61 | 0.01 | 2.4 | 0.01 | 1.6 | 0.00 | 0.5 | 0.05 | 7.8 | 0.00 | 0.0 | 0.05 | 8.3 | | Serum 2 | 0.95 | 0.02 | 1.9 | 0.02 | 1.7 | 0.01 | 1.2 | 0.06 | 6.1 | 0.00 | 0.0 | 0.06 | 6.7 | | Serum 3 | 1.46 | 0.02 | 1.6 | 0.02 | 1.3 | 0.01 | 0.6 | 0.01 | 0.8 | 0.00 | 0.0 | 0.03 | 2.3 | | Serum 4 | 2.08 | 0.03 | 1.5 | 0.01 | 0.6 | 0.02 | 0.9 | 0.05 | 2.6 | 0.00 | 0.0 | 0.07 | 3.2 | | Negative Control | 0.28 | 0.01 | 3.7 | 0.01 | 2.3 | 0.00 | 1.5 | 0.02 | 6.8 | 0.00 | 0.0 | 0.02 | 8.2 | | Positive Control | 1.99 | 0.03 | 1.6 | 0.02 | 0.8 | 0.01 | 0.5 | 0.03 | 1.3 | 0.00 | 0.0 | 0.05 | 2.3 | ${}^{a}$ Standard deviation of mean concentration (Index) ${}^{\mathrm{b}}$ Coefficient of variation PMA P200017: FDA Summary of Safety and Effectiveness Data {10} \mathrm{b} Coefficient of variation Table 3B. ADVIA Centaur XPT - Summary of Anti-HBe2 Reproducibility Results | Sample | Mean (Index) | Within-Run Repeatability | | Between-Run | | Between-Day | | Between-Lot | | Between-Site | | Reproducibility | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | \( SD^a \) | \( CV^b \)(%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | | Serum 1 | 0.60 | 0.01 | 2.3 | 0.01 | 1.2 | 0.01 | 1.0 | 0.05 | 8.3 | 0.00 | 0.0 | 0.05 | 8.8 | | Serum 2 | 0.93 | 0.02 | 1.6 | 0.02 | 1.7 | 0.00 | 0.0 | 0.05 | 5.9 | 0.00 | 0.0 | 0.06 | 6.4 | | Serum 3 | 1.44 | 0.02 | 1.7 | 0.02 | 1.1 | 0.00 | 0.2 | 0.01 | 0.4 | 0.02 | 1.4 | 0.04 | 2.5 | | Serum 4 | 2.01 | 0.03 | 1.4 | 0.01 | 0.3 | 0.01 | 0.5 | 0.03 | 1.4 | 0.05 | 2.3 | 0.06 | 3.1 | | Negative Control | 0.28 | 0.01 | 4.0 | 0.01 | 2.3 | 0.00 | 0.0 | 0.01 | 2.4 | 0.01 | 2.4 | 0.02 | 5.7 | | Positive Control | 1.96 | 0.03 | 1.3 | 0.02 | 0.9 | 0.00 | 0.0 | 0.03 | 1.5 | 0.03 | 1.7 | 0.05 | 2.7 | ${}^{a}$ Standard deviation of mean concentration (Index) b Coefficient of variation Table 3C. ADVIA Centaur CP - Summary of Anti-HBe2 Reproducibility Results | Sample | Mean (Index) | Within-Run Repeatability | | Between-Run | | Between-Day | | Between-Lot | | Between-Site | | Reproducibility | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | \( SD^a \) | \( CV^b \)(%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | | Serum 1 | 0.60 | 0.01 | 2.3 | 0.01 | 2.2 | 0.01 | 0.8 | 0.05 | 8.2 | 0.02 | 4.1 | 0.06 | 9.7 | | Serum 2 | 0.95 | 0.02 | 2.5 | 0.03 | 3.3 | 0.00 | 0.0 | 0.05 | 5.4 | 0.00 | 0.0 | 0.06 | 6.8 | | Serum 3 | 1.46 | 0.03 | 2.1 | 0.03 | 2.0 | 0.00 | 0.9 | 0.01 | 0.4 | 0.02 | 1.5 | 0.05 | 3.3 | | Serum 4 | 2.07 | 0.06 | 3.0 | 0.02 | 0.8 | 0.02 | 1.1 | 0.04 | 1.7 | 0.02 | 0.8 | 0.08 | 3.8 | | Negative Control | 0.28 | 0.01 | 5.2 | 0.01 | 2.7 | 0.01 | 1.9 | 0.01 | 2.3 | 0.02 | 7.6 | 0.03 | 10.0 | | Positive Control | 2.01 | 0.05 | 2.7 | 0.03 | 1.6 | 0.02 | 1.1 | 0.00 | 0.2 | 0.02 | 0.8 | 0.07 | 3.4 | ${}^{a}$ Standard deviation of mean concentration (Index) b Coefficient of variation PMA P200017: FDA Summary of Safety and Effectiveness Data {11} The instrument family members XP, XPT and CPT demonstrated equivalent reproducibility (Table 4). The instruments demonstrated only minor variability in the components evaluated. Table 4. ADVIA Centaur XP, XPT and CP - Summary of ADVIA Centaur Anti-HBe2 Reproducibility Results | System | Sample | % CV Reproducibility | | --- | --- | --- | | ADVIA CENTAUR XP | Serum 1 | 8.3% | | | Serum 2 | 6.7% | | | Serum 3 | 2.3% | | | Serum 4 | 3.2% | | | Negative Control | 8.2% | | | Positive Control | 2.3% | | ADVIA CENTAUR XPT | Serum 1 | 8.8% | | | Serum 2 | 6.4% | | | Serum 3 | 2.5% | | | Serum 4 | 3.1% | | | Negative Control | 5.7% | | | Positive Control | 2.7% | | ADVIA CENTAUR CP | Serum 1 | 9.7% | | | Serum 2 | 6.8% | | | Serum 3 | 3.3% | | | Serum 4 | 3.8% | | | Negative Control | 10.0% | | | Positive Control | 3.4% | # Sensitivity/Seroconversion Panels The seroconversion sensitivity of the ADVIA Centaur Anti-HBe2 assay was evaluated by testing three commercially available HBV patient seroconversion panels in replicates on ADVIA Centaur XP, XPT, and CP instruments, and the FDA-approved reference assay. The performance of the PMA P200017: FDA Summary of Safety and Effectiveness Data {12} ADVIA Centaur Anti-HBe2 assay on all three instruments matched the performance of the reference assay (Table 5): Table 5. Summary of ADVIA Centaur Anti-HBe2 Panel Results | Panel ID | aHBe Reactive Result from Initial Draw Date | | ADVIA Centaur Assay vs. Reference Assay | | --- | --- | --- | --- | | | Reference Assay (Days) | ADVIA Centaur Assay (Days) | Difference in Bleed Days (Numbers)a | | 6510 | 70 | 70 | 0 | | SCP-HBV-001 | 88 | 88 | 0 | | SCP-HBV-002 | 74 | 74 | 0 | a 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 reactive. ## Analytical Sensitivity/Dilution Study with Standard The analytical sensitivity was assessed with two lots of ADVIA Centaur Anti-HBe2 kits by testing a series of dilutions of a Paul Ehrlich Institute (PEI) Standard (WHO 1st International Standard Anti-Hepatitis B virus e antigen) on ADVIA Centaur XP, XPT and CP systems. A stock solution of standard PEI 120 IU/mL was used to prepare dilution series using negative human serum at concentrations ranging from 1.00 to 0.00 PEI IU/mL. Linear regression was used to determine the concentration of the PEI reference sample that corresponds to the ADVIA Centaur Anti-HBe2 assay cut-off (Index = 1.00). Analytical sensitivity evaluated with two different reagent lots of ADVIA Centaur Anti-HBe2 on ADVIA Centaur XP, CP and XPT instruments only minor differences from the WHO International Standard in all cases, as shown below in Table 6. Table 6. ADVIA Centaur XP, XPT and CP - Concentration at the Cut-Off | Reagent Lot | IU/mL at Cut-Off | | | | --- | --- | --- | --- | | | Platform | | | | | ADVIA Centaur | ADVIA Centaur XPT | ADVIA Centaur CP | | B30919 | 0.17 | 0.17 | 0.16 | | B30775 | 0.17 | 0.18 | 0.16 | ## Endogenous Interference Testing was performed in accordance with CLSI Documents EP07-A3, Interference Testing in Clinical Chemistry, Third Edition, and EP37, Supplemental Tables for Interference Testing. PMA P200017: FDA Summary of Safety and Effectiveness Data Page 13 of 39 {13} The sensitivity of the ADVIA Centaur Anti-HBe2 assay to interference by endogenous substances was evaluated at the concentrations indicated in Table 7. The study was performed using one lot of reagents to test each interferant on an ADVIA Centaur XP system with serum, EDTA plasma and Lithium Heparin plasma matrices. The reactivity of a pool of samples was spiked with the substance being tested (Test) and compared to the reactivity of the same pool of samples spiked with the solvent or media (Control) used for the substance. The bias was calculated as percentage of (Test-Control)/Control. The results of the interference study concluded that the following substances at the tested concentrations do not interfere with the ADVIA Centaur Anti-HBe2 assay using the three tested sample matrices (serum, EDTA plasma and Lithium Heparin plasma). Table 7. Summary of Endogenous Substances used in the Interference Study | Substance | Substance Test Concentration Units | | --- | --- | | Bilirubin conjugated (icteric) | 40 mg/dL | | Bilirubin unconjugated (icteric) | 40 mg/dL | | Biotin | 3500 ng/mL | | Cholesterol | 400 mg/dL | | Hemoglobin (hemolyzed) | 1000 mg/dL | | Lipemia (glyceryl trioleate) | 3000 mg/dL | | Lipemia (intralipid) | 3000 mg/dL | | Protein (hyperproteinemic) | 12 g/dL | | Protein (hypoproteinemic) | 3 g/dL | ## Analytical Specificity (Cross-Reactivity) The ADVIA Centaur Anti-HBe2 assay was evaluated on ADVIA Centaur XP system for potential cross-reactivity with viral antibodies or other cross-reactants in the specimens from individuals with medical conditions unrelated to the HBV infection. A total of 314 samples from 33 unrelated medical conditions were tested in singlicate with one reagent lot of the ADVIA Centaur Anti-HBe2 assay and an FDA-approved Anti-HBe Assay (Reference Assay) to determine the true status of the samples. Three samples were identified as equivocal by ADVIA Centaur Anti-HBe2 in the first analysis and they were re-tested in duplicate. The repeated results from these three samples were concordant with reference Anti-HBe results. One HCV sample was identified as discrepant in the first analysis. The sample was analyzed in duplicate by ADVIA Centaur Anti-HBe2 and the reference assay and was confirmed as a discrepant sample. Finally, it was tested in singlicate by a PMA P200017: FDA Summary of Safety and Effectiveness Data Page 14 of 39 {14} second FDA-approved assay. The sample was resolved as concordant with ADVIA Centaur Anti-HBe2 assay. The Cross Reactivity by other disease conditions study demonstrates agreement between the ADVIA Centaur Anti-HBe2 assay and the Reference Assays (Table 8) except for the single HCV sample that was concordant with the second reference assay. Hence, no interference was observed with the concentrations tested. Table 8. Summary of Cross-Reactivity Testing | Clinical Category | Number Tested | Reactive | | | --- | --- | --- | --- | | | | ADVIA Centaur Assay | FDA-approved aHBe Assay | | Alanine amino transferase (ALT) | 10 | 0 | 0 | | Anti-nuclear antibodies (ANA) | 10 | 0 | 0 | | Cytomegalovirus (CMV) IgG | 6 | 0 | 0 | | Cytomegalovirus (CMV) IgM | 14 | 0 | 0 | | Dialysis patients | 10 | 0 | 0 | | Escherichia coli | 10 | 0 | 0 | | Epstein Barr Virus (EBV) VCA IgG | 10 | 0 | 0 | | Epstein Barr Virus (EBV) VCA IgM | 10 | 0 | 0 | | Flu vaccinated | 10 | 0 | 0 | | HAV IgG Total | 10 | 1 | 1 | | HBsAg vaccinated | 10 | 1 | 1 | | Hepatitis C Virus (HCV) | 10 | 1 | 1* | | Herpes Simplex Virus (HSV-1) | 10 | 0 | 0 | | Herpes Simplex Virus (HSV-2) | 10 | 0 | 0 | | Heterophile antibodies | 10 | 0 | 0 | | Human Herpes Virus 6 (HHV-6) | 9 | 0 | 0 | | Human Herpes Virus 8 (HHV-8) | 10 | 0 | 0 | | Human Immunodeficiency Virus -1 | 10 | 0 | 0 | | Human Immunodeficiency Virus -2 | 10 | 3 | 3 | | Human Anti-Mouse Antibodies (HAMA) | 10 | 0 | 0 | | Multi-parous females | 10 | 1 | 1 | | Multiple Myeloma | 10 | 0 | 0 | | Non-viral liver disease | 10 | 0 | 0 | | Pregnant females | 10 | 0 | 0 | | Rheumatoid Arthritis | 10 | 0 | 0 | | Rheumatoid Factor (RF) | 10 | 1 | 1 | | Rubella IgG | 10 | 0 | 0 | PMA P200017: FDA Summary of Safety and Effectiveness Data Page 15 of 39 {15} | Syphilis (total or IgG) | 10 | 1 | 1 | | --- | --- | --- | --- | | Toxoplasma gondii IgG | 10 | 0 | 0 | | Toxoplasma gondii IgM | 5 | 0 | 0 | | Urinary Tract Infection (UTI) | 10 | 1 | 1 | | Varicella zoster virus (VZV) IgG | 5 | 0 | 0 | | Varicella zoster virus (VZV) IgM | 5 | 0 | 0 | * concordant with the second reference assay # Sample Equivalence/Matrix Effect The study was performed on the ADVIA Centaur XP instrument to assess the influence of different matrices on the results of the ADVIA Centaur Anti-HBe2 assay. Fifty matched sets of samples of different matrices (serum, serum separator tube (SST), EDTA plasma and Lithium Heparin plasma) from commercial sources were analyzed in duplicate in randomized order. The titers of the 25 reactive (&gt;1 Index) and 25 non-reactive (&lt;1 Index) samples were distributed throughout the range of the assay with 30 spanning the cutoff (0.70-1.60 Index). Because of the difficulty in obtaining samples throughout the ADVIA Centaur Anti-HBe2 assay measuring range (0.00-3.50 Index), 46 sets were prepared by spiking negative samples with different quantities of positive samples. The quantity of spiking material was always $&lt; 10\%$ of the sample volume. The results for all matrices were plotted on a XY graph compared to the values obtained from serum samples and weighted Deming regression fit was used to evaluate the variability. The correlation coefficient was calculated using Pearson correlation. Matrix equivalency was determined in accordance with CLSI Document EP09-A3, Measurement Procedure Comparison and Bias Estimation Using Patient Samples; Approved Guideline—Third Edition. All samples demonstrated acceptable differences between matrices throughout the measurement range (Table 9). Table 9. Summary of ADVIA Centaur Anti-HBe2 Sample Matrix Results | Tube | N | Sample Interval | Slope | Intercept (Index) | r | | --- | --- | --- | --- | --- | --- | | Dipotassium EDTA (plasma) | 50 | 0.09–2.04 Index | 0.93 | -0.01 | 0.99 | | Lithium heparin (plasma) | 50 | 0.08–2.08 Index | 0.91 | -0.01 | 0.99 | | Gel-barrier tube (serum) | 50 | 0.13–2.24 Index | 1.02 | 0.01 | 0.99 | # Reagent Carry-Over Study To evaluate the specific susceptibility of the Anti-HBe2 and HBc assay carry-over, a study was performed to identify potential interferences due to reagent carryover between assays that are run- PMA P200017: FDA Summary of Safety and Effectiveness Data {16} in random-access mode, and to define mitigations required to permit assays that share the same reagent/ancillary probe(s) to be run-in random-access mode. The reagent packs of ADVIA Centaur Anti-HBe2 were spiked into the assay reagents for different assays that use the same probe and conversely, the components of different ADVIA Centaur XP/CP assays were spiked into the new ADVIA Centaur Anti-HBe2 reagent packs for testing. Controls (Positive and Negative) were used as the samples in the study. Results indicated that the Anti-HBe2 assay affected the HBeAg, TSTO (Testosterone) and HBcT assays. Mitigation using a Wash Pack in addition to the default water wash were successfully validated for all interferences observed and were implemented into the ADVIA Centaur XP and ADVIA Centaur CP Test Definitions. After mitigation no carryover was detected. ## Stability Studies ### a. Sample Stability Studies were performed to determine the stability of patient samples under different storage conditions. Three matrices (serum or serum separator tube (SST), EDTA plasma, and Lithium-Heparin plasma) were analyzed on an ADVIA Centaur XP instrument with one lot of ADVIA Centaur Anti-HBe2 assay reagents. The effect of multiple freeze/thaw cycles and time to centrifugation on the stability of the samples was also evaluated. Ten samples per matrix per study were used to evaluate sample handling and storage temperature, and freeze/thaw cycles. Fifteen samples/matrix were used for time to centrifugation studies, with the following distribution: - 3 Non-reactive samples (≤ 0.69 Index) - 2 samples between 0.70–1.00 Index (Below Cut off) - 3 samples between 1.00–1.60 Index (Low Reactive) - 2 samples &gt; 1.60 Index. The results of the sample handling and storage temperature studies, freeze/thaw, and long-term deep frozen (-70°C &amp; -20°C) studies supported the claims for stability for all samples and matrices (serum or SST, EDTA plasma and Lithium Heparin plasma) (Table 10). Table 10. Summary of the Results for the Sample Stability Study Types | Stability Study Type | | Expiry | | --- | --- | --- | | | Room Temperature (Nominal 25°C) | 72 hours | PMA P200017: FDA Summary of Safety and Effectiveness Data Page 17 of 39 {17} | Handling and Storage Temperature Studies | Refrigerated (2–8°C) in secondary containers | 8 days | | --- | --- | --- | | | Frozen (Nominal -20°C) | 12 months | | | Deep Frozen (Nominal -70°C) | 12 months | | | On Board Sample Stability | 4 hours | | | Primary Tube (“On the Clot”) | 7 | | Freeze/Thaw Cycles | Freeze Thaw (-20°C) | 3 cycles | | | Freeze Thaw (-70°C) | 3 cycles | | Time to Centrifugation | | 24 hours | ## B. Ambient Temperature Sensitivity The accuracy of the test was evaluated on ADVIA Centaur XP and CP platforms to determine the effect of the ambient temperature changes (laboratory temperature) on assay performance. Ambient Temperature Sensitivity (ATS) was assessed at three different temperature set points: 18°C, 24°C and 30°C. All results demonstrated equivalent performance at ambient temperatures between 18°C and 30°C. ## C. Reagent Stability ### Reagent Real Time Stability Study (Shelf-Life) Real time stability testing was performed on three lots of ADVIA Centaur Anti-HBe2 (reagent ReadyPack and kitted Calibrators) on ADVIA Centaur XP instrument to ascertain its shelf-life. Each lot was stored at 2–8°C and was tested at pre-determined checkpoints from T = 0 to 13 months. This is an on-going study. The results support the current shelf-life claim of 12 months for ADVIA Centaur Anti-HBe2 assay at 2–8°C. ### Stability of the Calibration Interval The stability of the working calibration curve was evaluated on the ADVIA Centaur XP and CP instruments with two lots of ADVIA Centaur Anti-HBe2. On day 0, eight ReadyPacks of the ADVIA Centaur Anti-HBe2 were placed on-board the ADVIA Centaur instrument and pierced. These ReadyPacks were kept on the instrument throughout the duration of the study. Moreover, on day 0 and every other checkpoint up to 61 days, one fresh ReadyPack was also loaded on the instrument. On each checkpoint, the Calibrators, Controls and Sample levels were randomly tested in triplicate using the on-board and the fresh ReadyPacks. The stability of the calibration was evaluated up to day 61. The results support the calibration interval claim of 21 days on ADVIA Centaur XP and CP instruments. PMA P200017: FDA Summary of Safety and Effectiveness Data Page 18 of 39 {18} PMA P200017: FDA Summary of Safety and Effectiveness Data Page 19 of 39 ## Reagent On-Board Stability Studies were performed with two lots of ADVIA Centaur Anti-HBe2 to establish the maximum time that an opened ReadyPack can be stored on-board the ADVIA Centaur XP and CP instruments. On day 0, eight ReadyPacks of ADVIA Centaur Anti-HBe2, considered as opened packs, were placed on-board the instrument and pierced. The ReadyPacks were kept on the instrument throughout the duration of the study. Moreover, on day 0 and every other check day up to 61 days, one fresh ReadyPack was also tested on the instrument. Analysis of the Calibrators, Controls and Sample levels were performed with both the on-board and fresh packs in triplicate on day 0 and every checkpoint up to 61 days. The results support the Reagent on-board stability claim of 60 days on ADVIA Centaur XP and CP instruments. ## Calibrators Open Vial Stability The study was performed with two lots of the ADVIA Centaur Anti-HBe2 on ADVIA Centaur XP instrument to establish the open vial stability at 2–8°C of Calibrators versus the unopened vials. One calibration with each Opened Vial Calibrators set (OV: opened and squeezed each check day and then kept at 2–8°C) and one calibration with the Unopened Vial Calibrators set (UV: also kept at 2–8°C) were performed. An immutable (kept at below -70°C) set of controls with each calibration was analyzed in triplicate on day 0 and every other check day up to 61 days. The results support the Open Vial stability claim of 60 days for ADVIA Centaur Anti-HBe2 Calibrators. ## Calibrator On-Board Stability The on-board stability of Calibrators was evaluated with two lots of the Calibrators on one ADVIA Centaur XP instrument. The results of the controls were within the acceptance range on the calibration obtained up to 9 hours. When Calibrators were run as samples, all results met the acceptance criteria up to 9 hours. The results support the on-board stability claim of 8 hours for the ADVIA Centaur Anti-HBe2 Calibrators on the ADVIA Centaur XP and CP instruments. ## Animal Studies Not applicable. ## Additional studies Not applicable. ## X. SUMMARY OF PRIMARY CLINICAL STUDIES The applicant performed a clinical study to establish a reasonable assurance of safety and effectiveness of the ADVIA Centaur anti-HBe2. Data from this clinical study were the basis for the PMA approval decision. A summary of the clinical study is presented below. {19} PMA P200017: FDA Summary of Safety and Effectiveness Data Page 20 of 39 ## A. Study Design A multisite study was conducted to evaluate the performance of the ADVIA Centaur anti-HBe2 assay. The study consists of a Qualitative Method Comparison: Agreement with a reference assay (FDA-approved Anti-HBe assay). The testing of the ADVIA Centaur Anti-HBe2 assay on the ADVIA Centaur XP, XPT and CP instruments, and the HBV serological classification were conducted at three external US sites with three reagent lots. The subject enrollment/sample collection was performed at or coordinated by four US clinical sites. ## B. Study Population Demographics and Baseline Parameters A total of 1877 subjects were enrolled for this study and prospective serum samples were collected. These included 180 pediatric subjects, the samples of 63 of which were spiked with adult reactive anti-HBe serum to generate contrived samples. Therefore, the prospective study population consisted of 1814 samples from unique patients including pediatric subjects (2 – 21 years old) as well as pregnant women (228). The samples from the pediatric subjects (&lt;11–20, Table 16) were tested on the XP instrument only; therefore, there were 1697 subjects tested with the CP instrument. The study population was considered either at high-risk for hepatitis B (due to lifestyle, behavior, occupation, or known exposure events) or was considered to have signs and symptoms of infection. The patients were divided by sex (55.5% female, 44.5% male). The study population was 27.1% Caucasian, 68.5% Black, 2.0% Asian, and 2.4% from unknown or other race. The mean age was 44 years (3–92 years). Patients in the study population were from the following geographic regions: Florida (34%), California (23.9%), Maryland (22.1%), Tennessee (6.4%), Georgia (5.3%), Nevada (3.9%), Massachusetts (3.5%), and other locations (0.9%). ADVIA Centaur Anti-HBe2 assay was tested on all three platforms, ADVIA Centaur XP, XPT and CP systems and the agreement calculated against an FDA-approved aHBe reference assay. An overview of the detailed results in the following tables of the ADVIA Centaur Anti-HBe2 Percent Positive Agreement (PPA) and Negative Percentage of Agreement (NPA) versus the reference assay on the different instrument systems is below (Table 11). ### Percent Agreement: Positive and negative percent agreement between the ADVIA Centaur Anti-HBe2 assay and the FDA approved comparator anti-HBe test were calculated for subjects by HBV disease classification. Positive percent agreement with the comparator anti-HBe test was determined by dividing the number of reactive ADVIA Centaur Anti-HBe2 results by the total number of subjects positive with the comparator anti-HBe test. Negative percent agreement with the comparator anti-HBe test was determined by dividing the number of negative ADVIA Centaur Anti-HBe2 results by the total number of subjects negative with the comparator anti-HBe test. {20} Table 11. ADVIA Centaur XP, XPT and CP - ADVIA Centaur Anti-HBe2 Clinical Performance Results including Unknown Serostatus Samples | System | Parameter | Study Result | 95% CI | | --- | --- | --- | --- | | ADVIA Centaur XP | PPA | 97.6% (163/176) | 87.5%-96.2% | | | NPA | 99.0% (1621/1638) | 98.4%-99.4% | | ADVIA Centaur XPT | PPA | 92.0% (162/176) | 87.1%-95.2% | | | NPA | 99.1% (1507/1521) | 98.5%-99.5% | | ADVIA Centaur CP | PPA | 93.2% (164/176) | 88.5%-96.1% | | | NPA | 99.0% (1506/1521) | 98.4%-99.4% | # HBV Disease Classification Patients were assessed for hepatitis markers using commercially available, FDA-approved reference assays. Each patient's HBV infection status was determined based on a single specimen and the reactive $(+)$ / nonreactive $(-)$ patterns of six HBV reference serological markers obtained from two tests (Table 15): hepatitis B virus surface antigen (HBsAg), hepatitis B virus e antigen (HBeAg), total antibody to hepatitis B virus core antigen (anti-HBc Total), IgM antibody to hepatitis B core antigen (anti-HBc IgM), total antibody to HBeAg (anti-HBe), and total antibody to hepatitis B virus surface antigen (anti-HBs). There were 26 unique reference marker patterns observed using the FDA-approved assays (Table 12). Table 12. HBV Classification by Reference Markers (All Testing Sites) including Unknown Serostatus Samples | HBV Classification | HBsAg | HBeAg | Anti-HBc IgM | Anti-HBc Total | Anti-HBs | FDA-Approved anti-HBe Assay | | --- | --- | --- | --- | --- | --- | --- | | Acute | +a | + | + | + | -b | - | | Chronic | + | + | - | + | + | - | | Chronic | + | + | - | + | - | - | | Chronic | + | - | - | + | - | + | | Chronic | + | - | - | + | - | - | | Early Recovery | - | - | Equivocal | + | + | + | | Early Recovery | - | - | + | + | + | + | PMA P200017: FDA Summary of Safety and Effectiveness Data {21} | Early Recovery | - | - | + | + | - | + | | --- | --- | --- | --- | --- | --- | --- | | Recovery | - | - | - | + | - | + | | Immune Natural Infection | - | - | - | + | + | + | | Recovered | - | - | - | + | + | - | | Recovered | - | - | - | + | - | - | | HBV Vaccine Response | - | - | - | - | + | - | | Not previously infected | - | - | - | - | - | - | | Unknown | + | + | - | - | - | - | | Unknown | + | + | Equivocal | + | - | - | | Unknown | - | + | - | - | + | - | | Unknown | - | + | - | - | - | - | | Unknown | - | + | - | + | + | - | | Unknown | - | + | - | + | - | - | | Unknown | + | - | - | - | + | - | | Unknown | - | - | - | - | - | + | | Unknown | - | - | Equivocal | + | - | - | | Unknown | - | - | + | - | - | - | | Unknown | - | - | - | - | + | + | | Unknown | - | - | + | + | + | - | a $+ =$ Reactive. b $-\equiv$ Nonreactive Samples from patients who fell into all of the categories: acute, chronic, early recovery, recovery, immune natural infection (recovery), recovered, HBV vaccine response, not previously infected, and unknown serostatus categories, were included in the study and used for data analysis. Thirty-eight samples returned a result of unknown serological status (2.09%; 38/1814). This is a possible outcome from patient samples evaluated for disease classification that may be encountered in the clinic, but it was not clear how these should be included in the analysis. The performance of the device was evaluated primarily with the samples included, with additional information on performance with the samples excluded in the footnotes below the tables. Expected Values PMA P200017: FDA Summary of Safety and Effectiveness Data {22} All of the samples from the 1814 unique patients including pediatric and pregnant women subjects were tested with the ADVIA Centaur Anti-HBe2. - 784 patients (43.2%) were from the population considered at risk for hepatitis B (high risk) due to lifestyle, behavior, occupation, or known exposure events. - 1030 patients (56.8%) were from the signs and symptoms population The ADVIA Centaur Anti-HBe2 results for the prospective population for all sites combined by age group and gender are summarized below including with and without the unknown serostatus samples (Table 13). Table 13. ADVIA Centaur XP - Distribution of Study Population by Age Group and Gender (All Testing Sites) including Unknown Serostatus Samples | Age Range (Years)* | Gender | Reactive | | Nonreactive | | Total Number Tested | Number without Unknown Serostatus Samples | | --- | --- | --- | --- | --- | --- | --- | --- | | | | N | % | N | % | | | | < 11 | Male | 0 | 0.0 | 11 | 100 | 11 | 11 | | | Female | 0 | 0.0 | 10 | 100 | 10 | 10 | | | Overall | 0 | 0.0 | 21 | 100 | 21 | 21 | | 11–20 | Male | 0 | 0.0 | 33 | 100 | 33 | 33 | | | Female | 1 | 1.7 | 58 | 98.3 | 59 | 58 | | | Overall | 1 | 1.1 | 91 | 98.9 | 92 | 91 | | 21–30 | Male | 0 | 0.0 | 97 | 100 | 97 | 96 | | | Female | 3 | 1.1 | 265 | 98.9 | 268 | 261 | | | Overall | 3 | 0.8 | 362 | 99.2 | 365 | 357 | | 31–40 | Male | 4 | 4.2 | 91 | 95.8 | 95 | 93 | | | Female | 8 | 4.1 | 188 | 95.9 | 196 | 191 | | | Overall | 12 | 4.1 | 279 | 95.9 | 291 | 284 | | 41–50 | Male | 17 | 11.6 | 129 | 88.4 | 146 | 143 | | | Female | 21 | 13.2 | 138 | 86.8 | 159 | 155 | | | Overall | 38 | 12.5 | 267 | 87.5 | 305 | 298 | | 51–60 | Male | 50 | 17.9 | 229 | 82.1 | 279 | 274 | | | Female | 37 | 17.8 | 171 | 82.2 | 208 | 206 | | | Overall | 87 | 17.9 | 400 | 82.1 | 487 | 480 | PMA P200017: FDA Summary of Safety and Effectiveness Data Page 23 of 39 {23} | Age Range (Years)* | Gender | Reactive | | Nonreactive | | Total Number Tested | Number without Unknown Serostatus Samples | | --- | --- | --- | --- | --- | --- | --- | --- | | | | N | % | N | % | | | | 61–70 | Male | 20 | 15.9 | 106 | 84.1 | 126 | 123 | | | Female | 14 | 15.6 | 76 | 84.4 | 90 | 88 | | | Overall | 34 | 15.7 | 182 | 84.3 | 216 | 211 | | 71–92 | Male | 3 | 14.3 | 18 | 85.7 | 21 | 21 | | | Female | 2 | 12.5 | 14 | 87.5 | 16 | 13 | | | Overall | 5 | 13.5 | 32 | 86.5 | 37 | 34 | | Total | Male | 94 | 11.6 | 714 | 88.4 | 808 | 794 | | | Female | 86 | 8.5 | 920 | 91.5 | 1006 | 982 | | | Overall | 180 | 9.9 | 1634 | 90.1 | 1814 | 1776 | *CP did not test samples from pediatric subjects ## C. Safety and Effectiveness Results ### Safety Results The safety of this device is related to the efficacy described below as incorrect results may lead to patient mismanagement. ### Efficacy Results #### Prospective Population The performance of the ADVIA Centaur Anti-HBe2 assay was evaluated against the risk groups, the disease classification, and the subpopulations. The performance on the XP and CP instruments is presented separately. The results from all testing sites are combined because the performance was equivalent across sites. #### a. By Risk Groups The performance of the ADVIA Centaur Anti-HBe2 assay was evaluated in the sign and symptom (Tables 14A, 14B and 14C) and high-risk (Tables 15A, 15B and 15C) prospective populations on each assay system and compared to the reference assay. The percent agreement and confidence intervals for the prospective population by risk group is presented below (Table 16A, 16B and 16C). PMA P200017: FDA Summary of Safety and Effectiveness Data Page 24 of 39 {24} 1) Signs and Symptom prospective population: Table 14A. ADVIA Centaur XP - Comparison of Results in the Signs and Symptoms Prospective Population including Unknown Serostatus Samples | Reference Assay | ADVIA Centaur Anti-HBe2 Assay | | | | --- | --- | --- | --- | | | Reactive | Nonreactive | Total | | Reactive | 111 | 8 | 119 | | Nonreactive | 11 | 900 | 911 | | Total | 122 | 908 | 1030 | Table 14B. ADVIA Centaur XPT - Comparison of Results in the Signs and Symptoms Prospective Population including Unknown Serostatus Samples | Reference Assay | ADVIA Centaur Anti-HBe2 Assay | | | | --- | --- | --- | --- | | | Reactive | Nonreactive | Total | | Reactive | 111 | 8 | 119 | | Nonreactive | 10 | 843 | 853 | | Total | 121 | 851 | 972 | Table 14C. ADVIA Centaur CP - Comparison of Results in the Signs and Symptoms Prospective Population including Unknown Serostatus Samples | Reference Assay | ADVIA Centaur Anti-HBe2 Assay | | | | --- | --- | --- | --- | | | Reactive | Nonreactive | Total | | Reactive | 111 | 8 | 119 | | Nonreactive | 9 | 844 | 853 | | Total | 120 | 852 | 972 | 2) High-Risk Prospective Population: PMA P200017: FDA Summary of Safety and Effectiveness Data Page 25 of 39 {25} Table 15A. ADVIA Centaur XP - Comparison of Results in the High-risk Prospective Population including Unknown Serostatus Samples | Reference Assay | ADVIA Centaur Anti-HBe2 Assay | | | | --- | --- | --- | --- | | | Reactive | Nonreactive | Total | | Reactive | 52 | 5 | 57 | | Nonreactive | 6 | 721 | 727 | | Total | 58 | 726 | 784 | Table 15B. ADVIA Centaur XPT - Comparison of Results in the High-risk Prospective Population including Unknown Serostatus Samples | Reference Assay | ADVIA Centaur Anti-HBe2 Assay | | | | --- | --- | --- | --- | | | Reactive | Nonreactive | Total | | Reactive | 51 | 6 | 57 | | Nonreactive | 4 | 664 | 668 | | Total | 55 | 670 | 725 | Table 15C. ADVIA Centaur CP - Comparison of Results in the High-risk Prospective Population including Unknown Serostatus Samples | Reference Assay | ADVIA Centaur Anti-HBe2 Assay | | | | --- | --- | --- | --- | | | Reactive | Nonreactive | Total | | Reactive | 53 | 4 | 57 | | Nonreactive | 6 | 662 | 668 | | Total | 59 | 666 | 725 | Table 16A. ADVIA Centaur XP - Percent Agreement and Confidence Intervals (all testing sites) for Risk Groups including Unknown Serostatus Samples | Groups | Positive Agreement | | Negative Agreement | | | --- | --- | --- | --- | --- | | | % (x/n) | 95% CI | % (x/n) | 95% CI | PMA P200017: FDA Summary of Safety and Effectiveness Data {26} | Signs and Symptoms | 93.3% (111/119) | 87.3%–96.6% | 98.8% (900/911) | 97.9%–99.3% | | --- | --- | --- | --- | --- | | High Risk | 91.2% (52/57) | 81.1%–96.2% | 99.2% (721/727) | 98.2%–99.6% | | Total | 92.6% (163/176) | 87.8%–95.6% | 99.0% (1621/1638) | 98.4%–99.4% | Table 16B. ADVIA Centaur XPT - Percent Agreement and Confidence Intervals (all testing sites) for Risk Groups including Unknown Serostatus Samples | Groups | Positive Agreement | | Negative Agreement | | | --- | --- | --- | --- | --- | | | % (x/n) | 95% CI | % (x/n) | 95% CI | | Signs and Symptoms | 93.3% (111/119) | 87.3%–96.6% | 98.8% (843/853) | 97.9%–99.4% | | High Risk | 89.5% (51/57) | 78.9%–95.1% | 99.4% (664/668) | 98.5%–99.8% | | Total | 92.0% (162/176) | 87.1%–95.2% | 99.1% (1507/1521) | 98.5%–99.5% | Table 16C. ADVIA Centaur CP - Percent Agreement and Confidence Intervals (all testing sites) for Risk Groups including Unknown Serostatus Samples | Groups | Positive Agreement | | Negative Agreement | | | --- | --- | --- | --- | --- | | | % (x/n) | 95% CI | % (x/n) | 95% CI | | Signs and Symptoms | 93.3% (111/119) | 87.3%–96.6% | 98.9% (844/853) | 98.0%–99.4% | | High Risk | 93.0% (53/57) | 83.3%–97.2% | 99.1% (662/668) | 98.1%–99.6% | | Total | 93.2% (164/176) | 88.5%–96.1% | 99.0% (1506/1521) | 98.4%–99.4% | For the signs and symptoms population: excluding the unknown serostatus samples, the total PPA is 98.2 (111/113) (95%CI 93.8%–99.5%) and total NPA is 98.8% (883/894) (95% CI 97.8%–99.3%). For the high-risk population: excluding the unknown serostatus samples for the high-risk prospective population, the total PPA is 96.3% (52/54) (95%CI 87.5%–99.0%) and total NPA is 99.3% (710/715) (95% CI 98.4%–99.7%). PMA P200017: FDA Summary of Safety and Effectiveness Data Page 27 of 39 {27} All instrument systems demonstrated acceptable performance when compared with the reference assay. As expected, the agreement increased when the results were analyzed by the excluding 38 samples that did not fit into pre-determined serological status categories based on the patterns of the six HBV reference serological markers. ## b. By HBV Serological Classification ### 1) Results on ADVIA Centaur XP A total of 1814 samples including the 38 unknown serostatus samples were evaluated using the ADVIA Centaur Anti-HBe2 assay on the ADVIA Centaur XP and a reference aHBe assay for each sample classification (Table 17). The agreement and 95% CIs between the ADVIA Centaur Anti-HBe2 assay and a reference aHBe assay for each HBV classification are presented in the Table 18. Table 17. ADVIA Centaur XP - Results of Prospective Population by HBV Serological Classification including Unknown Serostatus Samples | HBV classification | Reference Assay-Reactive | | Reference Assay-Nonreactive | | Total | | --- | --- | --- | --- | --- | --- | | | ADVIA Centaur Anti-HBe2 Assay | | ADVIA Centaur Anti-HBe2 Assay | | | | | Reactive | Nonreactive | Reactive | Nonreactive | | | | N | N | N | N | N | | Acute | 0 | 0 | 0 | 2 | 2 | | Chronic | 33 | 0 | 2 | 23 | 58 | | Early Recovery | 9 | 0 | 0 | 0 | 9 | | Recovery | 15 | 0 | 0 | 0 | 15 | | Immune Natural Infection (Recovery) | 106 | 4 | 0 | 0 | 110 | | Recovered | 0 | 0 | 14 | 227 | 241 | | HBV Vaccine Response | 0 | 0 | 0 | 597 | 597 | | Not previously infected | 0 | 0 | 0 | 744 | 744 | | Unknown | 0 | 9 | 1 | 28 | 38 | | Total | 163 | 13 | 17 | 1621 | 1814 | PMA P200017: FDA Summary of Safety and Effectiveness Data Page 28 of 39 {28} Table 18. ADVIA Centaur XP - Percent Agreement and Confidence Intervals in the Prospective Population by HBV Serological Classification including Unknown Serostatus samples | HBV classification | Positive Agreement | | Negative Agreement | | | --- | --- | --- | --- | --- | | | % (x/n) | 95% CI | % (x/n) | 95% CI | | Acute | – | – | 100% (2/2) | 34.2%–100% | | Chronic | 100% (33/33) | 89.6%–100% | 92.0% (23/25) | 75.0%–97.8% | | Early Recovery | 100% (9/9) | 70.1%–100% | – | – | | Recovery | 100% (15/15) | 79.6%–100% | – | – | | Immune natural infection (Recovery) | 96.4% (106/110) | 91.0%–98.6% | – | – | | Recovered | – | – | 94.2% (227/241) | 90.5%–96.5% | | HBV Vaccine Response | – | – | 100% (597/597) | 99.4%–100% | | Not previously infected | – | – | 100% (744/744) | 99.5%–100% | | Unknown | 0.0% (0/9) | 0.0%–29.9% | 96.6% (28/29) | 82.8%–99.4% | | Total | 92.6% (163/176) | 87.8%–95.6% | 99.0% (1621/1638) | 98.3%–99.4% | ## 2) Results on ADVIA Centaur XPT A total of 1697 samples were run using the ADVIA Centaur Anti-HBe2 assay on the ADVIA Centaur XPT and a reference aHBe assay for each HBV serological classification. Comparison of results is shown in Table 19 for all testing sites. The agreement and 95% CIs between the ADVIA Centaur Anti-HBe2 assay and a reference aHBe assay for each HBV classification are presented in the Table 20. Table 19. ADVIA Centaur XPT - Results of Prospective Population by HBV Serological Classification including Unknown Serostatus Samples | HBV classification | Reference Assay-Reactive | Reference Assay-Nonreactive | Total | | --- | --- | --- | --- | | | ADVIA Centaur Anti-HBe2 Assay | ADVIA Centaur Anti-HBe2 Assay | | PMA P200017: FDA Summary of Safety and Effectiveness Data {29} Table 20. ADVIA Centaur XPT- Percent Agreement and Confidence Intervals in the Prospective Population by HBV Serological Classification including Unknown Serostatus samples | HBV classification | Positive Agreement | | Negative Agreement | | | --- | --- | --- | --- | --- | | | % (x/n) | 95% CI | % (x/n) | 95% CI | | Acute | - | - | 100% (2/2) | 34.2%-100% | | Chronic | 100% (33/33) | 89.6%-100% | 92.0% (23/25) | 75.0%-97.8% | | Early Recovery | 100% (9/9) | 70.1%-100% | - | - | | Recovery | 100% (15/15) | 79.6%-100% | - | - | | Immune natural infection (Recovery) | 95.5% (105/110) | 91.0%-98.6% | - | - | | Recovered | - | - | 95.4% (228/239) | 91.9%-97.4% | | HBV Vaccine Response | - | - | 100% (550/550) | 99.3%-100% | | Not previously infected | - | - | 100% (676/676) | 99.4%-100% | PMA P200017: FDA Summary of Safety and Effectiveness Data {30} | Unknown | 0.0% (0/9) | 0.0%-29.9% | 96.6% (28/29) | 82.8%-99.4% | | --- | --- | --- | --- | --- | | Total | 92.0% (162/176) | 87.1%-95.2% | 99.0% (1507/1521) | 98.5%-99.5% | # 3) Results on ADVIA Centaur CP A total of 1697 samples including pregnant women were run using the ADVIA Centaur Anti-HBe2 assay on the ADVIA Centaur CP and a reference aHBe assay for each HBV serological classification. A comparison of results in the prospective population by HBV serological classification is given below in the Table 21 with the unknown serostatus samples. The agreement and $95\%$ CIs between the ADVIA Centaur Anti-HBe2 assay and a reference aHBe assay for each HBV classification are presented in the Table 22. Table 21. ADVIA Centaur CP - Results of Prospective Population by HBV Serological Classification including Unknown Serostatus Samples | HBV classification | Reference Assay-Reactive | | Reference Assay-Nonreactive | | Total | | --- | --- | --- | --- | --- | --- | | | ADVIA Centaur Anti-HBe2 Assay | | ADVIA Centaur Anti-HBe2 Assay | | | | | Reactive | Nonreactive | Reactive | Nonreactive | | | | N | N | N | N | N | | Acute | 0 | 0 | 0 | 2 | 2 | | Chronic | 33 | 0 | 2 | 23 | 58 | | Early Recovery | 9 | 0 | 0 | 0 | 9 | | Recovery | 15 | 0 | 0 | 0 | 15 | | Immune Natural Infection (Recovery) | 107 | 3 | 0 | 0 | 110 | | Recovered | 0 | 0 | 12 | 227 | 239 | | HBV Vaccine Response | 0 | 0 | 0 | 550 | 550 | | Not previously infected | 0 | 0 | 0 | 676 | 676 | | Unknown | 0 | 9 | 1 | 28 | 38 | | Total | 164 | 12 | 15 | 1506 | 1697 | PMA P200017: FDA Summary of Safety and Effectiveness Data {31} Table 22. ADVIA Centaur CP - Percent Agreement and Confidence Intervals in the Prospective Population by HBV Serological Classification including Unknown Serostatus samples | HBV classification | Positive Agreement | | Negative Agreement | | | --- | --- | --- | --- | --- | | | % (x/n) | 95% CI | % (x/n) | 95% CI | | Acute | – | – | 100% (2/2) | 34.2%–100% | | Chronic | 100% (33/33) | 89.6%–100% | 92.0% (23/25) | 75.0%–97.8% | | Early Recovery | 100% (9/9) | 70.1%–100% | – | – | | Recovery | 100% (15/15) | 79.6%–100% | – | – | | Immune natural infection (Recovery) | 97.3% (107/110) | 92.3%–99.1% | – | – | | Recovered | – | – | 95.0% (227/239) | 91.4%–97.1% | | HBV Vaccine Response | – | – | 100% (550/550) | 99.3%–100% | | Not previously infected | – | – | 100% (676/676) | 99.4%–00% | | Unknown | 0.0% (0/9) | 0.0%–29.9% | 96.6% (28/29) | 82.8–99.4% | | Total | 93.2% (164/176) | 88.5%–96.1% | 99.0% (1506/1521) | 98.4%–99.4% | The data presented from all three systems demonstrated acceptable performance of the Anti-HBe2 assay when compared with an FDA-approved reference assay ## Pregnant Population Serum samples were tested from 228 women with either signs and symptoms of HBV infection or with risk factors for HBV infection, who were in the first (90/228, 39.5%), second (71/228, 31.1%), or third trimester (67/228, 29.4%) of pregnancy. The results were evaluated based on trimester (Table 23) and the PPA and NPA between the ADVIA Centaur Anti-HBe2 assay and the reference anti-HBe assay for each trimester is demonstrated in the Table 24. Table 23. ADVIA Centaur XP, XPT and CP - Summary of Pregnant Population including Unknown Serostatus Samples PMA P200017: FDA Summary of Safety and Effectiveness Data Page 32 of 39 {32} | Trimester | Reference Assay - Reactive | | Reference Assay - Nonreactive | | Total | | --- | --- | --- | --- | --- | --- | | | ADVIA Centaur Anti-HBe2 | | ADVIA Centaur Anti-HBe2 | | | | | Reactive | Nonreactive | Reactive | Nonreactive | | | | N | N | N | N | | | First | 0 | 0 | 0 | 90 | 90 | | Second | 0 | 0 | 0 | 71 | 71 | | Third | 2 | 1 | 0 | 64 | 67 | | Total | 2 | 1 | 0 | 225 | 228 | Table 24. ADVIA Centaur XP, XPT and CP - Percent Agreement and Confidence Intervals: Pregnant Population (all testing sites) including Unknown Serostatus Samples | Trimester | Positive Agreement | | Negative Agreement | | | --- | --- | --- | --- | --- | | | % (x/n) | 95% CI | % (x/n) | 95% CI | | First | - | - | 100% (90/90) | 95.9%–100% | | Second | - | - | 100% (71/71) | 94.9%–100% | | Third | 66.7% (2/3) | 20.8%–93.9% | 100% (64/64) | 94.3%–100% | | Total | 66.7% (2/3) | 20.8%–93.9% | 100% (225/225) | 98.3%–100% | These data indicate that the performance of the assay for detecting anti-HBe is acceptable in all trimesters of pregnancy on all three systems. ## Pediatric Population Pediatric (non-pregnant) samples were prospectively collected (N = 180). Due to the limited availability/occurrence of anti-HBe positive pediatric samples, the clinical study was supplemented with contrived samples to assess the positive percentage of agreement between ADVIA Centaur Anti-HBe2 and reference assay in pediatric population. The 63 contrived samples were prepared by spiking individual adult high positive samples into negative pediatric samples obtained from unique high-risk pediatric patients. The titer of the spiked samples ranged from 1.25 and &gt;3.50 Index. The pediatric population was only tested on the ADVIA Centaur XP system due to volume limitations on those samples. The pediatric population analysis was stratified by two age groups: 2–12 years and 13–21 years. An evaluation of the performance of the assay on pediatric population (Table 25) and agreement and the 95% CI between the ADVIA Centaur Anti-HBe2 assay and a reference aHBe assay are presented (Table 26). PMA P200017: FDA Summary of Safety and Effectiveness Data Page 33 of 39 {33} Table 25. ADVIA Centaur XP - Summary of Pediatric Populations (all testing sites) | Age Range (Years) | Reference Assay - Reactive | | Reference Assay - Nonreactive | | Total N | | --- | --- | --- | --- | --- | --- | | | ADVIA Centaur Anti-HBe2 Assay | | ADVIA Centaur Anti-HBe2 Assay | | | | | Reactive | Nonreactive | Reactive | Nonreactive | | | | N | N | N | N | | | 2-12 | 33 | 0 | 0 | 27 | 60 | | 13-21 | 30 | 0 | 1 | 89 | 120 | | Total | 63 | 0 | 1 | 116 | 180 | Table 26. ADVIA Centaur XP - Percent Agreement and Confidence Intervals: Pediatric Population (all testing sites) | Age Range (Years) | Positive Agreement | | Negative Agreement | | | --- | --- | --- | --- | --- | | | % (x/n) | 95% CI | % (x/n) | 95% CI | | 2-12 | 100% (33/33) | 89.6%–100% | 100% (27/27) | 87.5%–100% | | 13-21 | 100% (30/30) | 88.6%–100% | 98.9% (89/90) | 94.0%–99.8% | | Total | 100% (63/63) | 94.3%–100% | 99.1% (116/117) | 95.3%–99.8% | Comparison of the ADVIA Centaur XPT System and ADVIA Centaur XP System: Because the ADVIA Centaur XP and ADVIA Centaur XPT systems are family members, the percent agreement between the instrument family members ADVIA Centaur XPT system and the ADVIA Centaur XP system was evaluated by testing 1697 (with unknown serostatus) and 1659 (without unknown serostatus) samples at three clinical testing sites. Each site used one ADVIA Centaur XPT system and one ADVIA Centaur XP system, and tested three lots of reagents. The samples included patients with general signs and symptoms of hepatitis or a high risk of HBV infection, including pregnant women. The results are given below in the Table 27 (unknown serostatus included) and Table 28 (unknown serostatus excluded) Table 27. ADVIA Centaur XPT System versus ADVIA Centaur XP System with included Unknown Serostatus Samples | ADVIA Centaur XPT | ADVIA Centaur XP | | | | --- | --- | --- | --- | | | Reactive | Nonreactive | Total | PMA P200017: FDA Summary of Safety and Effectiveness Data {34} The PPA was 97.8% (175/179) (95% CI 94.4% to 99.1%). The NPA was 99.9% (1517/1518) (95%CI 99.6% to 100%). These data indicate that the performance of the ADVIA Centaur Anti-HBe2 assay is equivalent on the ADVIA Centaur XP and ADVIA Centaur XPT instruments. Table 28. ADVIA Centaur XPT System versus ADVIA Centaur XP System with excluded Unknown Serostatus samples | ADVIA Centaur XPT | ADVIA Centaur XP | | | | --- | --- | --- | --- | | | Reactive | Nonreactive | Total | | Reactive | 174 | 1 | 175 | | Nonreactive | 4 | 1480 | 1484 | | Total | 178 | 1481 | 1659 | The PPA was 97.8% (174/178) (95% CI 94.4% to 99.1%). The NPA was 99.9% (1480/1481) (95% CI 99.6% to 100%). These data indicate that the performance of the ADVIA Centaur Anti-HBe2 assay is equivalent on the ADVIA Centaur XP and ADVIA Centaur XPT instruments. ## D. Financial Disclosure The Financial Disclosure by Clinical Investigators regulation (21 CFR 54) requires applicants who submit a marketing application to include certain information concerning the compensation to, and financial interests and arrangement of, any clinical investigator conducting clinical studies covered by the regulation. The clinical study included three investigators. None of the clinical investigators were full-time or part-time employees of the sponsor and all three investigators had disclosable financial interests/arrangements as defined in sections 54.2(a), (b), (c), and (f). The information provided does not raise any questions about the reliability of the data. ## XI. PANEL MEETING RECOMMENDATION AND FDA'S POST-PANEL ACTION In accordance with the provision of section 515(c)(3) 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 duplicates information previously reviewed by this panel. PMA P200017: FDA Summary of Safety and Effectiveness Data Page 35 of 39 {35} PMA P200017: FDA Summary of Safety and Effectiveness Data Page 36 of 39 # XII. CONCLUSIONS DRAWN FROM PRECLINICAL AND CLINICAL STUDIES ## A. Safety Conclusions The risk of the device is based on data collected in the non-clinical and clinical studies conducted to support PMA approval as described above. Based on the results from both studies, ADVIA Centaur anti-HBe2 assay, when used according to the provided directions and in conjunction with all relevant clinical and laboratory findings, should be safe to use and poses minimal risk to the patient due to false test results. ## B. Effectiveness Conclusions The effectiveness of the ADVIA Centaur anti-HBe2 assay has been demonstrated by the sensitivity and specificity which has been comparable with the current commercially available FDA-approved anti-HBe assay among all populations tested. The results from both the non-clinical and clinical studies indicate that the ADVIA Centaur Anti-HBe2 assay is safe and effective for the in vitro qualitative detection of antibodies to the hepatitis B e antigen (Anti-HBe) in adults and pediatric serum and plasma (EDTA and Lithium Heparin). ## C. Benefit-Risk Determination The probable benefits of the device are also based on data collected on data collected in the clinical study conducted to support PMA approval as described above. The Hepatitis B Virus (HBV) represents a major global health concern. It causes a number of liver diseases, ranging from acute and chronic hepatitis to cirrhosis and primary liver cancer. Hepatitis B envelope (e) antigen (HBeAg) is found in the early phase of hepatitis B infection soon after hepatitis B surface antigen (HBsAg) becomes detectable. Serum levels of both antigens rise rapidly during the period of viral replication. The presence of HBeAg correlates with hepatitis B virus (HBV) infectivity, the number of viral Dane particles, the presence of core antigen in the nucleus of the hepatocyte, and presence of viral DNA polymerase in serum. During recovery from acute hepatitis B, after HBeAg level declines and becomes undetectable, HBe antibody (anti-HBe) appears in the serum. Anti-HBe usually remains detectable for several years after recovery from acute infection. In HBV carriers and chronic hepatitis B patients, positive HBeAg results usually indicate presence of active HBV replication and high infectivity. A negative HBeAg result indicates very minimal or lack of HBV replication. Positive anti-HBe results usually indicate inactivity of the virus and low infectivity. Positive anti-HBe results usually indicate inactivity of the virus and low infectivity. Positive anti-HBe results in the presence of detectable HBV DNA in serum indicate active viral replication in these patients. The benefit of the assay to detect antibodies to the HBeAg can be used to monitor the progress of hepatitis B viral infection. The assay will be used as an aid in the management of patients with chronic HBV infection undergoing antiviral therapy. Treatment with antiviral in timely manner may result improved morbidity and mortality in infected patients. The clinical studies suggest that patients will benefit from the assay. {36} The probable risks of the device are also based on the data collected in the clinical study conducted to support PMA approval as described above. The risks associated with the device, when used as intended, are those related to the risk of false test results, failure to correctly interpret the test results, and failure to correctly operate the device. Risks of erroneously high results include improper patient management, such as treatment for hepatitis B with antiviral medication. Administration of antiviral medication has risks including toxicity and more rarely allergic reactions. Risks of erroneously low results include improper patient management, such as potentially missing and under-treating a patient who has hepatitis B infection. Under-treating a patient with hepatitis B infection whose clinical picture warrants antiviral treatment could result in the known sequelae of HBV infection and may result in higher morbidity and mortality in these patients. Under-treating hepatitis B in patients whose clinical picture otherwise warrants treatment will lead to continued symptoms, increases in all-cause mortality, liver disease-related complications and death, hepato-cellular carcinoma rates, and need for liver transplantation. In conclusion, the clinical benefits outweigh the potential risks for the proposed assay, considering the mitigations of the risks provided in the premarket approval as well as general controls. The required premarket approval helps to ensure that errors will be uncommon and will facilitate accurate assay implementation and interpretation of results. The data supports that assay can aid in the management of patients with chronic HBV infection. 1. Patient Perspectives This submission did not include specific information on patient perspectives for this device. In conclusion, given the available information above, the data support that for the claimed intended use the probably benefits outweigh the probably risks. D. Overall Conclusions The data in this application support the reasonable assurance of safety and effectiveness of this device when used in accordance with the indications for use. The data from the nonclinical studies demonstrated acceptable analytical sensitivity, precision, and analytical specificity of the ADVIA Centaur Anti-HBe2 assay when used according to the instructions for use as stated in the labeling, the warnings, and precautions, and limitations sections of the labeling. The clinical studies have shown that the ADVIA Centaur Anti-HBe2 assay, when compared to the FDA approved comparator, has a similar ability to detect the presence of anti-HBe antibodies in specimens from individuals with chronic hepatitis B, or those recovered from HBV infection. The assay has also demonstrated that it has no cross-reactivity with viral antibodies or other cross-reactants in the specimens from individuals with medical conditions unrelated to the HBV infection. The probable clinical benefits outweigh the potential risks for the proposed assay considering the performance of the device in the clinical trial and the low risk and associated risk mitigations in clinical practice. The proposed assay labelling will facilitate accurate assay implementation and interpretation of results. The assay may provide substantial benefits to patients as an accurate and sensitive aid in determining HBV seroconversion in conjunction with other diagnostic information. PMA P200017: FDA Summary of Safety and Effectiveness Data Page 37 of 39 {37} # XIII. CDRH DECISION CDRH issued an approval order on July 14, 2021 Given the workforce constraints associated with the 2019 SARS-CoV-2 public health emergency, a QSIT II post-market inspection is recommended in lieu of a preapproval inspection. # XIV. APPROVAL SPECIFICATIONS - Directions for use: See device labeling. - Hazards to Health from Use of the Device: See Indications, Contraindications, Warnings, Precautions, and Adverse Events in the device labeling. - Post-approval Requirements and Restrictions: See approval order. PMA P200017: FDA Summary of Safety and Effectiveness Data Page 38 of 39
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