VITROS IMMUNODIAGNOSTIC PRODUCTS ANTI-HBE REAGENT PACK/ANTI-HBE CALIBRATOR/ANTI HBE CONTROLS

P100001 · Ortho-Clinical Diagnostics · SEI · Jul 20, 2011 · Microbiology

Device Facts

Record IDP100001
Device NameVITROS IMMUNODIAGNOSTIC PRODUCTS ANTI-HBE REAGENT PACK/ANTI-HBE CALIBRATOR/ANTI HBE CONTROLS
ApplicantOrtho-Clinical Diagnostics
Product CodeSEI · Microbiology
Decision DateJul 20, 2011
DecisionAPRL
Regulation21 CFR 866.3173
Device ClassClass 2
AttributesPediatric

Intended Use

For the in vitro qualitative detection of antibodies to hepatitis B e antigen (anti-HBe) in human adult and pediatric (2 to 21 years old) serum from individuals who have symptoms of chronic hepatitis and those who have recovered from HBV infection, using the VITROS ECi/ECiQ Immunodiagnostic Systems. Further assessment of HBV infection (biochemical, serological and/or nucleic acid testing) is required to define the specific disease state. VITROS Anti-HBe test performance has not been established for the monitoring of HBV disease or therapy.

Device Story

The VITROS Anti-HBe test is an in vitro diagnostic assay performed on the VITROS ECi/ECiQ Immunodiagnostic System. It uses a competitive chemiluminescence immunoassay technique to detect anti-HBe antibodies in human serum. The system processes samples by pre-incubating them with HBeAg, followed by the addition of biotinylated and HRP-labeled mouse monoclonal anti-HBe antibodies. The immune complex is captured on streptavidin-coated wells. A luminogenic substrate is added, and the resulting light signal is measured; the signal is inversely proportional to the anti-HBe concentration. The system automatically calculates results as a normalized signal (C/S) relative to a cutoff. Clinicians use these results alongside other serological markers to assist in the diagnosis and staging of HBV infection. The device is intended for use in clinical laboratories by trained personnel.

Clinical Evidence

Multi-center prospective study (N=1648) and a high-prevalence population study (N=208) evaluated clinical performance against an FDA-approved comparator. Positive percent agreement in chronic HBV patients was 95.92% (Pop 1) and 97.74% (Pop 2). Negative percent agreement was >99% across all groups. Pediatric performance (N=165) showed 100% concordance with the comparator. Bench testing confirmed analytical sensitivity (0.20 PEI Units/mL), specificity against 209 cross-reacting samples, and stability (40 weeks shelf-life, 12 weeks on-board).

Technological Characteristics

Competitive chemiluminescence immunoassay. Components: streptavidin-coated wells (bacterial source), rHBeAg, HRP-labeled and biotinylated mouse monoclonal anti-HBe antibodies (IgG2a, kappa). Detection via luminol derivative and peracid salt. Automated analysis on VITROS ECi/ECiQ systems. Calibration via negative Reference Calibrator. Lyophilized controls. No specific ASTM standards cited for materials.

Indications for Use

Indicated for in vitro qualitative detection of anti-HBe antibodies in human serum from adults and pediatric patients (2-21 years) with symptoms of chronic hepatitis or history of recovered HBV infection. Not for monitoring HBV disease or therapy.

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.

Reference Devices

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) Antibody to Hepatitis B e Antigen Controls Device Trade Name: VITROS Immunodiagnostic Products Anti-HBe Test (VITROS Anti-HBe) VITROS Immunodiagnostic Products Anti-HBe Reagent Pack VITROS Immunodiagnostic Products Anti-HBe Calibrator VITROS Immunodiagnostic Products Anti-HBe Controls Applicant’s Name and Address: Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, New York 14626-5101 Date of Panel Recommendation: None Premarket Approval Application (PMA) Number: P100001 Date of FDA Notice of Approval: July 20, 2011 Expedited: Not applicable. II. INDICATIONS FOR USE VITROS Immunodiagnostic Products Anti-HBe Reagent Pack For the in vitro qualitative detection of antibodies to hepatitis B e antigen (anti-HBe) in human adult and pediatric (2 to 21 years old) serum from individuals who have symptoms of chronic hepatitis and those who have recovered from HBV infection, using the VITROS ECi/ECiQ Immunodiagnostic Systems. Further assessment of HBV infection (biochemical, serological and/or nucleic acid testing) is required to define the specific disease state. VITROS Anti-HBe test performance has not been established for the monitoring of HBV disease or therapy. PMA P100001: FDA Summary of Safety and Effectiveness Data {1} VITROS Immunodiagnostic Products Anti-HBe Calibrator For use in the calibration of the VITROS ECi/ECiQ Immunodiagnostic Systems when used with the VITROS Anti-HBe test for the in vitro qualitative detection of antibodies to hepatitis B e antigen (anti-HBe). VITROS Immunodiagnostic Products Anti-HBe Controls For use in monitoring the performance of the VITROS Anti-HBe test when used on the VITROS ECi/ECiQ Immunodiagnostic Systems. ## III. CONTRAINDICATIONS None. ## IV. WARNINGS AND PRECAUTIONS The warnings and precautions can be found in the VITROS Immunodiagnostic Products Anti-HBe Reagent Pack, Calibrator and Controls labeling. ## V. DEVICE DESCRIPTION ### A. Assay Principle The VITROS Anti-HBe assay is performed using the VITROS Anti-HBe Reagent Pack and the VITROS Anti-HBe Calibrator on the VITROS ECi/ECiQ Immunodiagnostic System. The VITROS ECi/ECiQ Immunodiagnostic System allows for the determination of analytes in human samples utilizing an enhanced chemiluminescence detection reaction. A competitive assay technique is used which involves pre-incubation of anti-HBe in the sample with a fixed weight of HBeAg in the Assay Reagent, followed by incubation with a Conjugate Reagent that contains biotinylated mouse monoclonal anti-HBe antibody and horseradish peroxidase (HRP)-labeled mouse monoclonal anti-HBe antibody. The formed immune complex is captured by streptavidin on the reaction wells; unbound materials are removed by washing. The bound HRP conjugate is measured by a luminescent reaction. A reagent containing luminogenic substrates (a luminol derivative and a peracid salt) and an electron transfer agent is added to the wells. The HRP in the bound conjugate catalyzes the oxidation of the luminol derivative, producing light. The electron transfer agent (a substituted acetanilide) increases the level of light produced and prolongs its emission. The light signals are read by the VITROS ECi/ECiQ Immunodiagnostic System. The amount of PMA P100001: FDA Summary of Safety and Effectiveness Data {2} HRP conjugate bound is indirectly proportional to the concentration of anti-HBe present in the sample. B. Kit Configuration and Components a) The VITROS Immunodiagnostic Products Anti-HBe test is comprised of the following: - VITROS ECi/ECiQ Immunodiagnostic System – dedicated instrumentation, cleared by the FDA as an Immunodiagnostic System (K962919/S1), which provides automated analysis of the VITROS Immunodiagnostic System assays. - VITROS Immunodiagnostic Products Anti-HBe Reagent Pack (VITROS Anti-HBe Reagent Pack) and VITROS Immunodiagnostic Products Anti-HBe Calibrator (VITROS Anti-HBe Calibrator) together comprise the VITROS Anti-HBe assay. - VITROS Immunodiagnostic Products Anti-HBe Controls are controls used for monitoring the VITROS Anti-HBe assay. - VITROS Immunodiagnostic Products Signal Reagent and VITROS Immunodiagnostic Products Universal Wash Reagent are universal reagents used in all VITROS Immunodiagnostic System assays. b) The VITROS Immunodiagnostic Products Anti-HBe Reagent Pack is composed of three reagents: - 100 coated wells (streptavidin source, bacterial; binds ≥3ng biotin/well) - 8.4 mL assay reagent containing treated rHBeAg (4.4 Units* /mL) in buffer with mouse serum and antimicrobial agent - 6.6 mL conjugate reagent (source, HRP-mouse monoclonal anti-HBe 0.3 μg/mL and biotin-mouse monoclonal anti-HBe 5.0 μg/mL) in buffer with sheep and mouse serum and antimicrobial agent *Paul-Ehrlich-Institute Reference Serum c) The VITROS Immunodiagnostic Products Anti-HBe Calibrator contains: - 3 vials of VITROS Anti-HBe Calibrator (freeze-dried, anti-HBe positive human plasma in HBeAg and anti-HBe negative defibrinated and delipidized human plasma with antimicrobial agent); reconstitution volume 1.0 mL d) The VITROS Immunodiagnostic Products Anti-HBe Controls contains: - 3 sets of Controls 1 and 2 (freeze-dried defibrinated and delipidized human plasma with antimicrobial agent), reconstitution volume 1.0 mL. PMA P100001: FDA Summary of Safety and Effectiveness Data {3} # C. Calibration The VITROS Anti-HBe test is calibrated using a negative Reference Calibrator which is used to establish a cutoff that has optimum clinical sensitivity and specificity for the assay. The cutoff signal level is assigned a result of 1.00. The assay cutoff is maintained relative to the Reference Calibrator signal and is used to establish a valid calibration specific to each lot of reagents. Patient sample results are calculated as a normalized signal, relative to the cutoff signal (C/S): Result = Cutoff Signal / Sample Signal Assay results ≥1.00 C/S indicate a reactive sample, positive for anti-HBe. Assay results &lt;1.00 C/S indicate a non-reactive sample, negative for anti-HBe. A retest zone between ≥0.80 and &lt;1.20 requires that samples with an initial C/S value within that range to be re-tested in duplicate. # D. Interpretation of results Results are automatically calculated by the VITROS ECi/ECiQ Immunodiagnostic Systems and patient sample results are displayed with a “Negative”, “Retest?” or “Reactive” label. Below is the final result interpretation algorithm: | Initial Test Results (C/S) | Retest | Final Interpretation | | --- | --- | --- | | <0.80 | Not Required | 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 (+) | # E. Quality Control The performance of the VITROS Anti-HBe test is monitored using the VITROS Anti-HBe Controls. The performance of the VITROS Anti-HBe Controls has not been established with any other anti-HBe assay. PMA P100001: FDA Summary of Safety and Effectiveness Data {4} # 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 by using other commercially available FDA approved serological tests. This assay is one of several hepatitis marker assays that are often used together and in conjunction with clinical assessment and other laboratory test results in the diagnosis of the HBV infection. # VII. MARKETING HISTORY The VITROS Immunodiagnostic Products Anti-HBe Reagent Pack, Calibrator and Controls are currently marketed in Europe, Asia, North and South America, and Oceania. The device has not been withdrawn to date from the market in any country for reasons relating to the safety and effectiveness of the device. The following table provides the list of countries where the product is distributed currently: | Argentina | Kuwait | | --- | --- | | Armenia | Kyrgystan | | Australia | Latvia | | Austria | Lebanon | | Azerbaijan | Liberia | | Bangladesh | Libya | | Belarus | Lichtenstein | | Belguim | Lithuania | | Bolivia | Luxembourg | | Brazil | Malaysia | | Brunei | Maldives | | Bulgaria | Malta | | Burma | Martinique | | Canada | Mexico | | Chile | Moldova | | China | Nepal | | Colombia | Netherlands | | Costa Rica | New Zealand | | Croatia | Nicaragua | | Cyprus | Nigeria | | Czech Republic | Norway | | Denmark | Oman | | Dominican Republic | Panama | | Ecuador | Paraguay | | Egypt | Peru | | El Salvador | Philippines | | Estonia | Poland | | Finland | Portugal | | France | Reunion | | French Antilles | Romania | | French Guayana | Russia | | Georgia | Saudia Arabia | | Germany | Singapore | PMA P100001: FDA Summary of Safety and Effectiveness Data {5} | Greece | Slovak Republic | | --- | --- | | Guadeloupe | Slovenia | | Guatemala | South Africa | | Haiti | Spain | | Honduras | Sri Lanka | | Hong Kong | Sweden | | Hungary | Switzerland | | Iceland | Taiwan | | India | Tajikistan | | Indonesia | Thailand | | Iran | Trinidad and Tobago | | Iraq | Turkey | | Ireland | Turkmenistan | | Israel | United Arab Emirates | | Italy | United Kingdom | | Jamaica | Uruguay | | Japan | Venezuela | | Korea | Vietnam | ## VIII. POTENTIAL ADVERSE EFFECTS OF THE DEVICE ON HEALTH Below is a list of the potential adverse effects associated with the use of the device. Since the VITROS Immunodiagnostic Products Anti-HBe assay is for *in vitro* diagnostic use, there is no direct adverse effect on the health of the patient. However, 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. The risk of incorrect test results is inherent with all *in vitro* diagnostic products. Therefore, the above potential risks are not unusual in the laboratory setting and should be evaluated in conjunction with other clinical indicators. 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. Standard good laboratory practices are considered sufficient to minimize risks to the end user. ## IX. SUMMARY OF PRECLINICAL STUDIES ### A. Analytical Sensitivity The sensitivity of the VITROS Anti-HBe assay was assessed by testing a series of dilutions of a Paul-Ehrlich Institute (PEI) Anti-HBe reference serum having a known concentration. A stock solution with an anti-HBe level of 100 PEI U/mL was used to prepare an 11 member dilution series at concentrations ranging from 1.00 to 0.00 PEI U/mL. Each dilution was analyzed at n=3 using two Master Lots of reagents on two VITROS ECi/ECiQ Systems. A linear regression of the mean VITROS Anti-HBe assay PMA P100001: FDA Summary of Safety and Effectiveness Data {6} result versus the calculated concentration of each dilution was used to determine the concentration of the cut-off. The concentration at the cut-off (C/S = 1.00) of the VITROS Anti-HBe assay, as determined from the Linear Regression, was 0.20 PEI Units/mL. ## B. Cut-off Determination The position of the assay cutoff was initially based on experimental data generated on clinical samples analyzed on the VITROS ECi/ECiQ Immunodiagnostic System. The samples came from known anti-HBe reactive, seroconversion panels, blood donor samples determined to be negative for anti-HBe using other commercially available methods and clinical samples from routine laboratory testing of patients with diseases clinically related to HBV infection but expected to be negative for anti-HBe. The cut-off signal was then established as that light signal which gave the best discrimination between anti-HBe reactive and anti-HBe negative sample populations, to provide optimum specificity and sensitivity for the assay. The cut-off signal level was assigned a result of 1.00. An assay result ≥ 1.00 C/S indicates a reactive sample, positive for anti-HBe. An assay result &lt; 1.00 C/S indicates a non-reactive sample, negative for anti-HBe. ## C. Antibody Characterization The physio-chemical properties of the purified mouse monoclonal anti-HBe antibodies utilized in the VITROS Anti-HBe assay were characterized by isotype determination, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS PAGE), and isoelectric focusing (IEF) polyacrylamide gel electrophoresis. The investigation of the three lots of mouse monoclonal anti-human IgG antibodies ZAG127 (conjugated to horseradish peroxidase) and of ZAS129 (conjugated to biotin) determined that: - The isotype of the three lots of ZAG127 and of ZAS129 was IgG2a, kappa. - The SDS PAGE of the three lots of ZAG127 and of ZAS129 showed similar banding patterns with molecular weights typical of immunoglobulin heavy and light chains. - The IEF gel electrophoresis of the three lots of ZAG127 and of ZAS129 showed similar banding patterns within part number, with pI values typical of monoclonal antibodies. ## D. Potentially Cross Reacting Subgroups The specificity of the VITROS Anti-HBe assay was evaluated by testing a total of 209 patient samples representing various potentially cross-reacting sub-groups: Hepatitis C Virus (HCV), Cytomegalovirus (CMV), Epstein Barr Virus (EBV), Herpes Simplex Virus (HSV), Rubella, Syphilis, Toxoplasmosis, elevated liver enzymes (ALT), Rheumatoid Factor (RF), anti-nuclear antibodies (ANA), heterophilic anti-mouse antibodies (HAMA), past Hepatitis A infection/immunized (HAV Total), non-viral liver PMA P100001: FDA Summary of Safety and Effectiveness Data {7} disease, Autoimmune Disease (RA) Rheumatoid Arthritis, Autoimmune Disease (SLE), Parvovirus B19 infection, HIV 1/2, HTLV 1/2, recent Influenza vaccine recipients, cord blood, and E.Coli. The majority of the samples tested in this analytical specificity study were characterized based on the relevant antibody markers; documented clinical diagnosis was used to characterize the samples from the non-viral liver disease, SLE and Autoimmune Disease (RA) Rheumatoid Arthritis sub-groups. The samples were also tested in the FDA approved comparator anti-HBe assay. Of the 209 samples, 17 were found to be repeatedly reactive in the VITROS Anti-HBe assay. All 17 repeat reactive samples were also positive in the FDA approved comparator anti-HBe assay. | Sample Category | No. of Samples Tested | No. Negative | No. Initial Reactive | No. Repeat Reactive | | --- | --- | --- | --- | --- | | Hepatitis C Virus Infection (HCV) | 10 | 8 | 2 | 2* | | Cytomegalovirus (CMV) | 10 | 10 | 0 | 0 | | Epstein Barr Virus (EBV) | 10 | 10 | 0 | 0 | | Herpes Simplex Virus (HSV) | 10 | 10 | 0 | 0 | | Rubella | 10 | 10 | 0 | 0 | | Syphilis | 10 | 8 | 2 | 2* | | Toxoplasmosis | 10 | 9 | 1 | 1* | | Elevated Liver Enzymes (ALT) | 10 | 10 | 0 | 0 | | Rheumatoid Factor (RF) | 10 | 9 | 1 | 1* | | Anti-Nuclear Antibodies(ANA) | 10 | 10 | 0 | 0 | | Heterophilic anti-mouse antibodies (HAMA) | 9 | 8 | 1 | 1* | | Past Hepatitis A Infection/Immunized (HAV Total) | 10 | 8 | 2 | 2* | | Non-viral Liver Disease | 10 | 9 | 1 | 1* | | AutoImmune Disease Rheumatoid Arthritis (RA) | 10 | 10 | 0 | 0 | | AutoImmune Disease (SLE) | 10 | 9 | 1 | 1* | | Parvovirus B19 Infection | 10 | 6 | 4 | 4* | | HIV 1/2 | 10 | 8 | 2 | 2* | | HTLV 1/2 | 10 | 10 | 0 | 0 | | Recent Influenza Vaccine Recipients | 10 | 10 | 0 | 0 | | Cord Blood | 10 | 10 | 0 | 0 | | Escherichia coli | 10 | 10 | 0 | 0 | * Also positive in FDA approved comparator assay ## E. Interfering Substances The VITROS Anti-HBe assay was evaluated for interference from common endogenous substances by testing bilirubin, triolein, biotin, dipyrone, and hemoglobin, as recommended by the Clinical and Laboratory Standards Institute document EP7-A2. All interfering substance evaluations were performed using patient samples. Performance was assessed in both negative and positive samples for anti-HBe. Samples were tested in triplicate using two master lots of reagents on two VITROS ECi/ECiQ Immunodiagnostic Systems. Bilirubin, triolein, biotin and dipyrone were found not to interfere with the assay up to the concentrations indicated below: PMA P100001: FDA Summary of Safety and Effectiveness Data {8} | Compound | Concentration | | | --- | --- | --- | | Bilirubin | 0.35 mmol/L | 20 mg/dL | | Biotin | 40.8 nmol/L | 1 μg/dL | | Triolein | 33.9 mmol/L | 3000 mg/dL | | Dipyrone | 3.0 mmol/L | 100 mg/dL | Hemoglobin, when tested, was found to interfere with the VITROS Anti-HBe results. At concentrations &gt; 125 mg/dL the observed bias in the results was -26.9%. | Interferent | Interferent Concentration | Units = C/S | | | | --- | --- | --- | --- | --- | | | | | Mean Result* | Bias** | | Hemoglobin | 0.076 mmol/L | 125 mg/dL | 1.39*** | -26.9% | *Mean result of replicate determinations using 2 different lots of reagent **Estimate of the average difference observed ***Reactive sample showed negative bias The effect of elevated serum protein levels on the VITROS Anti-HBe test was not evaluated. Each clinical laboratory should verify the performance of this test with samples with high protein content in accordance with CLSI document EP7-A2. ## F. Serum Sample Stability The effect of temperature on the integrity of the anti-HBe antibody in serum samples was evaluated at 2-8 °C, at -20 °C and at room temperature (21 °C and 30 °C). Ten (10) blood samples from 10 anti-HBe negative patients were spiked with anti-HBe positive plasma to a level close to the cut-off (C/S = 2.00 +/- 1.00). Ten (10) samples were not spiked. The blood was aliquoted into serum collection tubes, centrifuged and the serum was separated from the cells. One (1) mL aliquots of each serum sample were placed at -20 °C, 2-8 °C, 21 °C and 30 °C. All samples were tested fresh and at defined time intervals during storage. Each sample was analyzed at n=3 with three Master lots of reagents using 3 instruments. The mean value for each storage condition and the overall mean across all storage conditions was calculated from the three determinations. The SD and CV(%) for anti-HBe-spiked samples were also calculated. Assay results &lt; 1.00 C/S are classified negative. Assay results ≥ 1.00 C/S are classified reactive. For each storage condition, % differences were calculated from the fresh condition for anti-HBe-spiked samples using the following equation: $$ \% \text{ Difference} = \text{Test Condition} - \text{Baseline Condition} \times 100\% \text{ Baseline Condition} $$ The mean and range of the % differences, across all anti-HBe-spiked samples, for each condition were calculated. The acceptance criteria were that no sample should be misclassified (i.e. gives a negative result rather than a reactive result) on any of the occasions on which they are determined. The acceptance criterion for the reactive samples (C/S ≥ 1.00) is that no sample should be PMA P100001: FDA Summary of Safety and Effectiveness Data {9} misclassified (i.e. gives a negative result rather than a reactive result) on any of the occasions on which they are determined. Maximum differences (change) in a mean result of + 50% and -33% would not cause a change in classification across the retest zone of 0.80 to 1.20 (C/S), therefore all % differences must fall within this range. For positive samples, no negative results were observed and for negative samples, no positive results were observed, for the mean results for any storage condition. The studies supported stability of serum samples for 5 days at 2-8°C, for 4 weeks at -20°C and 10 hours at room temperature (up to 30 °C). The effect of multiple freeze-thaw cycles on the stability of the anti-HBe in serum was also evaluated. Two serum samples reactive for anti-HBe were thawed and divided into six separate aliquots. One aliquot was stored at 2-8 °C and was considered the initial test sample while the other five were re-frozen at -20°C and subjected to five freeze/thaw cycles. Each aliquot was tested in duplicate, using two Master lots of reagents on two instruments. Same acceptance criteria, as described above, were applied. The data showed that up to five (5) freeze thaw cycles had no effect on the results of the VITROS Anti-HBe assay. ## G. Reagent Stability ### 1. VITROS Anti-HBe Reagent Pack and Calibrator #### a. Long Term Stability (Shelf Life) VITROS Anti-HBe Reagent Packs and Calibrators that were subjected to a period of simulated transport by storage at 20°C for 2 days, and then returned to 2-8°C, were tested at monthly intervals up to 56 weeks using in-house controls and VITROS Anti-HBe Controls. Four runs were performed on 3 Master Lots of the VITROS Anti-HBe Reagent Pack and Calibrator at each timepoint. Each run contained duplicate determinations of the VITROS Anti-HBe Calibrator and singleton determinations of the QC In-house Controls (four levels) and the VITROS Anti-HBe Controls. The final shelf-life was defined as the time point where all parameters for all Master Lots pass the acceptability limits prior to the time points at which any same parameter for a Master Lot has failed the acceptance limits on two successive occasions. In addition, at initial, interim and expiry time points, runs were performed using a performance panel obtained from Boston Biomedica Inc. to assess the seroconversion sensitivity of the VITROS Anti-HBe assay throughout the shelf life. The results indicated that the performance panel samples retained their classification at the interim and expiry time points for all lots of material evaluated up to 52 weeks. The data supported the claimed shelf life stability of 40 weeks for the VITROS Anti-HBe Reagent Pack and Calibrator. PMA P100001: FDA Summary of Safety and Effectiveness Data page 10 14 {10} b. Temperature Stressing at -20 °C VITROS Anti-HBe Reagent Packs and Calibrator were evaluated for effects of extreme temperature variation on their performance by exposing the reagents to freezing. All the reagents tested were subjected to a period of simulated transport (stored at 20°C for 2 days and then returned to 2-8°), prior to initiating the stability study, to mimic the effects of shipment. To evaluate the effect of freezing, three Master Lots of the VITROS Anti-HBe Reagent Packs and Calibrator were subjected to two freeze/thaw cycles and the performance was compared with Reagent Packs and Calibrators stored at 2-8 °C (unstressed). There were four runs performed for each combination of Reagent Packs and Calibrators. Each run consisted of duplicate determinations of the Calibrator and single determinations of the in-house Controls and the VITROS Anti-HBe Controls as follows: | Unstressed Reagent Pack and Unstressed Calibrator | | --- | | Unstressed Reagent Pack and Stressed Calibrator | | Stressed Reagent Pack and Unstressed Calibrator | | Stressed Reagent Pack and Stressed Calibrator | The study results demonstrated that inadvertent freezing of the Reagent Pack and/or Calibrator had no adverse effect on Calibration Quality Parameters or Control results. It is recommended that the VITROS Anti-HBe Reagent Pack and Calibrator are not frozen. c. Temperature Stressing at 30°C and 37°C VITROS Anti-HBe Reagent Packs and Calibrator were evaluated for effects of extreme temperature variation on their performance by exposing the reagents to 30°C and 37°C. All the reagents tested were subjected to a period of simulated transport, prior to initiating the stability study, to mimic the effects of shipment. VITROS Anti-HBe Reagent Packs and Calibrators from three Master Lots were stored for 5 days at 30 °C (86 °F) or for 1 day at 37 °C (98.6 °F). Four runs were performed for each combination of Reagent Packs and Calibrators. Each run consisted of duplicate determinations of the Calibrator and single determinations of the in-house Controls and the VITROS Anti-HBe Controls as follows: | Unstressed Reagent Pack and Unstressed Calibrator | | --- | | Unstressed Reagent Pack and Stressed Calibrator | | Stressed Reagent Pack and Unstressed Calibrator | | Stressed Reagent Pack and Stressed Calibrator | PMA P100001: FDA Summary of Safety and Effectiveness Data page 11 15 {11} The results showed that exposing the VITROS Anti-HBe Reagent Packs to a temperature of $30^{\circ}\mathrm{C}$ ($86^{\circ}\mathrm{F}$) for 5 days or $37^{\circ}\mathrm{C}$ ($98.6^{\circ}\mathrm{F}$) for 1 day caused detrimental effects on Calibrator Signal Index results. Inadvertent exposure of the Reagent Packs to these temperatures for the times stated would significantly compromise the performance of the VITROS Anti-HBe assay. However, inadvertent storage of the VITROS Anti-HBe Calibrator at $30^{\circ}\mathrm{C}$ for 5 days or $37^{\circ}\mathrm{C}$ for 1 day would not significantly compromise the performance of the VITROS Anti-HBe assay. d. On Board Stability (Open Stability) A real-time open Reagent Pack stability study was conducted using 3 Master Lots of Reagent Pack and Calibrator and aged generic reagents (Signal Reagent, Universal Wash Reagent) to assess the effect of open on board storage of the VITROS Anti-HBe Reagent Pack for a period of 12 weeks. All the reagents tested were subjected to a period of simulated transport (stored at $20^{\circ}\mathrm{C}$ for 2 days and then returned to $2-8^{\circ}\mathrm{C}$ prior to testing), prior to initiating the stability study, to mimic the effects of shipment. The VITROS Anti-HBe Reagent Packs and Calibrators that were subjected to a period of simulated transport, were opened and placed in an environmental chamber ($4-8^{\circ}\mathrm{C}$ at $\leq 40\%$ relative humidity) for a period of up to 12 weeks to simulate the storage of the reagent packs on board the VITROS ECi/ECiQ System. Reagent packs and calibrators were removed from the chamber at 2 week intervals and used to test in-house controls and VITROS Anti-HBe Controls, performing four runs for each Master Lot. Each run contained duplicate determinations of the Calibrator and single determinations of the QC In-house Controls. The data demonstrated that VITROS Anti-HBe Reagent Packs and Calibrators can be stored open, on board the VITROS ECi/ECiQ System for a period of up to 12 weeks. e. Calibrator Open Stability VITROS Anti-HBe Calibrators were reconstituted, pooled, transferred to sample cups and then stored at $2-8^{\circ}\mathrm{C}$ and $-20^{\circ}\mathrm{C}$ for up to 13 weeks. The stored Calibrators were compared against fresh Calibrators at various time points throughout the 13 weeks using three Master Lots. The results at each time point indicated that there were no differences between the fresh and stored Calibrators, and no trends throughout the duration of storage were evident. The data supports the storage of the VITROS Anti-HBe Calibrators, after reconstitution, for a period of up to 13 weeks at $2-8^{\circ}\mathrm{C}$, or up to 13 weeks at $-20^{\circ}\mathrm{C}$. PMA P100001: FDA Summary of Safety and Effectiveness Data {12} # 2. VITROS Anti-HBe Controls Stability ## a. Long Term Stability (Shelf Life) VITROS Anti-HBe Controls stability study was conducted using two lots of VITROS Anti-HBe Controls, three Master Lots of assay reagents (Reagent Pack/Calibrator) and aged generic reagents (Signal Reagent and Universal Wash Reagent). VITROS Anti-HBe Reagent Packs, Calibrators and Controls that were subjected to a period of simulated transport by storage at $20^{\circ}\mathrm{C}$ for 2 days, and then returned to $2-8^{\circ}\mathrm{C}$, were tested at monthly intervals up to 56 weeks. Prior to the commencement of the stability study, results obtained from transported materials were compared to results obtained from non-transported materials to verify that QC In-house Controls results were not affected; thereby confirming that assay performance is maintained. Four runs were performed on 3 Master Lots of the VITROS Anti-HBe Reagent Pack and Calibrator at each timepoint. Each run contained duplicate determinations of the VITROS Anti-HBe Calibrator and singleton determinations of the QC In-house Controls (four levels) and the VITROS Anti-HBe Controls. The study data support a shelf life stability for lyophilized VITROS Anti-HBe Controls of 52 weeks when stored at $2-8^{\circ}\mathrm{C}$. ## b. Reconstituted stability Two lots of the VITROS Anti-HBe Controls were evaluated for stability after reconstitution of the lyophilized material. The testing was done with three Master Lots of the Reagent Pack and Calibrator) and aged generic reagents (Signal Reagent and Universal Wash Reagent). The Reagent Packs, Calibrators and the VITROS Anti-HBe Controls were subjected to simulated transport conditions (stored at $20^{\circ}\mathrm{C}$ for 2 days and then returned to $2-8^{\circ}\mathrm{C}$ prior to the commencement of the stability study). The results obtained from transported materials were compared to results obtained from non-transported materials to verify that QC In-house Controls results were not affected; thereby confirming that assay performance is maintained. The VITROS Anti-HBe Controls were reconstituted, pooled and stored in sample cups at $2-8^{\circ}\mathrm{C}$ and at $-20^{\circ}\mathrm{C}$. The pooled Controls were tested on the day of reconstitution as time point 0. The VITROS Anti-HBe Controls stored at $2-8^{\circ}\mathrm{C}$ were subsequently tested on days 3, 4, 5 and 7. The VITROS Anti-HBe Controls stored at $-20^{\circ}\mathrm{C}$ and thawed one time were tested on weeks 1, 2, 3 and 4 of the trial. Additionally, the VITROS Anti-HBe Controls stored at $-20^{\circ}\mathrm{C}$ were subjected to 3 Freeze/Thaw cycles and were tested on week 4 of the trial. Four runs were performed at all time points using each Master Lot of Reagent Pack and Calibrator. Each run contained duplicate determinations of the Calibrator and PMA P100001: FDA Summary of Safety and Effectiveness Data page 13 17 {13} singleton determinations of the QC In-house Controls and VITROS Anti-HBe Controls. All results met the predetermined acceptance criteria. The performance observed was comparable between the freshly reconstituted VITROS Anti-HBe Controls and the reconstituted VITROS Anti-HBe Controls stored at 2-8°C, -20°C and -20°C with 3 Freeze/Thaw cycles. The data supports the stability claim of 5 days storage at 2-8°C or up to 4 weeks storage at -20°C with 3 Freeze/Thaw cycles for the VITROS Anti-HBe Controls after reconstitution. ## 3. Universal Wash Reagent VITROS Anti-HBe Reagent Packs, Calibrators and Controls that were subjected to a period of simulated transport to mimic the effects of shipment were tested with three lots of VITROS Universal Wash Reagent at 0, 6 and 12 months of age to determine the effect of aged VITROS Universal Wash Reagent on VITROS Anti-HBe results. The data indicated that the performance of the VITROS Anti-HBe assay is acceptable when used with VITROS Universal Wash Reagent which is either fresh, 6 or 12 months old. ## 4. Signal Reagent Stability VITROS Anti-HBe Reagent Packs, Calibrator and Controls that were subjected to a period of simulated transport to mimic the effects of shipment were tested with four lots of VITROS Signal Reagent stored at 2-8 °C for up to 6 months to determine the effect of aged VITROS Signal Reagent on VITROS Anti-HBe results. The data indicated that the performance of the VITROS Anti-HBe assay is acceptable when used with VITROS Signal Reagent which is either fresh, or 6 months old. ## 5. Preservative Effectiveness Three aspects of microbiological control were studied in VITROS Anti-HBe Reagent Pack and Calibrator reagents: - Determination of post-dispensing microbial load at 52 weeks - Preservative concentration over a 52 week shelf-life - Preservative efficacy 52 weeks post formulation Results of the studies indicated that the level of the preservative used in the reagents was adequate for microbial control over the 52 week time period. ## H. Seroconversion Sensitivity PMA P100001: FDA Summary of Safety and Effectiveness Data page 14 18 {14} The clinical sensitivity of the VITROS Anti-HBe assay was evaluated by testing six commercially available seroconversion panels. The VITROS Anti-HBe and FDA approved comparator anti-HBe test results are summarized below. The table presents the days elapsed from the date of the initial bleed for the last negative sample and first repeatedly reactive sample for the VITROS Anti-HBe and for the FDA approved comparator anti-HBe test as well as the difference between the two tests in identifying the first reactive panel member by number of days. | Days to Reactive anti-HBe Result | | | | | | | --- | --- | --- | --- | --- | --- | | Panel ID | Comparator Anti-HBe Test | | VITROS Anti-HBe Test | | Difference in Days to Anti-HBe Reactive Result | | | -* | +** | -* | +** | Comparator minus VITROS Anti-HBe Test | | PHM935B | 61 | 75 | 103 | 118 | -43 | | RP009 | 81 | 88 | 81 | 88 | 0 | | RP016 | 59 | 73 | 73 | 78 | -5 | | RP017 | 78 | 140 | 78 | 140 | 0 | | 6510 | 56 | 70 | 56 | 70 | 0 | | 6513 | 98 | 112 | 98 | 112 | 0 | * Post bleed day of last negative result, usually denotes previous bleed from first positive/reactive result. **Post bleed day of first positive/reactive result. ## I. Calibration Interval The performance of the VITROS Anti-HBe assay within and beyond one calibration interval was evaluated in conjunction with the precision study (see below) which was conducted within one calibration interval (28 days). The study utilized a six member panel with C/S values around the cutoff, ranging from 0.40 to 1.56 C/S. Additional testing was conducted on days 29 and 30 to show that the analyzer would still yield valid results beyond the end of the 28 day cycle. The least squares regression analyses were performed within site and across sites. The VITROS Anti-HBe assay demonstrated adequate performance throughout the calibration interval (28 days) and continued to perform successfully two days beyond the expiration of calibration. ## J. Precision The precision study was performed at three clinical testing sites over 28 days, using one lot of reagents. The mean C/S for the six panel members ranged from 0.38 to 1.60 for the total of 120 observations (3 sites). Each testing day included one replicate of Control 1 and Control 2, followed by two replicates of each panel member. A total of 40 observations were generated at each site for each panel member. The mean of the C/S ratio, relative variance, SD and %CV were calculated and presented for each site separately and overall. The data presented are a representation of the product performance, and were rounded following all calculations. The results are shown below. PMA P100001: FDA Summary of Safety and Effectiveness Data {15} | Site | No. of Obs. | No. of Days | Mean VITROS Anti-HBe Test C/S Ratio | Precision | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | | Within Day* | | Between Days** | | Total*** | | | | | | SD | CV (%) | SD | CV (%) | SD | CV (%) | | | Site 1 | 40 | 20 | 1.54 | 0.038 | 2.5 | 0.092 | 6.0 | 0.100 | 6.5 | | | 40 | 20 | 1.41 | 0.043 | 3.1 | 0.070 | 5.0 | 0.083 | 5.9 | | | 40 | 20 | 1.27 | 0.041 | 3.2 | 0.075 | 5.9 | 0.085 | 6.7 | | | 40 | 20 | 0.80 | 0.021 | 2.6 | 0.036 | 4.5 | 0.042 | 5.2 | | | 40 | 20 | 0.63 | 0.012 | 1.8 | 0.031 | 4.9 | 0.033 | 5.2 | | | 40 | 20 | 0.40 | 0.007 | 1.6 | 0.018 | 4.6 | 0.020 | 4.9 | | Site 2 | 40 | 20 | 1.52 | 0.043 | 2.9 | 0.111 | 7.3 | 0.119 | 7.9 | | | 40 | 20 | 1.39 | 0.040 | 2.8 | 0.100 | 7.2 | 0.107 | 7.7 | | | 40 | 20 | 1.22 | 0.045 | 3.7 | 0.090 | 7.4 | 0.101 | 8.3 | | | 40 | 20 | 0.77 | 0.025 | 3.3 | 0.044 | 5.7 | 0.051 | 6.6 | | | 40 | 20 | 0.61 | 0.013 | 2.2 | 0.039 | 6.4 | 0.041 | 6.7 | | | 40 | 20 | 0.38 | 0.008 | 2.0 | 0.024 | 6.2 | 0.025 | 6.5 | | Site 3 | 40 | 20 | 1.60 | 0.060 | 3.7 | 0.125 | 7.8 | 0.138 | 8.6 | | | 40 | 20 | 1.49 | 0.041 | 2.8 | 0.147 | 9.9 | 0.153 | 10.2 | | | 40 | 20 | 1.31 | 0.031 | 2.4 | 0.100 | 7.6 | 0.104 | 8.0 | | | 40 | 20 | 0.84 | 0.017 | 2.1 | 0.065 | 7.8 | 0.067 | 8.0 | | | 40 | 20 | 0.65 | 0.015 | 2.4 | 0.046 | 7.1 | 0.048 | 7.4 | | | 40 | 20 | 0.41 | 0.006 | 1.4 | 0.030 | 7.2 | 0.030 | 7.4 | * Within Day: Variability of the assay performance from replicate to replicate. ** Between Days: Variability of the assay performance from day to day. *** Total: Variability of the assay performance combining the effects of within day and between days. ## K. Reproducibility Reproducibility of the assay was also evaluated incorporating between site and between lot variations. The study was performed at three external sites using three reagent lots to test three replicates each of a six member panel on a single occasion per day on six different days. The between site, between lot, and total precision estimates (CV (%)) were derived from a variance component analysis. The data shown in the table are a representation of the product performance, and were rounded following all calculations. PMA P100001: FDA Summary of Safety and Effectiveness Data page 16 20 {16} | Mean VITROS Anti-HBe Assay | Between Site * | | Between Lot ** | | Total *** | | No. of | | --- | --- | --- | --- | --- | --- | --- | --- | | C/S Ratio | SD | CV (%) | SD | CV (%) | SD | CV (%) | Obs. | | 1.41 | 0.098 | 7.0 | 0.079 | 5.6 | 0.185 | 13.2 | 162 | | 1.32 | 0.087 | 6.6 | 0.057 | 4.3 | 0.160 | 12.1 | 162 | | 1.18 | 0.067 | 5.7 | 0.054 | 4.5 | 0.141 | 11.9 | 162 | | 0.73 | 0.051 | 6.9 | 0.031 | 4.2 | 0.083 | 11.3 | 162 | | 0.57 | 0.047 | 8.2 | 0.023 | 4.0 | 0.067 | 11.8 | 162 | | 0.36 | 0.031 | 8.5 | 0.013 | 3.6 | 0.043 | 11.9 | 162 | * Between Site: Variability of the assay performance from site to site. ** Between Lot: Variability of the assay performance from lot to lot calculated using data across all sites. *** Total: Variability of the assay incorporating factors of site, lot and day. ## L. Carryover Studies Potential sources of cross-contamination on the VITROS ECi/ECiQ Immunodiagnostic System have been identified to be: 1) Sample-to-sample; 2) Reagent-to-reagent; and 3) Well wash-to-well wash. Sample-to-sample cross-contamination is mitigated by the use of disposable sample metering tips for each sample. The potential of reagent cross-contamination is of concern in particular between Anti-HBe and HBeAg assays, if run concurrently on the analyzer, since both assays utilize a labeled anti-HBe antibody as one of the critical reagents. The system was challenged in two experiments designed to detect a possible carryover of reagents. The first study utilized 100 anti-HBe negative samples concurrently analyzed with the VITROS anti-HBe and the VITROS HBeAg assays (worst case scenario), to challenge both the reagent-to-sample and reagent-to-reagent carryover effect. The data showed no drifting or spiking in the negative result for anti-HBe which demonstrated that there is no reagent-to-sample or reagent-to-reagent carryover effect from the VITROS HBeAg assay reagents. A second carryover challenge was conducted using 5 high anti-HBe positive samples as a worst case of potential sample carryover from well wash-to-well wash. A single negative sample was run at $n=10$ to establish a baseline. Next, the negative sample was placed in the tray immediately following each positive sample in the tray and the samples were processed in singleton. The experiment was repeated using a high negative sample that was created by diluting a positive sample to $\sim 0.8$ C/S. The results showed that there was no statistically significant difference in the mean of the negative sample. The mean of the negative sample results was 0.16 C/S (negative only) and 0.15 C/S (following high positive); the mean of the high negative was 0.79 C/S (negative only) and 0.77 C/S (following high positive). PMA P100001: FDA Summary of Safety and Effectiveness Data {17} The results demonstrated that there is no detectable cross-contamination that could affect the VITROS anti-HBe assay results when used on the VITROS ECi/ECiQ Immunodiagnostic System. ## X. SUMMARY OF PRIMARY CLINICAL STUDIES A multi-center prospective study was conducted to evaluate the clinical performance of the VITROS Anti-HBe test among individuals with signs or symptoms of hepatitis (i.e. fatigue, anorexia, malaise, nausea, jaundice, abdominal pain, dark urine, headache, vomiting, weight loss, hepatomegaly and elevated liver function tests) or biochemical manifestations (elevated liver function tests) of hepatitis and those at high risk of hepatitis infection due to lifestyle, behavior, occupation, or known exposure events. ### A. Study Population The prospective study population was divided into two groups. Population 1 consisted of 1976 subjects prospectively enrolled at four geographically separate locations; subjects were enrolled in Miami, FL (53.5%), in Dallas, TX (14.3%), in Newark, NJ (6.2%), and in Chicago, IL (26.0%). The group was Caucasian (18.9%), African American (51.0%), Hispanic (23.8%), and Asian (2.5%) with the remaining 3.8% represented by other ethnic groups. The group was 53.8% male and 46.2% female and ranged in age from 5 to 89 years. Population 2 consisted of 311 subjects living in an area of India with high prevalence of HBV infection. All patients in Population 2 presented with signs or symptoms of viral hepatitis. The mean age of the population was 39 years and the median age was 40 years. Approximately 87% of the study subjects were ≤50 years of age. The minimum age was 18 years and the maximum age was 90 years. The population was 27% female and 73% male. Testing of the samples with the VITROS Anti-HBe test was performed at diagnostic laboratories located in Miami, FL, Port Jefferson, NY and St. Paul, MN. All study samples were also tested with the FDA approved comparator anti-HBe assay at diagnostic laboratories located in Miami, FL, Los Angeles, CA and St. Paul, MN. ### B. HBV Disease Classification All patients were serologically characterized using a hepatitis marker profile consisting of previously FDA approved tests for the detection of HBsAg, HBeAg, anti-HBc Total, anti-HBc IgM, anti-HBe, and anti-HBs (quantitative). The FDA approved tests' procedures were adhered to during the clinical study. The following positive (+) / negative (-) patterns for the six HBV serological markers were used to assign an HBV disease classification of chronic, recovered, vaccinated and not previously infected with HBV: PMA P100001: FDA Summary of Safety and Effectiveness Data page 18 22 {18} | HBsAg* ** | HBeAg | aHBc IgM | aHBc Total | aHBe*** | aHBs*** ≥10 mlU/mL | HBV Disease Classification | | --- | --- | --- | --- | --- | --- | --- | | + | + | - | + | + | - | Chronic | | + | + | - | + | - | + | Chronic | | + | + | - | + | - | - | Chronic | | + | + | - | + | - | I | Chronic | | + | - | - | + | + | + | Chronic | | + | - | - | + | + | - | Chronic | | + | - | - | + | + | I | Chronic | | + | - | - | + | - | + | Chronic | | + | - | - | + | - | - | Chronic | | + | - | - | + | I | - | Chronic | | - | - | - | + | - | + | Recovered | | - | - | - | + | - | - | Recovered | | - | - | - | + | - | I | Recovered | | - | - | - | - | - | + | HBV Vaccine Response | | - | - | - | - | - | I | HBV Vaccine Response | | - | - | - | - | - | - | Not Previously Infected with HBV | * Positive = HBsAg test positive or reactive and confirmed by neutralization. ** Negative = HBsAg test negative or not confirmed by neutralization. *** I = Indeterminate result. ## C. Clinical performance The clinical study data were analyzed following the assignment of HBV disease classifications. The data analysis included only the individuals that had hepatitis marker profiles consistent with chronic infection, recovered, vaccinated and not previously infected with HBV. ## 1. Expected Results Of the 1976 subjects in Population 1 who were tested in the VITROS Anti-HBe clinical study, 1648 samples were derived from individuals who were chronically infected, recovered, vaccinated and those not previously infected with HBV. All 1648 were either at risk for HBV due to lifestyle, behavior, occupation, or a known exposure event or had signs and symptoms of hepatitis. Subjects in this group were enrolled in Miami, FL (51.2%), in Dallas, TX (15.0%), in Newark, NJ (6.4%), and in Chicago, IL (27.4%). The group was Caucasian (20.2%), African American (48.7%), Hispanic (25.4%), and Asian PMA P100001: FDA Summary of Safety and Effectiveness Data {19} (2.2%) with the remaining 3.5% represented by other ethnic groups. The group was 52.7% male and 47.3% female and ranged in age from 5 to 89 years. The distribution of VITROS Anti-HBe reactive and non-reactive results among the chronically infected, recovered, vaccinated and those not previously infected with HBV is presented, stratified by age and gender, in the following table. Expected Results for Study Subjects in Population 1 (N=1648) | | | Reactive | | Negative | | | | --- | --- | --- | --- | --- | --- | --- | | Age Range | Gender | N* | Percent** | N*** | Percent† | Total§ | | ≤ 15 | Female | 0 | 0.0 | 2 | 100 | 2 | | | Male | 0 | 0.0 | 4 | 100 | 4 | | 16-20 | Female | 2 | 7.1 | 26 | 92.9 | 28 | | | Male | 0 | 0.0 | 19 | 100 | 19 | | 21-30 | Female | 1 | 0.8 | 126 | 99.2 | 127 | | | Male | 1 | 0.8 | 124 | 99.2 | 125 | | 31-40 | Female | 1 | 0.6 | 165 | 99.4 | 166 | | | Male | 9 | 4.8 | 177 | 95.2 | 186 | | 41-50 | Female | 1 | 0.5 | 196 | 99.5 | 197 | | | Male | 17 | 6.3 | 252 | 93.7 | 269 | | 51-60 | Female | 3 | 1.9 | 158 | 98.1 | 161 | | | Male | 11 | 5.8 | 178 | 94.2 | 189 | | 61-70 | Female | 1 | 1.6 | 63 | 98.4 | 64 | | | Male | 1 | 1.7 | 58 | 98.3 | 59 | | > 70 | Female | 0 | 0.0 | 33 | 100 | 33 | | | Male | 1 | 5.9 | 16 | 94.1 | 17 | | Unknown | Female | 0 | 0.0 | 1 | 100 | 1 | | | Male | 0 | 0.0 | 1 | 100 | 1 | | Total | | 49 | 3.0 | 1599 | 97.0 | 1648 | * The total number (N) of subjects in each age range/gender category with reactive VITROS Anti-HBe results. ** The total number (N) of subjects in each age range/gender category that are reactive expressed as a percentage (%) of all subjects in that category. *** The total number (N) of subjects in each age range/gender category with negative VITROS Anti-HBe results. † The total number (N) of subjects in each age range/gender category that are negative expressed as a percentage (%) of all subjects in that category. § The total number (N) of subjects in each age range/gender category. PMA P100001: FDA Summary of Safety and Effectiveness Data page 20 24 {20} All subjects enrolled in Population 2 (N=311) were from an area in India with a high prevalence of HBV infection and all presented with signs or symptoms of viral hepatitis. Of the 311 subjects, 208 were from individuals who were chronically infected, recovered, vaccinated and those not previously infected with HBV. The mean age of these patients was 39 years and the median age was 40 years. Approximately 87% were &lt;50 years of age. The minimum age was 18 years and the maximum age was 90 years. The group was 32.2% female and 67.8% male. The VITROS Anti-HBe test was reactive in 62.5% (130/208) of the individuals in this group. The distribution of VITROS Anti-HBe reactive and non-reactive results among the chronically infected, recovered, vaccinated and those not previously infected with HBV is presented, stratified by age and gender, in the following table. Expected Results for Study Subjects in Population 2 (N=208) | Age Range | Gender | Reactive | | Negative | | Total§ | | --- | --- | --- | --- | --- | --- | --- | | | | N* | Percent** | N*** | Percent† | | | 18–20 | Female | 2 | 40.0 | 3 | 60.0 | 5 | | | Male | 2 | 25.0 | 6 | 75.0 | 8 | | 21–30 | Female | 16 | 61.5 | 10 | 38.5 | 26 | | | Male | 15 | 51.7 | 14 | 48.3 | 29 | | 31–40 | Female | 15 | 60.0 | 10 | 40.0 | 25 | | | Male | 22 | 53.7 | 19 | 46.3 | 41 | | 41–50 | Female | 4 | 57.1 | 3 | 42.9 | 7 | | | Male | 30 | 76.9 | 9 | 23.1 | 39 | | 51–60 | Female | 2 | 66.7 | 1 | 33.3 | 3 | | | Male | 15 | 88.2 | 2 | 11.8 | 17 | | 61–70 | Female | 1 | 100 | 0 | 0.0 | 1 | | | Male | 5 | 83.3 | 1 | 16.7 | 6 | | >70 | Male | 1 | 100 | 0 | 0.0 | 1 | | Total | | 130 | 62.5 | 78 | 37.5 | 208 | * The total number (N) of subjects in each age range/gender category with reactive VITROS Anti-HBe results. ** The total number (N) of subjects in each age range/gender category that are reactive, expressed as a percentage (%) of all subjects in that category. *** The total number (N) of subjects in each age range/gender category with negative VITROS Anti-HBe results. † The total number (N) of subjects in each age range/gender category that are negative, expressed as a percentage (%) of all subjects in that category. § The total number (N) of subjects in each age range/gender category. Expected results for the VITROS Anti-HBe test were also determined using prospective samples from a population of pediatric subjects in Florida (N=165). The group was 47.9% male and 52.1% female, and the subjects’ ages ranged from 2 through 21 years. The expected results are presented in the following table. PMA P100001: FDA Summary of Safety and Effectiveness Data {21} Expected Results for Pediatric Subjects (N=165) | Age Range | Gender | Reactive | | Negative | | Total$ | | --- | --- | --- | --- | --- | --- | --- | | | | N* | Percent** | N*** | Percent† | | | 2–4 | Female | 0 | 0.0 | 13 | 100 | 13 | | | Male | 0 | 0.0 | 11 | 100 | 11 | | 5–8 | Female | 0 | 0.0 | 18 | 100 | 18 | | | Male | 0 | 0.0 | 17 | 100 | 17 | | 9–12 | Female | 0 | 0.0 | 17 | 100 | 17 | | | Male | 0 | 0.0 | 20 | 100 | 20 | | 13–16 | Female | 0 | 0.0 | 21 | 100 | 21 | | | Male | 0 | 0.0 | 14 | 100 | 14 | | 17–21 | Female | 0 | 0.0 | 17 | 100 | 17 | | | Male | 0 | 0.0 | 17 | 100 | 17 | | Total | | 0 | 0.0 | 165 | 100 | 165 | * The total number (N) of subjects in each age range/gender category with reactive VITROS Anti-HBe results. ** The total number (N) of subjects in each age range/gender category that are reactive, expressed as a percentage (%) of all subjects in that category. *** The total number (N) of subjects in each age range/gender category with negative VITROS Anti-HBe results. † The total number (N) of subjects in each age range/gender category that are negative, expressed as a percentage (%) of all subjects in that category. § The total number (N) of subjects in each age range/gender category. ## 2. Agreement with a Comparator Assay The VITROS Anti-HBe assay performance was evaluated for positive and negative agreement with an FDA approved comparator anti-HBe assay. a) The following table compares the VITROS Anti-HBe results with the FDA approved comparator anti-HBe test results stratified by HBV disease classification for the 1648 subjects in Population 1 who were classified as chronically infected, recovered, vaccinated or not previously infected with HBV. Comparison of Anti-HBe Test Results by HBV Disease Classification in Population 1 (N=1648) | HBV Disease Classification | Comparator Anti-HBe Test Result | | | | | | Total | | --- | --- | --- | --- | --- | --- | --- | --- | | | Positive | | Negative | | Indeterminate | | | | | VITROS Anti-HBe Test Result | | VITROS Anti-HBe Test Result | | VITROS Anti-HBe Test Result | | | | | Reactive | Negative | Reactive | Negative | Reactive | Negative | | | Chronic | 47 | 1 | 0 | 21* | 0 | 1 | 70 | | Recovered | 0 | 0 | 1 | 218 | 0 | 0 | 219 | | HBV Vaccine Response | 0 | 0 | 0 | 314 | 0 | 0 | 314 | | Not Previously Infected with HBV | 0 | 0 | 1 | 1044 | 0 | 0 | 1045 | | Overall | 47 | 1 | 2 | 1597 | 0 | 1 | 1648 | * Three samples were HBeAg negative. PMA P100001: FDA Summary of Safety and Effectiveness Data {22} b) The following table compares the VITROS Anti-HBe results with the FDA approved comparator anti-HBe test results stratified by HBV disease classification for the 208 subjects in Population 2 who were classified as chronically infected, recovered, vaccinated or not previously infected with HBV. Comparison of Anti-HBe Test Results by HBV Disease Classification in Population 2 (N=208) | HBV Disease Classification | Comparator Anti-HBe Test Result | | | | | | Total | | --- | --- | --- | --- | --- | --- | --- | --- | | | Positive | | Negative | | Indeterminate | | | | | VITROS Anti-HBe Test Result | | VITROS Anti-HBe Test Result | | VITROS Anti-HBe Test Result | | | | | Reactive | Negative | Reactive | Negative | Reactive | Negative | | | Chronic | 130 | 3 | 0 | 52* | 0 | 0 | 185 | | Recovered | 0 | 0 | 0 | 3 | 0 | 0 | 3 | | HBV Vaccine Response | 0 | 0 | 0 | 3 | 0 | 0 | 3 | | Not Previously Infected with HBV | 0 | 0 | 0 | 17 | 0 | 0 | 17 | | Overall | 130 | 3 | 0 | 75 | 0 | 0 | 208 | | * One sample was HBeAg negative. | | | | | | | | ## 3. Percent Agreement Positive and negative percent agreement between the VITROS Anti-HBe test 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 VITROS Anti-HBe 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 VITROS Anti-HBe results by the total number of subjects negative with the comparator anti-HBe test. a) Population 1 The following table shows positive and negative percent agreement between the VITROS Anti-HBe test and the FDA approved comparator anti-HBe test along with the 95% exact confidence intervals for Population 1. PMA P100001: FDA Summary of Safety and Effectiveness Data {23} Positive and Negative Percent Agreement by HBV Disease Classification in Population 1 (N=1648) | HBV Disease Classification† | Positive Percent Agreement % (N/Total)* | 95% Exact Confidence Interval | Negative Percent Agreement % (N/Total)* | 95% Exact Confidence Interval | | --- | --- | --- | --- | --- | | Chronic | 95.92% (47/49) | 86.02–99.50% | 100.0% (21/21)*** | 83.89–100.0% | | Recovered | N/A (0/0)** | N/A | 99.54% (218/219) | 97.48–99.99% | | HBV Vaccine Response | N/A (0/0)** | N/A | 100.0% (314/314) | 98.83–100.0% | | Not Previously Infected with HBV | N/A (0/0)** | N/A | 99.90% (1044/1045) | 99.47–100.0% | * VITROS Anti-HBe negative/ comparator indeterminate results (N=1) were considered VITROS Anti-HBe false negative when calculating positive agreement. ** There were no subjects with this HBV disease classification whose comparator test results fell within this category. *** Three samples were HBeAg negative. † No standalone anti-HBe test is adequate to stage HBV disease. The positive percent agreement in samples with a chronic HBV disease classification in Population 1 was 95.92%. One VITROS Anti-HBe negative sample that was "indeterminate" by the comparator test was considered VITROS Anti-HBe false negative when calculating positive agreement. The negative percent agreement was 100.0%. b) Population 2 Positive and negative percent agreement between the VITROS Anti-HBe test and the comparator anti-HBe test were calculated for subjects in Population 2. The following table summarizes these calculations and provides the 95% exact confidence intervals for this group. Positive and Negative Percent Agreement by HBV Disease Classification in Population 2 (N=208) | HBV Disease Classification† | Positive Percent Agreement % (N/Total) | 95% Exact Confidence Interval | Negative Percent Agreement % (N/Total) | 95% Exact Confidence Interval | | --- | --- | --- | --- | --- | | Chronic | 97.74% (130/133) | 93.55–99.53% | 100.0% (52/52)*** | 93.15–100.0% | | Recovered | N/A (0/0)** | N/A | 100.0% (3/3) | N/A* | | HBV Vaccine Response | N/A (0/0)** | N/A | 100.0% (3/3) | N/A* | | Not Previously Infected with HBV | N/A (0/0)** | N/A | 100.0% (17/17) | 80.49–100.0% | * Confidence intervals calculated on small numbers are not meaningful. ** There were no subjects with this HBV disease classification whose comparator test results fell within this category. *** One sample was HBeAg negative. † No standalone anti-HBe test is adequate to stage HBV disease. PMA P100001: FDA Summary of Safety and Effectiveness Data {24} The positive percent agreement in samples with chronic HBV disease classification in Population 2 was 97.74%. The negative percent agreement was 100.0%. There were no indeterminate comparator test results for the samples in Population 2. ## c) Clinically Documented Chronic HBV Infection The performance of the VITROS Anti-HBe test was also evaluated with samples from individuals in Population 1 with clinically documented chronic HBV infection. An individual was considered to have clinically documented chronic HBV infection if any one of the following criteria was met: - HBsAg, HBV DNA or HBeAg FDA approved test was positive at least 6 months prior to the current positive HBsAg sample - Documented diagnosis of HBV infection at least 6 months prior to the current positive HBsAg sample - Medical record indicates two positive FDA approved tests for HBsAg, HBV DNA or HBeAg at least 6 months apart Based on the above definitions, 36 individuals from Population 1 were considered to have a chronic HBV infection. An additional 40 chronic HBV samples meeting these criteria were prospectively collected in Moscow, Russia. The subjects were Caucasian and ranged in age from 21 to 77 years. They were 85% male and 15% female. All 40 samples were tested at the testing site in Miami, FL. Thirty-three (82.5%) were positive with the comparator anti-HBe test and 25 (62.5%) were reactive with the VITROS Anti-HBe test. There were no comparator test indeterminate results among the 40 samples. The following table summarizes the positive and negative percent agreement of the VITROS Anti-HBe assay with the comparator anti-HBe test in samples from individuals in the U.S. and Russia with clinically documented chronic HBV infection. Positive and Negative Percent Agreement in Individuals with Chronic HBV Infection (N=76) | Population | Positive Percent Agreement % (N/Total) | 95% Exact Confidence Interval | Negative Percent Agreement % (N/Total) | 95% Exact Confidence Interval | | --- | --- | --- | --- | --- | | Individuals with Chronic HBV Infection from Population 1 (N=36) | 95.65% (22*/23) | 78.05–99.89% | 100.0% (13/13) | 75.29–100.0% | | Individuals with Chronic HBV Infection from Russia (N=40) | 75.76% (25/33) | 57.74–88.91% | 100.0% (7/7) | N/A** | *One VITROS Anti-HBe negative/ comparator indeterminate result was considered VITROS Anti-HBe false negative when calculating positive agreement. ** Confidence intervals calculated on small numbers are not meaningful PMA P100001: FDA Summary of Safety and Effectiveness Data {25} d) Pregnant Women Prospectively collected serum samples from healthy, pregnant women at high risk for exposure to HBV were tested to assess the clinical performance of the assay. Of the 244 women enrolled, there were 229 subjects whose hepatitis marker profiles were consistent with chronically infected, recovered, vaccinated or previously not infected with HBV. Of the 229 prospectively collected samples, 74.7% were obtained in Florida and 25.3% were obtained in Texas. In the population, 18.3% of the pregnant women were in the first trimester, 40.2% were in the second trimester, and 41.5% were in the third trimester of pregnancy. The following table provides a breakdown of the study population. Demographic Profiles of Pregnant Women (N=229) | Collection Site | Florida N (%)* | Texas N (%)* | Total N (%)** | | --- | --- | --- | --- | | Total | 171 (74.7) | 58 (25.3) | 229 (100.0) | | Age (Years) | | | | | 18-20 | 29 (17.0) | 22 (37.9) | 51 (22.3) | | 21-30 | 72 (42.1) | 26 (44.8) | 98 (42.8) | | 31-40 | 63 (36.8) | 9 (15.5) | 72 (31.4) | | 41-50 | 7 (4.1) | 1 (1.7) | 8 (3.5) | | >50 | 0 (0.0) | 0 (0.0) | 0 (0.0) | | Ethnicity | | | | | Caucasian | 4 (2.3) | 9 (15.5) | 13 (5.7) | | African-American | 42 (24.6) | 22 (37.9) | 64 (27.9) | | Hispanic | 107 (62.6) | 23 (39.7) | 130 (56.8) | | Asian | 1 (0.6) | 0 (0.0) | 1 (0.4) | | Indian | 1 (0.6) | 3 (5.2) | 4 (1.7) | | Haitian | 10 (5.8) | 0 (0.0) | 10 (4.4) | | Other | 4 (2.3) | 0 (0.0) | 4 (1.7) | | Unknown | 2 (1.2) | 1 (1.7) | 3 (1.3) | | Trimester | | | | | First | 6 (3.5) | 36 (62.1) | 42 (18.3) | | Second | 78 (45.6) | 14 (24.1) | 92 (40.2) | | Third | 87 (50.9) | 8 (13.8) | 95 (41.5) | * The number (N) of subjects at each site, expressed as a percentage (%) of analyzed subjects at each site. ** The total number (N) of subjects in each category, expressed as a percentage (%) of enrolled subjects (N=229). The following table compares the VITROS Anti-HBe test with the comparator anti-HBe test among the population of pregnant women by trimester. None of the samples had indeterminate results with the comparator anti-HBe test. Comparison of Anti-HBe Test Results in Pregnant Women by Trimester (N=229) | VITROS Anti-HBe Test Result | First Trimester | | | Second Trimester | | | Third Trimester | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Comparator Anti-HBe Test Result | | Total | Comparator Anti-HBe Test Result | | Total | Comparator Anti-HBe Test Result | | Total | | | + | - | | + | - | | + | - | | | Reactive | 0 | 0 | 0 | 1 | 0 | 1 | 0 | 0 | 0 | | Negative | 0 | 42 | 42 | 0 | 91 | 91 | 0 | 95 | 95 | | Total | 0 | 42 | 42 | 1 | 91 | 92 | 0 | 95 | 95 | PMA P100001: FDA Summary of Safety and Effectiveness Data {26} Positive and negative percent agreement between the VITROS Anti-HBe test and the comparator anti-HBe test were calculated for the pregnant women in this study group. The following table summarizes these calculations and provides the 95% exact confidence intervals, where appropriate. Positive and Negative Percent Agreement by HBV Disease Classification among Pregnant Women (N=229) | HBV Disease Classification | Positive Percent Agreement % (N/Total) | 95% Exact Confidence Interval | Negative Percent Agreement % (N/Total) | 95% Exact Confidence Interval | | --- | --- | --- | --- | --- | | Chronic | 100.0% (1/1) | N/A* | 100.0% (2/2) | N/A* | | Recovered | N/A (0/0)** | N/A | 100.0% (11/11) | 71.51–100.0% | | HBV Vaccine Response | N/A (0/0)** | N/A | 100.0% (27/27) | 87.23–100.0% | | Not Previously Infected with HBV | N/A (0/0)** | N/A | 100.0% (188/188) | 98.06–100.0% | * Confidence intervals calculated on small numbers are not meaningful. ** There were no subjects with this HBV disease classification whose comparator test results fell within this category. e) Clinical Performance in Pediatric Subjects Performance of the VITROS Anti-HBe assay in pediatric serum was determined using prospective samples from a population of pediatric subjects in Florida (N=165). The group was 47.9% male and 52.1% female, and the subjects' ages ranged from 2 through 21 years. The following table compares the VITROS Anti-HBe results with the comparator anti-HBe results for the pediatric subjects. There were no comparator indeterminate results among the pediatric subjects. Comparison of Anti-HBe Test Results in Pediatric Subjects (N=165) | VITROS Anti-HBe Test Result | Comparator Anti-HBe Test Result | | | Total N | | --- | --- | --- | --- | --- | | | Positive N | Negative N | Indeterminate N | | | Reactive | 0 | 0 | 0 | 0 | | Negative | 0 | 165 | 0 | 165 | | Total | 0 | 165 | 0 | 165 | The following table summarizes the percent agreement between the VITROS Anti-HBe test and the comparator anti-HBe test for the pediatric population. The table provides the 95% exact confidence intervals. Positive and Negative Percent Agreement in Pediatric Subjects (N=165) | Subjects | Positive Percent Agreement % (N/Total) | 95% Exact Confidence Intervals | Negative Percent Agreement % (N/Total) | 95% Exact Confidence Intervals | | --- | --- | --- | --- | --- | | Pediatric | N/A (0/0) | N/A | 100.0% (165/165) | 97.79–100.0% | PMA P100001: FDA Summary of Safety and Effectiveness Data {27} There was 100% concordance between the VITROS Anti-HBe test and the FDA approved comparator anti-HBe test. None of the 165 samples was reactive with either the VITROS Anti-HBe test or the comparator anti-HBe test. There were no indeterminate comparator test results for the pediatric population. The VITROS Anti-HBe assay performance was also evaluated using spiked anti-HBe reactive pediatric samples. Thirty (30) individual pediatric samples, non-reactive for anti-HBe, were spiked with an anti-HBe positive patient sample to a target level of 2.00–4.00 C/S and compared to matched spikes of an adult pool derived from a base matrix (pooled, defibrinated adult plasma, clarified, dialyzed and filtered). Ten (33%) samples were from subjects 2 to 11 years old and 20 (67%) were from subjects 12 to 21 years old. Each sample was run in the VITROS Anti-HBe test in duplicate. Mean results from the 30 spiked sample pairs were used to calculate the percent difference between the pediatric and the adult pool spike. Twenty-seven of the 30 spiked pediatric samples gave reactivity lower than the spiked adult pool (derived from base matrix) ranging from 0.3% to 35.8% lower (average difference was 10.1% lower). Three of the 30 spiked pediatric samples gave reactivity higher than the adult pool, ranging from 0.7% to 8.1% higher (average difference was 3.5% higher). ## XI. PANEL MEETING RECOMMENDATION AND FDA'S POST-PANEL ACTION In accordance with the provisions of section 515(c)(2) of the act as amended by the Safe Medical Devices Act of 1990, this PMA was not referred to the FDA 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. ## XII. CONCLUSIONS DRAWN FROM PRECLINICAL AND CLINICAL STUDIES ### A. Safety Conclusions The adverse effects of the device are based on data collected in a clinical study conducted to support PMA approval as described above. As a diagnostic test, the VITROS Anti-HBe assay involves removal of blood from an individual for testing purposes. The test, therefore, presents no more safety hazard to an individual being tested than other tests where blood is removed. There were no adverse effects of the device reported while the study was conducted. ### B. Effectiveness Conclusions - The sensitivity and specificity of the VITROS Anti-HBe assay was shown to be comparable with the current commercially available FDA approved anti- PMA P100001: FDA Summary of Safety and Effectiveness Data {28} HBe assay in patients who are chronically infected with hepatitis or who have recovered from hepatitis B infection. - The comparison of the performance of the VITROS Anti-HBe test in patients with chronic HBV infection among the study subjects, demonstrated a &gt; 95% positive and negative percent agreement with the FDA approved comparator anti-HBe test. - The comparison of the performance of the VITROS Anti-HBe test in those patients among the study subjects who recovered from HBV infection, demonstrated a &gt; 99% positive and negative percent agreement with the FDA approved comparator anti-HBe test. - The comparison of the performance of the VITROS Anti-HBe test in patients who were never previously infected or have been vaccinated against HBV demonstrated a &gt; 99% negative percent agreement with the FDA approved comparator anti-HBe test. - The performance of the VITROS Anti-HBe test was shown to be acceptable in pregnant women. - The performance of the VITROS Anti-HBe test was shown to be acceptable in serum from pediatric patients (2 to 21 years old). - Studies have shown that the VITROS Anti-HBe test has no significant cross-reactivity with the potentially cross-reacting clinical subgroups. - Seroconversion sensitivity of the VITROS Anti-HBe assay has been shown to be acceptable by testing six commercial seroconversion panels. - The stability of the VITROS Anti-HBe Reagent Pack and Calibrator has been demonstrated for a period of up to 40 weeks when stored at 2-8 °C. - The stability of the VITROS Anti-HBe Reagent Pack and Calibrator, when stored on-board of the analyzer, has been demonstrated for a period of up to 12 weeks. - The calibration interval is stable for 28 days when using the same lot of reagents. - The stability of the VITROS Anti-HBe Controls has been demonstrated for a period of 52 weeks when stored at 2-8 °C. Once reconstituted, the Controls are stable for 5 days at 2-8 °C. - The demonstrated precision of the VITROS Anti-HBe assay is within the expected range of this type of device. The results from both the non-clinical and clinical studies indicate that the VITROS Anti-HBe assay is safe and effective for the in vitro qualitative detection of antibodies to the hepatitis B e antigen (Anti-HBe) in human adult and pediatric serum. ## C. Overall Conclusions The data in this application support a reasonable assurance of safety and effectiveness of this device when used in accordance with the indications for use. The submitted clinical studies have shown that the VITROS Anti-HBe assay, when compared to FDA approved comparator, has a similar ability to detect the presence of anti-HBe antibodies in serum specimens from individuals with chronic hepatitis B, or those PMA P100001: FDA Summary of Safety and Effectiveness Data {29} recovered from HBV infection. The rate of false positivity and false negativity are within acceptable limits compared to the comparator assay. It has been shown that the device has no demonstrable cross-reactivity with antibodies found in patients with potentially cross-reacting medical conditions. Therefore, this device should benefit the physician in providing additional information about a patient's progression to seroconversion which is important in the management of HBV infection. ## XIII. CDRH DECISION FDA issued an approval order on July 20, 2011. The final conditions of approval are cited in the approval order. The applicant’s manufacturing facilities were inspected and found to be in compliance with the devices Quality System (QS) regulation (21 CFR 820) on June 24, 2011. ## 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 P100001: FDA Summary of Safety and Effectiveness Data page 30 34
Innolitics

Panel 1

/
Sort by
Ready

Predicate graph will load when search results are available.

Embedding visualization will load when search results are available.

PDF viewer will load when search results are available.

Loading panels...

Select an item from Submissions

Click any panel, subpart, regulation, product code, or device to see details here.

Section Matches

Results will appear here.

Product Code Matches

Results will appear here.

Special Control Matches

Results will appear here.

Loading collections...