Immunoassay for the in vitro qualitative detection of total antibodies (IgG and IgM) to hepatitis A virus (HAV) in human pediatric (ages 2 through 21 years) and adult serum and plasma (Li-heparin, potassium EDTA, Na-hebarin). The assay, in conjunction with other serological and clinical information, is indicated as an aid in the clinical laboratory diagnosis of acute or past hepatitis A virus infection in persons with signs or symptoms of hepatitis and in persons at increased risk for hepatitis A infection, or as an aid to identify HAV susceptible individuals and to determine the presence of an antibody response to HAV in vaccine recipients.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on the cobas e immunoassay analyzers.

Assay performance characteristics have not been established for immunocompromised or immunosuppressed patients. This assay has not been FDA cleared or approved for the screening of blood or plasma donors.

Device Description

Elecsys Anti-HAV II is a second generation assay by Roche Diagnostics for the in vitro qualitative detection of total antibodies (IgG and IgM) to the hepatitis A virus (HAV) in human pediatric (ages 2 through 21 years) and adult serum and plasma. It is intended for use on the cobas e 601 immunoassay analyzer. The cobas e family of analyzers employs the electrochemiluminescence "ECLIA" technology. The assay is an 18-minute assay utilizing a competition principle.

Results are determined automatically by the software by comparing the electrochemiluminescence signal obtained from the reaction product of the sample with the signal of the cutoff value previously obtained by calibration.

The reagent rackpack working solutions include:

M: Streptavidin-coated microparticles .
R1: HAV Ag (cell culture) .
R2: Biotinylated monoclonal anti-HAV antibody, monoclonal Anti-HAV antibody . labeled with ruthenium complex
AHAV 2 Cal1: Negative Calibrator 1 (human serum) .
AHAV 2 Cal2: Positive Calibrator 2 (anti-HAV (human), approximately 60 IU/L in . human serum)

PreciControl Anti-HAV II is a ready-for-use control serum based on human serum both in the negative and positive concentration range. The controls are used for monitoring the performance of the Elecsys Anti-HAV II immunoassay. PreciControl Anti-HAV II is sold separately from the Elecsys Anti-HAV II immunoassay reagent.

AI/ML Overview

The Elecsys Anti-HAV II is an immunoassay for the in vitro qualitative detection of total antibodies (IgG and IgM) to hepatitis A virus (HAV) in human pediatric (ages 2 through 21 years) and adult serum and plasma. Its purpose is to aid in the clinical laboratory diagnosis of acute or past hepatitis A virus infection in persons with signs or symptoms of hepatitis and in persons at increased risk for hepatitis A infection, or as an aid to identify HAV susceptible individuals and to determine the presence of an antibody response to HAV in vaccine recipients.

Here's an analysis of the acceptance criteria and the studies performed:

1. Table of Acceptance Criteria and Reported Device Performance:

Study Category	Acceptance Criteria (General)	Elecsys Anti-HAV II Performance
Non-Clinical Performance
Precision (Repeatability, Within-Lab)	CV values within acceptable limits for various COI levels and controls.	Repeatability CV: 0.9-3.0% for human sera, 1.1-1.5% for controls. Within-Laboratory Precision CV: 1.9-3.3% for human sera, 1.8-2.7% for controls. All results met pre-defined acceptance criteria.
Analytical Sensitivity (Cut-off)	Cut-off sensitivity established/validated to correspond to ≤ 25.4 IU/L.	The cut-off of COI = 1.0 corresponds to ≤ 25.4 IU/L. Individual reagent lots showed cut-off sensitivities of 24.6 IU/L, 25.4 IU/L, and 20.1 IU/L.
HAMA Interference	No HAMA interference within pre-defined acceptance criteria.	No HAMA interference was found within the predefined acceptance criteria using 11 human serum samples.
Endogenous Interferences	No interference from tested substances up to specified levels.	No interference seen up to: Intralipid (2000 mg/dL), Bilirubin (66 mg/dL), Hemoglobin (1000 mg/dL), Rheumatoid Factor (1400 IU/mL), human Serum albumin (7.00 g/dL), human IgG (7.00 g/dL), human IgM (1.00 g/dL), human IgA (1.60 g/dL). All results met pre-defined acceptance criteria.
Biotin Interference	Negative bias in COI values for biotin concentrations up to 100 ng/mL ≤11%.	Negative specimens with biotin concentrations up to 100 ng/mL demonstrated ≤11% negative bias in COI values. Concentrations > 100 ng/mL lead to higher negative bias and can result in false positives.
Analytical Specificity/Cross-Reactivity	No cross-reactivity with other infectious agents.	No cross-reactivity found with samples containing antibodies to various infectious diseases (e.g., acute Hepatitis B/C, HIV, EBV, CMV, HSV, Toxoplasma Gondii, Treponema Pallidum, Mumps/Rubeola, Rubella, Parvovirus B19, ANA Autoimmune).
Exogenous Interferences (Drugs)	No interference from tested drug substances at specified concentrations.	All results met pre-defined acceptance criteria, demonstrating no interference from 18 commonly used pharmaceutical compounds at tested concentrations (at least five times maximum daily doses).
Sample Matrix Comparison	All anti-coagulants (K2-EDTA, K3-EDTA, Na-Heparin, Li-Heparin, Na-Citrate) are acceptable.	Specifications met for all tested anti-coagulants (K2-EDTA, K3-EDTA, Na-Heparin, Li-Heparin, Na-Citrate), demonstrating they are acceptable for use with Elecsys Anti-HAV II. (≥ 60 serum/plasma pairs tested for each type).
Analytical Method Comparison	Equivalency between candidate and predicate devices, meeting pre-defined acceptance criteria.	Positive percent agreement: 100% (107/107). Negative percent agreement: 100% (98/98). All results met pre-defined acceptance criteria, demonstrating equivalency with the predicate device (Elecsys Anti-HAV).
Reagent Stability	Reagent stability after first opening (8 weeks at 2-8°C). On-board reagent stability (8 weeks).	All pre-defined acceptance criteria were met for: 8 weeks stability after first opening at 2-8°C. 8 weeks on-board stability on the cobas e 601 immunoassay analyzer (new calibration recommended every 7 days).
Calibration Stability	Lot calibration stability (4 weeks). On-board calibration stability (1 week).	All pre-defined acceptance criteria were met for: Lot calibration stability (renewed calibration recommended after 4 weeks). On-board calibration stability (can be used for up to one week when using the same lot of reagents stored on the analyzer).
Sample Stability	Stability under various storage conditions: 2-8°C (14 days), RT (6 days), -15 to -25°C (3 months), 5 freeze/thaw cycles.	All pre-defined acceptance criteria were met, demonstrating stability for: 14 days at 2-8°C. 6 days at 15-25°C. 3 months at -15 to -25°C. 5 freeze/thaw cycles.
Clinical Performance (External Testing)
Reproducibility	Repeatability, within-laboratory, and reproducibility SD and CV values within acceptable limits.	Repeatability CV: 1.1-1.7%. Within-Laboratory (between run, between day, between site) components contribute to overall reproducibility. Overall Reproducibility CV: 2.2-4.0%. All results met pre-defined acceptance criteria.
Method Comparison vs. Predicate (Overall)	Lower bound of the 95% CI for agreement rates (PPA and NPA) ≥ 90%.	Overall PPA: 99.80% (501/502), 95% CI (98.90, 99.99). Overall NPA: 95.21% (437/459), 95% CI (92.83, 96.97). All overall percentages met the expected performance.
Pre- and Post-HAV Vaccination	No discrepant results between Elecsys Anti-HAV II and predicate assay.	No discrepant results observed in 49 subjects evaluated both pre- and post-HAV vaccination.

2. Sample Size Used for the Test Set and Data Provenance:

Precision (Repeatability and Within-Laboratory Precision): 5 human serum sample pools and 2 PreciControl materials, tested in 2 aliquots per run, 2 runs per day for 21 days (total 84 measurements per sample/control).
Analytical Sensitivity (Cut-off): 10 serially diluted samples of the 2nd International Standard for Anti-Hepatitis A, Immunoglobulin, Human, NIBSC code: 97/646. Tested fourfold with three different reagent and calibrator lots.
HAMA Interference: 11 human serum samples, double positive for HAMA and anti-HAV antibodies.
Endogenous Interferences (Hemoglobin/Bilirubin/Lipemia, Rheumatoid Factor, Serum Albumin/IgG/IgA/IgM): Four human serum samples for each interfering substance, tested in accordance with CLSI EP07-A2.
Biotin Interference: Five human serum samples.
Analytical Specificity/Cross-Reactivity: 12 sample pools, each containing 10 samples (total 120 samples) with antibodies to various infectious agents.
Exogenous Interferences (Drugs): Four human serum samples (native human serum pools) for each of 18 pharmaceutical compounds.
Sample Matrix Comparison: At least 60 serum/plasma pairs for each anticoagulant type (K2-EDTA, K3-EDTA, Na-Heparin, Li-Heparin, Na-Citrate).
Analytical Method Comparison to Predicate: A total of 215 modified serum samples (≥100 negative and ≥100 positive samples).
Reagent Stability:
- After First Opening: 2 control samples and 7 samples, tested in duplicate.
- On-Board Reagent Stability: 2 control samples (singleton) and 7 samples (duplicate).
Calibration Stability:
- Lot Calibration Stability: 2 control samples (singleton) and 7 human serum samples (duplicate).
- On-Board Calibration Stability: 2 control samples (singleton) and 7 human serum samples (duplicate).
Sample Stability:
- 2-8°C, Room Temperature, -15 to -25°C, Freeze/Thaw Cycles: 6 human serum, K2-EDTA, Li-Heparin, and Na-Citrate plasma samples, and 7 K3-EDTA and Na-Heparin plasma samples.
Reproducibility (External Testing): Four human serum pools and two PreciControl materials. Tested in 3 replicates per run, 2 runs per day for 5 days (total 90 measurements per sample/control per site).
Method Comparison Versus Predicate (Clinical): 961 specimens from various cohorts.
- Routine HAV testing: 91 PPA, 109 NPA
- Hospitalized: 56 PPA, 144 NPA
- Increased risk for hepatitis: 119 PPA, 87 NPA
- Symptomatic: 129 PPA, 91 NPA
- Characterized acute HAV: 65 PPA, 10 NPA
- Pediatric: 42 PPA, 18 NPA
Pre- and Post-HAV Vaccination: Specimens from 49 subjects.
Prevalence Study: 400 evaluable subjects from a "high prevalence" region (Western US) and 400 evaluable subjects from a "low prevalence" region (Eastern US).

Data Provenance for Clinical Studies:
The method comparison and prevalence studies were conducted using specimens that are likely from the United States, as they reference "US- and non-US-obtained specimens" and "Western United States" and "Eastern United States" for the prevalence study. These studies appear to be prospective in nature, as they involve specimen collection for specific study purposes (e.g., cohorts for method comparison, pre- and post-vaccination, and prevalence studies with subjects recruited from specific regions).

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications:

The document does not explicitly state the number or qualifications of "experts" used to establish the ground truth in the traditional sense of consensus reading or clinical adjudication by medical specialists.

For the method comparison study, the "ground truth" was established by the predicate device, Elecsys Anti-HAV assay (FDA-cleared). This means the predicate device's results were accepted as the reference against which the new device's performance was measured.
For the Pre- and Post-HAV Vaccination study, the "ground truth" was established by the predicate assay (Elecsys Anti-HAV).
For the Analytical Specificity/Cross-Reactivity study, samples were defined as "Anti-HAV negative samples containing potential cross-reacting antibodies to other infectious diseases," implying these diagnoses were previously established, likely through standard diagnostic tests.
For the Prevalence study, the "ground truth" for HAV antibody status was determined by the Elecsys Anti-HAV II assay itself, as the study evaluated its performance in determining prevalence in different populations rather than comparing it to an external gold standard for individual cases.

4. Adjudication Method for the Test Set:

None explicitly mentioned in the typical sense of expert review for ambiguous cases.
For the Method Comparison study, discordant results were counted against the Elecsys Anti-HAV II when calculating agreement. Specifically, specimens with results in the borderline range of the predicate device (18.0 ≤ IU/L < 22.0) were counted as discordant with the Elecsys Anti-HAV II assay. This indicates a pre-defined rule for handling discrepancies relative to the predicate, rather than an expert adjudication process.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done:

No, an MRMC comparative effectiveness study was not done. This device is an in vitro diagnostic (IVD) immunoassay, not an imaging device that would typically involve multiple human readers interpreting images. The studies performed compare the new device's performance to a predicate device or pre-defined analytical acceptance limits, not to human reader performance with or without AI assistance.

6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) was done:

Yes, the studies are inherently standalone. The Elecsys Anti-HAV II is an automated immunoassay designed for use on cobas e immunoassay analyzers. All performance evaluations described (precision, sensitivity, specificity, interference, stability, and method comparison) are based solely on the algorithm and analytical performance of the device itself, with no human interpretation of individual assay results affecting the determination of the final qualitative output (Reactive/Non-Reactive). The human role is limited to performing the test and reviewing the results generated by the analyzer.

7. The Type of Ground Truth Used:

The primary ground truth for the comparative studies was the predicate device (Elecsys Anti-HAV assay), which is an FDA-cleared immunoassay. This indicates that the predicate device's results were considered the reference standard for classification of samples as Reactive, Borderline, or Non-Reactive for HAV antibodies.
For other studies (e.g., cross-reactivity), samples were selected based on established diagnoses of other infectious agents.
For precision and stability studies, the results are evaluated against internal consistency or expected recovery values rather than an external ground truth for individual samples.
For the analytical sensitivity, the ground truth was the 2nd International Standard for Anti-Hepatitis A, Immunoglobulin, Human (NIBSC code: 97/646).

8. The Sample Size for the Training Set:

The document does not provide information on a specific "training set" in the context of machine learning or AI algorithm development. This is an immunoassay (IVD), not an AI/ML-based device in the typical sense. Data described are for analytical and clinical validation.

9. How the Ground Truth for the Training Set Was Established:

As a "training set" is not described for this immunoassay, this question is not applicable. The device's cut-off and performance characteristics are established through internal studies (e.g., "internal studies" for cut-off determination) and validated through the non-clinical and clinical performance evaluations described.

Summary

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the square are the words "U.S. FOOD & DRUG" in blue, with the word "ADMINISTRATION" below it.

August 13, 2019

Roche Diagnostics Jamie Ferguson Regulatory Affairs Principal 9115 Hague Road Indianapolis, Indiana 46250

Re: K190428

Trade/Device Name: Elecsys Anti-HAV II Regulation Number: 21 CFR 866.3310 Regulation Name: Hepatitis A Virus (HAV) Serological Assays Regulatory Class: Class II Product Code: LOL, QCH Dated: February 20, 2019 Received: February 22, 2019

Dear Jamie Ferguson:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's

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requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

for

Maria Garcia, Ph.D. Chief Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K190428

Device Name Elecsys Anti-HAV II

Indications for Use (Describe)

The electrochemiluminescence immunoassay "ECLIA" is intended for use on the cobas e immunoassay analyzers.

Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)

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Elecsys Anti-HAV II 510(k) Summary

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.

In accordance with 21 CFR 807.87, Roche Diagnostics hereby submits official notification as required by Section 510(k) of the Federal Food, Drug and Cosmetics Act of our intention to market the device described in this Premarket Notification 510(k).

The purpose of this Traditional 510(k) Premarket Notification is to obtain FDA review and clearance for the Elecsys Anti-HAV II.

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Submitter Name	Roche Diagnostics
Address	9115 Hague RoadP.O. Box 50416Indianapolis, IN 46250-0457
Contact	Jamie FergusonPhone: (317) 521-4213Fax: (317) 521-2324Email: Jamie.ferguson@roche.com
	Tammy DeanPhone: (317) 521-3978Fax: (317) 521-2324Email: tammy.dean@roche.com
Date Prepared	February 20, 2019
Proprietary Name	Elecsys Anti-HAV IIPreciControl Anti-HAV II
Common Name	Anti-HAV IIPreciControl Anti-HAV II
Classification Name	1. Hepatitis A virus (HAV) Serological Assays2. Assayed Quality Control Material for Clinical Microbiology Assays
Product Codes,Regulation Numbers	1. LOL, 21 CFR §866.33102. QCH, 21 CFR §866.3920, Quality Control
Predicate Devices	Elecsys Anti-HAV (K100903)
Establishment Registration	For the Elecsys Anti-HAV II, the establishment registration number for RocheDiagnostics GmbH in Mannheim, Germany is 9610126, and for Penzberg,Germany, 9610529. The establishment registration number for RocheDiagnostics in the United States is 1823260.

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DEVICE DESCRIPTION 1.

The reagent rackpack working solutions include:

M: Streptavidin-coated microparticles .
R1: HAV Ag (cell culture) .
R2: Biotinylated monoclonal anti-HAV antibody, monoclonal Anti-HAV antibody . labeled with ruthenium complex
AHAV 2 Cal1: Negative Calibrator 1 (human serum) .
AHAV 2 Cal2: Positive Calibrator 2 (anti-HAV (human), approximately 60 IU/L in . human serum)

2. INTENDED USE

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serological and clinical information, is indicated as an aid in the clinical laboratory diagnosis of acute or past hepatitis A virus infection in persons with signs or symptoms of hepatitis and in persons at increased risk for hepatitis A infection, or as an aid to identify HAV susceptible individuals and to determine the presence of an antibody response to HAV in vaccine recipients.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on the cobas e immunoassay analyzers.

3. TECHNOLOGICAL CHARACTERISTICS

Elecsys Anti-HAV II utilizes electrochemiluminescence "ECLIA" technology for the qualitative detection of total antibodies (IgG and IgM) to the hepatitis A virus (HAV) in human pediatric (ages 2 through 21 years) and adult serum and plasma on the cobas e 601 immunoassay analyzer.

The following tables compare the Elecsys Anti-HAV II with its predicate device, Elecsys Anti-HAV (K100903).

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Assay Comparison Table 1:

Feature	Elecsys Anti-HAV (K100903)	Elecsys Anti-HAV II
Intended Use/Indications forUse	Immunoassay for the in vitro qualitativedetection of total antibodies (IgM andIgG) to hepatitis A virus in human serumand plasma (K2-EDTA). The assay isintended for use as an aid in thelaboratory diagnosis of past oracute/recent hepatitis A infection. Assayresults, in conjunction with otherlaboratory results and clinicalinformation, may be used to providepresumptive evidence of infection withhepatitis A virus in persons with signs orsymptoms of hepatitis and in persons atrisk for hepatitis A infection, or used as anaid to determine the presence of antibodyresponse to HAV in vaccine recipients.The electrochemiluminescenceimmunoassay "ECLIA" is intended foruse on Elecsys and cobas e immunoassayanalyzers.This assay is not intended for screeningblood or solid or soft tissue donors. Assayperformance characteristics have not beenestablished for immunocompromised orimmunosuppressed patients. The users areresponsible for establishing their ownassay performance characteristics in thesepopulations.	Immunoassay for the in vitro qualitativedetection of total antibodies (IgG andIgM) to hepatitis A virus (HAV) inhuman pediatric (ages 2 through 21years) and adult serum and plasma (Li-Heparin, potassium EDTA, Na-Citrate,Na-Heparin). The assay, in conjunctionwith other serological and clinicalinformation, is indicated as an aid in theclinical laboratory diagnosis of acute orpast hepatitis A virus infection inpersons with signs or symptoms ofhepatitis and in persons at increased riskfor hepatitis A infection, or as an aid toidentify HAV susceptible individualsand to determine the presence of anantibody response to HAV in vaccinerecipients.The electrochemiluminescenceimmunoassay "ECLIA" is intended foruse on cobas e immunoassay analyzers.Assay performance characteristics havenot been established forimmunocompromised orimmunosuppressed patients. This assayhas not been FDA cleared or approvedfor the screening of blood or plasmadonors.
Assay Method	Competition principle binding protein	Same
Detection Method	Electrochemiluminescence	Same
Applications/Test Time	18 minutes	Same
Instrument Platform	Elecsys 2010, cobas e 411, cobas e 601,cobas e 602, and MODULARANALYTICS E170	cobas e 601
Sample volume	50 µL	20 µL
Sample Type/Matrix	Human serum, plasma	Same
Sample Anticoagulants	K2-EDTA	Li-Heparin, potassium EDTA, Na-Citrate, Na-Heparin
Calibrator	Anti-HAV Cal1 and Cal2 (packed in kit)	Anti-HAV II Cal1 and Cal2 (packed inkit)
Calibration Method	2-point calibration	Same
Feature	Elecsys Anti-HAV (K100903)	Elecsys Anti-HAV II
Calibration Interval	Calibration must be performed once perreagent lot using fresh reagent (i.e. notmore than 24 hours since the reagent kitwas registered on the analyzer). Renewedcalibration is recommended as follows:• after 1 month (28 days) when usingthe same reagent lot• after 7 days (when using the samereagent kit on the analyzer)• as required: e.g. quality controlfindings outside the defined limits	Same
Controls	PreciControl Anti-HAV	PreciControl Anti-HAV II
Traceability/Standardization	Second International Standard for AntiHepatitis A, Immunoglobulin, Human,NIBSC code: 97/646	Same
Reagent Stability	Store at 2-8°C.Do not freeze.Store the Elecsys reagent kit upright inorder to ensure complete availability ofthe microparticles during automaticmixing prior to use.Unopened at 2-8°C, up to statedexpiration date.After opening at 2-8°C, 8 weeks.On the analyzers at 20-25°C, 7 days or 4weeks when stored alternative in therefrigerator and on the analyzer, with thetotal time onboard on the analyzer notexceeding 40 hours.	Store at 2-8°C.Do not freeze.Store the Elecsys reagent kit upright inorder to ensure complete availability ofthe microparticles during automaticmixing prior to use.Unopened at 2-8°C, up to statedexpiration date.After opening at 2-8°C, 8 weeks.On the analyzers at 20-25°C, 8 weeks.
Feature	Elecsys Anti-HAV (K100903)	Elecsys Anti-HAV II
Limitations	Bilirubin < 855 µmol/L or < 50 mg/dLHemolysis < 0.623 mmol/L or < 1.0 g/dLLipemia < 1500 mg/dLBiotin < 205 nmol/L or < 50 ng/mLCriterion: Recovery within +/- 20% ofinitial valueSamples should not be taken from patientsreceiving therapy with high biotin doses(i.e. > 5 mg/day) until at least 8 hoursfollowing the last biotin administration.In vitro tests were performed on 18commonly used pharmaceuticals(Acetylcystein, Ampicillin, Ascorbic acid,Ca Dobesilate, Cyclosporine, Cefoxitin,Heparin, Intralipid, Levodopa,Methyldopa, Metronidazole,Phenylbutazone, Tetracycline,Acetylsalicylic Acid, Rifampicin,Acetaminophen, Ibuprofen andTheophylline). No interference with theassay was found.In rare cases, interference due toextremely high titers of antibodies toanalyte specific antibodies, streptavidin orruthenium can occur. These effects areminimized by suitable test design.	Bilirubin < 1129 µmol/L or ≤ 66 mg/dLHemoglobin ≤ 0.621 mmol/L or ≤ 1000mg/dLIntralipid ≤ 2000 mg/dLRheumatoid Factors ≤ 1400 IU/mLIgG ≤ 7.0 g/dLIgA ≤ 1.6 g/dLIgM ≤ 1.0 g/dLSerum Albumin ≤ 7 g/dLCriterion: > 1.0 COI +/- 20% recovery,≤ 1.0 COI +/- 0.20 recoveryNegative specimens with biotinconcentrations up to 100 ng/mLdemonstrated ≤ 11 % negative bias inCOI values. Biotin concentrationsgreater than 100 ng/mL lead to highernegative bias and in consequence canlead to false positive Elecsys Anti-HAVII results. Some studies have shown thatserum concentrations of biotin can reachup to 355 ng/mL within the first hourafter ingestion for subjects consumingsupplements of 20 mg biotin per day andup to 1160 ng/mL in plasma for subjectsconsuming a single dose of 300 mgbiotin.In vitro tests were performed on 18commonly used pharmaceuticals(Acetylcystein, Ampicillin-Na, Ascorbicacid, Ca Dobesilate, Cyclosporine,Cefoxitin, Doxycycline, Heparin,Levodopa, Methyldopa +1.5,Metronidazole, Phenylbutazone,Tetracycline, Acetylsalicylic Acid,Rifampicin, Acetaminophen, Ibuprofenand Theophylline). No interference withthe assay was found.In rare cases, interference due toextremely high titers of antibodies toanalyte specific antibodies, streptavidinor ruthenium can occur. These effects areminimized by suitable test design.

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NON-CLINICAL PERFORMANCE EVALUATION 4.

Non-clinical performance evaluation for Elecsys Anti-HAV II is briefly summarized below.

Precision 4.1.

4.1.1. Repeatability and Within-Laboratory Precision

Precision measurements were conducted to evaluate repeatability (within-run precision) and within-laboratory precision (intermediate precision) in a study based on the protocol of CLSI EP05-A3. Precision was evaluated on a single cobas e 601 immunoassay analyzer. One Elecsys Anti-HAV II reagent lot was evaluated. The protocol consisted of testing 2 aliquots each of two levels of control and 5 human sera per run, 2 runs per day for 21 days. Calibration was performed on day 1 and on day 17. Serum samples were human serum sample pools.

		Repeatability		Within-Laboratory(Intermediate) Precision
Sample	Mean (COI)	SDa)(COI)	CV (%)	SD(COI)	CV (%)	n
Human serum 1	1.42	0.016	1.1	0.028	2.0	84
Human serum 2	1.15	0.012	1.1	0.022	1.9	84
Human serum 3	0.955	0.009	0.9	0.022	2.3	84
Human serum 4	0.665	0.009	1.3	0.020	2.9	84
Human serum 5	0.006	0.0002	3.0	0.0002	3.3	84
PCb) Anti-HAV II 1	1.30	0.015	1.1	0.024	1.8	84
PC Anti-HAV II 2	0.339	0.005	1.5	0.009	2.7	84

Table 2: Repeatability and Within-Laboratory Precision Results

SD = Standard Deviation a)

PC = PreciControl b)

4.2. Analytical Sensitivity

4.2.1. Determination of Cut-off Sensitivity

The cut-off of the Elecsys Anti-HAV II assay was established with internal studies, and the validation of the cut-off was performed by external clinical studies. In order to determine the cut-off sensitivity, the 2nd International Standard for Anti-Hepatitis A, Immunoglobulin, Human,

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NIBSC code: 97/646 was serially diluted to 10 samples and tested fourfold with three different reagent and calibrator lots. The cut-off of COI = 1.0 corresponds to ≤ 25.4 IU/L.

Dilution-factor/Sample	Target value[IU/L]	Measuring cell 1 [COI]		Measuring cell 2 [COI]		[COI]Mean
		Run 1	Run 2	Run 1	Run 2
Ref-Std_01	0	1.31	1.28	1.31	1.30	1.30
Ref-Std_02	10.2	1.16	1.17	1.17	1.16	1.17
Ref-Std_03	20.3	1.05	1.03	1.06	1.04	1.05
Ref-Std_04	30.5	0.921	0.923	0.923	0.946	0.928
Ref-Std_05	59.9	0.628	0.626	0.637	0.646	0.634
Ref-Std_06	99.7	0.365	0.361	0.365	0.369	0.365
Ref-Std_07	200	0.098	0.098	0.104	0.102	0.100
Ref-Std_08	300	0.035	0.034	0.036	0.035	0.035
Ref-Std_09	400	0.017	0.017	0.017	0.017	0.017
Ref-Std_10	500	0.011	0.011	0.019	0.011	0.013
Cut-off sensitivity		24.6 IU/L

Table 3: Lot MP01

Table 4: Lot PoQ

Dilution-factor/Sample	Target value[IU/L]	Measuring cell 1 [COI]		Measuring cell 2 [COI]		[COI]Mean
		Run 1	Run 2	Run 1	Run 2
Ref-Std_01	0	1.28	1.28	1.27	1.28	1.28
Ref-Std_02	10.2	1.14	1.15	1.17	1.17	1.16
Ref-Std_03	20.3	1.05	1.06	1.04	1.04	1.05
Ref-Std_04	30.5	0.941	0.944	0.934	0.942	0.940
Ref-Std_05	59.9	0.663	0.663	0.674	0.665	0.667
Ref-Std_06	99.7	0.389	0.399	0.399	0.401	0.397
Ref-Std_07	200	0.113	0.112	0.115	0.115	0.114
Ref-Std_08	300	0.039	0.038	0.040	0.040	0.040
Ref-Std_09	400	0.018	0.018	0.018	0.019	0.018
Ref-Std_10	500	0.011	0.011	0.011	0.011	0.011
Cut-off sensitivity				25.4 IU/L

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Dilution-factor/Sample	Target value[IU/L]	Measuring cell 1 [COI]		Measuring cell 2 [COI]		[COI]Mean
		Run 1	Run 2	Run 1	Run 2
Ref-Std_01	0	1.25	1.25	1.26	1.27	1.26
Ref-Std_02	10.2	1.11	1.10	1.13	1.15	1.12
Ref-Std_03	20.3	0.976	0.975	0.982	0.984	0.980
Ref-Std_04	30.5	0.867	0.859	0.873	0.872	0.868
Ref-Std_05	59.9	0.551	0.548	0.563	0.559	0.555
Ref-Std_06	99.7	0.269	0.274	0.276	0.276	0.274
Ref-Std_07	200	0.048	0.048	0.049	0.048	0.048
Ref-Std_08	300	0.016	0.016	0.016	0.016	0.016
Ref-Std_09	400	0.010	0.010	0.010	0.010	0.010
Ref-Std_10	500	0.009	0.009	0.009	0.009	0.009
Cut-off sensitivity		20.1 IU/L

Table 5: Lot P02

4.3. Human Anti-Mouse Antibodies (HAMA)

The effect on detection of analyte in the presence of HAMA using the Elecsys Anti-HAV II was determined on the cobas e 601 immunoassay analyzer using 11 human serum samples. Only samples that are double positive for HAMA and anti-HAV antibodies were used, since HAMA interference would lead to false negative results in the competitive assay format. No HAMA interference was found within the predefined acceptance criteria.

Endogenous Interferences 4.4.

Hemoglobin/Bilirubin/Lipemia 4.4.1.

The purpose of this study was to evaluate endogenous substances for potential interference with the parameters measured on the Elecsys Anti-HAV II on the cobas e 601 immunoassay analyzer. For each interfering substance, four human serum samples were tested in accordance with CLSI EP07-A2.

4.4.2. Rheumatoid Factor Interference

The recovery of analyte values in the presence of endogenous interfering substances was determined with the Elecsys Anti-HAV II on the cobas e 601 immunoassay analyzer.

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One aliquot of each serum sample was spiked with the interfering substance, and another aliquot with the same volume of the solvent (buffer matrix) of the interfering substance (dilution pool). The interfering pool was then diluted into the dilution pool in 10% increments.

The recovery for each sample was calculated by comparison to the reference sample.

Serum Albumin, IgG, IgA, and IgM Interference 4.4.3.

One aliquot of each serum sample was spiked with the interfering substance (lyophilisate). In this case, another aliquot without any additives (since the interfering substance is a lyophilisate) was used as dilution pool. The interfering pool was then diluted into the dilution pool in 14.3% increments.

The deviation or recovery for each sample was calculated by comparison to the reference sample.

Conclusion:

All results met the pre-defined acceptance criteria, demonstrating no interference from endogenous substances up to the levels shown in the table below.

Interferent	Interfering substancemeasured up to	No interferenceseen up to
Intralipid® (Lipemia)	2000 mg/dL	2000 mg/dL
Bilirubin	66 mg/dL	66 mg/dL
Hemoglobin	1000 mg/dL	1000 mg/dL
Rheumatoid Factor	2000 IU/mL	1400 IU/mL
human Serum albumin	7.00 g/dL	7.00 g/dL
human IgG	7.00 g/dL	7.00 g/dL
human IgM	1.00 g/dL	1.00 g/dL
human IgA	1.60 g/dL	1.60 g/dL

Table 6: Summary of Results - Endogenous Interfering Substances

4.5. Biotin

The purpose of this study was to evaluate biotin for potential interference with the parameters measured on the Elecsys Anti-HAV II on the cobas e 601 immunoassay analyzer. Five human serum samples were tested in accordance with CLSI EP07-A2. Negative specimens with biotin

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concentrations up to 100 ng/mL demonstrated ≤11% negative bias in COI values. Biotin concentrations greater than 100 ng/mL lead to higher negative bias and in consequence can lead to false positive Elecsys Anti-HAV II results. Refer to Limitations for additional information on biotin interference.

4.6. Analytical Specificity/Cross-Reactivity

The effect on detection of analyte in the presence of potential cross-reacting antibodies using the Elecsys Anti-HAV II was determined on the cobas e 601 immunoassay analyzer with native human serum and plasma sample pools.

Anti-HAV negative samples containing potential cross-reacting antibodies to other infectious diseases were measured with Elecsys Anti-HAV and Elecsys Anti-HAV II. In total, 10 samples with acute Hepatitis B infection, 10 samples with acute Hepatitis C infection, 10 samples with HIV infection, 10 samples with EBV infection, 10 samples with Anti-CMV antibodies, 10 samples with Anti-HSV antibodies, 10 samples with Toxoplasma Gondii infection, 10 samples with Treponema Pallidum infection, 10 samples with Anti-Mumps/Rubeola antibodies, 10 samples with Anti-Rubella antibodies, 10 samples with Anti-Parvovirus B19 antibodies and 10 samples with ANA Autoimmune antibodies were tested.

No cross-reactivity with other infectious agents was found.

Exogenous Interferences – Drugs 4.7.

The recovery of analyte values in the presence of drugs was determined by comparing values obtained from samples spiked with 18 commonly used pharmaceutical compounds (Acetylcystein, Ampicillin-Na, Ascorbic acid, Ca-Dobesilate, Cyclosporine, Cefoxitin, Doxycycline, Heparin, Levodopa, Methyldopa +1.5, Metronidazole, Phenylbutazone, Tetracycline, Acetylsalicylic Acid, Rifampicin, Acetaminophen, Ibuprofen and Theophylline) with the reference sample (unspiked). Four human serum samples (native human serum pools) were used and tested on the cobas e 601 immunoassay analyzer according to CLSI EP07-A2. The drug concentrations tested correspond at least to the five times maximum daily doses. The four serum samples were divided into aliquots and spiked with the potential interferents. The reference sample without interferent was spiked with the respective amount of solvent only.

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The Elecsys Anti-HAV II concentration (as COI) of the spiked aliquots was determined in threefold determination and compared to the Elecsys Anti-HAV II result determined for the reference aliquot (also in three-fold determination) on one cobas e 601 immunoassay analyzer.

All results met the pre-defined acceptance criteria, demonstrating no interference from the drug substances at the tested concentrations.

Sample Matrix Comparison 4.8.

The recovery of analyte values in the presence of anticoagulants with the Elecsys Anti-HAV II was determined on the cobas e 601 immunoassay analyzer by comparing Elecsys Anti-HAV II results obtained from samples drawn into serum and plasma collection tubes. The samples were spiked with anti-HAV positive samples from individual donors to obtain a range of anti-HAV concentrations. The recovery of each plasma sample to the matching serum sample was calculated. At least 60 serum/plasma pairs were tested for each kind of anticoagulant in single determination (K2-EDTA, K3-EDTA, Na-Heparin, Li-Heparin, Na-Citrate).

The specifications were met for all anti-coagulants, demonstrating that K2-EDTA, K3-EDTA, Na-Heparin, Li-Heparin, and Na-Citrate are acceptable sample types for use with Elecsys Anti-HAV II.

Analytical Method Comparison to Predicate 4.9.

A method comparison of Elecsys Anti-HAV II on the cobas e 601 immunoassay analyzer versus the predicate device, Elecsys Anti-HAV, was conducted according to CLSI EP09-A3.

A total of 215 modified serum samples (≥100 negative and ≥100 positive samples) were measured in singleton on cobas e 601 immunoassay analyzer. The samples were primarily prepared by mixing negative and positive sera from individual donors.

Results for the method comparison between Elecsys Anti-HAV II and Elecsys Anti-HAV met the pre-defined acceptance criteria, which demonstrates equivalency between the candidate and predicate devices.

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Table 7: Analytical Method Comparison Agreement Rates Between Elecsys Anti-HAV II and Elecsys Anti-HAV

Elecsys Anti-HAV II Result	Elecsys Anti-HAV Result
	Reactive	Borderline	Non-Reactive
Reactive	107	2	0
Non-Reactive	0	8	98
Total	107	10	98

The positive percent agreement rate was 100% (107/107), the negative percent agreement rate was 100% (98/98). Of ten samples with results in the borderline zone of the predicate device, Elecsys Anti-HAV, 20% (2/10) were positive and 80% (8/10) were negative by the Elecsys Anti-HAV II.

4.10. Reagent Stability

The reagent stability testing was performed in two different studies:

Study 1: Reagent stability after first opening at 2-8°C (8 weeks) .
Study 2: On-board reagent stability (8 weeks) .

4.10.1. Reagent Stability After First Opening

Elecsys Anti-HAV II reagent kits can be used after first opening for up to 8 weeks when stored at 2-8°C. Reagent stability after first opening for the Elecsys Anti-HAV II assay was tested on one cobas e 601 immunoassay analyzer.

A fresh reagent rackpack was placed on the analyzer and calibrated. Reference values for the samples tested were determined. After initial measurement the kit was removed from the analyzer and kept at 2-8 °C for 35 and 63 days. After 35 and 63 days, the kit was placed on the analyzer again, calibrated, and the test samples were determined. Recovery was calculated based on the result at time-point Day 0. Two control samples and seven samples were tested in duplicate.

All pre-defined acceptance criteria were met, demonstrating reagent stability for 8 weeks after first opening at 2-8°C.

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4.10.2. On-Board Reagent Stability

Elecsys Anti-HAV II reagent kits can be stored on-board the analyzers for up to 8 weeks. A new calibration of the kit kept on-board is recommended every 7 days.

Reagent on-board stability for the Elecsys Anti-HAV II assay was tested on one cobas e 601 immunoassay analyzer. A fresh reagent rackpack was placed on the analyzer and calibrated. All samples were measured on day 0. On day 22, 29, 35, 49 and 63, the same samples were measured with the same reagent kit kept at 20°C ± 3°C (on-board condition) using the calibration curves established on day 0, 22, 29, 35, 49 and 63, respectively. Recovery was calculated based on the result at time-point Day 0. Two control samples were tested in singleton and seven samples were tested in duplicate.

All pre-defined acceptance criteria were met, demonstrating reagent on-board stability for 8 weeks on the cobas e 601 immunoassay analyzer.

4.11. Calibration Stability

To test calibration stability, two studies were completed, including:

Study 1: Lot calibration stability .
Study 2: On-board calibration stability .

4.11.1. Lot Calibration Stability

Calibration of an Elecsys Anti-HAV II reagent lot is recommended every 28 days (4 weeks). During that time period, fresh reagent kits of the same lot can be used without calibration using the calibration curve of the Day 0 reagent kit.

Elecsys Anti-HAV II was calibrated with a fresh reagent kit on day 0 using a cobas e 601 immunoassay analyzer. After 35 days (5 weeks), a new reagent kit of the same lot was used and recovery of samples was determined using the calibration curve of Day 0. Two control samples were tested in singleton and seven human serum samples were tested in duplicate.

All pre-defined acceptance criteria were met, demonstrating renewed calibration is recommended after 4 weeks when using the same reagent lot.

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4.11.2. On-Board Calibration Stability

On-board calibration stability for the Elecsys Anti-HAV II assay was tested on one cobas e 601 immunoassay analyzer. A fresh reagent rackpack was placed on the analyzer and calibrated. All samples were measured on day 0. On day 8, the same samples were measured with the same reagent kit kept at 20°C ± 3°C (on-board condition) using the calibration curves established on day 1. Two control samples were tested in singleton and seven human serum samples were tested in duplicate.

All pre-defined acceptance criteria were met, demonstrating that a calibration can be used for up to one week when using the same lot of reagents stored on the analyzer.

4.12. Sample Stability

To test sample stability, four studies were completed, including:

Study 1: Sample stability at 2-8°C .
Study 2: Sample stability at room temperature (15-25°C) .
Study 3: Sample stability at -15 to -25°C .
Study 4: Freeze/thaw cycles .

4.12.1. Sample Stability at 2-8°C

Six human serum, K2-EDTA, Li-Heparin, and Na-Citrate plasma samples and seven K3-EDTA and Na-Heparin plasma samples were aliquoted and measured fresh (reference value) and after storage at 2-8°C for 14 days. Measurements were performed with three-fold determination on a cobas e 601 immunoassay analyzer and recovery was calculated as percent of the reference value.

All pre-defined acceptance criteria were met, demonstrating the specimens are stable for 14 days at 2-8°C.

4.12.2. Sample Stability at Room Temperature (15-25°C)

Six human serum, K2-EDTA, Li-Heparin, and Na-Citrate plasma samples and seven K3-EDTA and Na-Heparin plasma samples were aliquoted and measured fresh (reference value) and after

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storage at 15-25°C for 6 days. Measurements were performed with three-fold determination on a cobas e 601 immunoassay analyzer and recovery was calculated as percent of the reference value.

All pre-defined acceptance criteria were met, demonstrating the specimens are stable for 6 days at 15-25°C.

4.12.3. Sample Stability at -15 to -25°C

Six human serum, K2-EDTA, Li-Heparin, and Na-Citrate plasma samples and seven K3-EDTA and Na-Heparin plasma samples were aliquoted and measured fresh (reference value) and after storage at -15 to -25℃ for 3 months. Measurements were performed with three-fold determination on a cobas e 601 immunoassay analyzer and recovery was calculated as percent of the reference value.

All pre-defined acceptance criteria were met, demonstrating the specimens are stable for 3 months at -15 to -25°C.

4.12.4. Sample Stability – Freeze/Thaw Cycles

Six human serum, K2-EDTA, Li-Heparin, and Na-Citrate plasma samples and seven K3-EDTA and Na-Heparin plasma samples were aliquoted and measured fresh (reference value) and after five freeze/ thaw cycles. Measurements were performed with three-fold determination on a cobas e 601 immunoassay analyzer and recovery was calculated as percent of the reference value.

All pre-defined acceptance criteria were met, demonstrating the specimens are stable for 5 freeze/thaw cycles.

EXTERNAL (CLINICAL) TESTING 5.

5.1. Reproducibility

Precision results were collected on three different cobas e 601 immunoassay analyzers across three external testing sites utilizing a single lot of Elecsys Anti-HAV II reagent. Four human serum pools and two Elecsys Anti-HAV II PreciControl materials were tested in three replicates

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per run, two runs per day for a 5-day format according to CLSI EP05-A3. All testing was performed on the same analyzer with a new rackpack for each day. Repeatability, withinlaboratory, and reproducibility were calculated as SD and CV values.

Ten dummy specimens were tested in each run, as well as between runs, so as to separate the runs within each day. The data were evaluated by Component of Variance Analysis.

					Repeatability		BetweenRun		BetweenDay		BetweenSite		Reproducibility
SpecimenMaterial	N	Mean	Median	Min	Max	SD(95%CI)	%CV(95%CI)	SD	%CV	SD	%CV	SD	%CV	SD(95%CI)	%CV(95%CI)
HSP 01	90	0.813	0.814	0.764	0.856	0.010(0.012)	1.3(1.5)	0.016	1.9	0.000	0.0	0.011	1.3	0.022(0.031)	2.7(3.8)
HSP 02	90	0.904	0.908	0.846	0.953	0.012(0.014)	1.3(1.5)	0.016	1.8	0.000	0.0	0.014	1.5	0.024(0.036)	2.7(4.0)
HSP 03	90	1.009	1.010	0.934	1.070	0.018(0.021)	1.7(2.1)	0.013	1.3	0.004	0.4	0.016	1.6	0.027(0.041)	2.7(4.1)
HSP 04	90	0.449	0.450	0.408	0.478	0.005(0.006)	1.1(1.3)	0.011	2.5	0.000	0.0	0.007	1.5	0.014(0.020)	3.1(4.4)
PC A-HAV2_L1	90	1.299	1.300	1.230	1.350	0.018(0.022)	1.4(1.7)	0.009	0.7	0.010	0.7	0.018	1.4	0.029(0.046)	2.2(3.5)
PC A-HAV2_L2	90	0.376	0.375	0.347	0.410	0.006(0.007)	1.6(1.9)	0.013	3.3	0.000	0.0	0.003	0.9	0.014(0.018)	3.8(4.8)

Table 8: Results - Elecsys Anti-HAV II and PreciControls Anti-HAV II on cobas e 601 Immunoassay Analyzer (all sites combined)

One-sided upper 95% confidence interval displayed

5.2. Method Comparison Versus Predicate

The Elecsys Anti-HAV II was performed at each of the three clinical laboratories with a single lot of reagent to assess the performance of the assay in a testing environment that most closely resembles that of the final user. The objective of this study was to provide data for US- and non-US-obtained specimens by testing the Elecsys Anti-HAV II on the cobas e 601 immunoassay analyzer and comparing the results to an FDA-cleared predicate assay. The Elecsys Anti-HAV assay was used as the predicate device for comparison at each of the three laboratories. The study included specimens from patients in the following cohorts: routine HAV testing, hospitalized, increased risk, symptomatic, characterized acute HAV infected, and pediatric.

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	Elecsys Anti-HAV Assay Results (all sites combined)
Elecsys Anti-HAVII Results	Reactive$\geq$ 22 IU/L	Border 18.0 $\leq$ IU/L < 22.0	Non-Reactive$<$ 18.0 IU/L	Total
Reactive COI $\leq$ 1.0	501	6	16	523
Non-ReactiveCOI >1.0	0	1	437	438
Total	501	7	453	961

Table 9: Method Comparison Results (all sites combined)

	Absolute	Relative %	95 % CI
PPAc)	501/502	99.80	98.90; 99.99
NPAd)	437/459	95.21	92.83; 96.97

c) PPA = Positive Percent Agreement

NPA = Negative Percent Agreement d)

Positive percent agreement and negative percent agreement with their respective 95th percentile confidence interval (CI) for each cohort are summarized in the following table. The lower end of the 95% confidence intervals for PPA ranged from 90.45% in the hospitalized cohort to 97.18% in the symptomatic cohort. The lower end of the ≥ 95% confidence intervals for NPA ranged from 52.36% in the pediatric cohort to 93.04% in the hospitalized cohort. For the pediatric cohort, there were 14 specimens negative by both the predicate Elecsys Anti-HAV assay and the Elecsys Anti-HAV II assay. There were 4 discordant results, of which 2 were borderline with the predicate assay but positive with the Elecsys Anti-HAV II assay and 2 were negative with the predicate assay but positive with the Elecsys Anti-HAV II assay. Discordant results were counted against Elecsys Anti-HAV II when calculating agreement (77.8%). The lower bound of the 95% CI for the NPA in the pediatric population is influenced by the small number of specimens (n = 18). The overall percentages for all cohorts combined met the expected performance of a lower bound of the 95% confidence interval of ≥ 90% for the agreement rates (positive and negative) versus the predicate assay (Elecsys Anti-HAV assay). The lower limit of the confidence interval for the PPA was 98.90%, and the lower limit of the confidence interval for NPA was 92.83%.

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Summary of the percent agreements for various specimen cohorts: Elecsys Anti-HAV IIe) assayversus the predicate (Elecsys Anti-HAV assay)f)
Cohort	PPA		NPA
	PPA (x/n)	95 % CI	NPA (x/n)	95 % CI
Routine HAV testing	100 (91/91)	(96.03, 100)	94.50 (103/109)	(88.40, 97.95)
Hospitalized	98.21 (55/56)	(90.45, 99.95)	97.22 (140/144)	(93.04, 99.24)
Increased risk for hepatitis	100 (119/119)	(96.95, 100)	94.25 (82/87)	(87.10, 98.11)
Symptomatic	100 (129/129)	(97.18, 100)	96.70 (88/91)	(90.67, 99.31)
Characterized acute HAV	100 (65/65)	(94.48, 100)	100 (10/10)	(69.15, 100)
Pediatric	100 (42/42)	(91.59, 100)	77.78 (14/18)	(52.36, 93.59)
Overall	99.80 (501/502)	(98.90, 99.99)	95.21 (437/459)	(92.83, 96.97)

Table 10: Percent Agreements for the Various Specimen Cohorts

e) Cutoff of 1.0 COI used for Elecsys Anti-HAV II assay

f) Specimens with results in the borderline range of (18.0 ≤ IU/L < 22.0) for Elecsys Anti-HAV assay were counted as discordant with the Elecsys Anti-HAV II assay

Pre- and Post-HAV Vaccination 5.3.

Specimens from 49 subjects that were collected both pre- and post-HAV vaccination were evaluated by the Elecsys Anti-HAV II assay and the predicate assay (Elecsys Anti-HAV). The post-vaccination specimens were obtained at least 4 weeks, but not more than 10 weeks, after the completion of the vaccine regimen. Three HAV vaccines, which are currently licensed in the U.S., were used. No discrepant results were observed.

5.4. Prevalence Study

The Elecsys Anti-HAV II assay was used to evaluate the prevalence of HAV antibodies in an apparently healthy population (presumed normal, healthy individuals without symptoms derived by a self-reporting health questionnaire). A statistically significant number of subjects were collected in a presumed "high prevalence" region, the Western United States, and a presumed

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"low prevalence" region, the Eastern United States. Testing based on the predicate assay was not required.

The collection site representing the Eastern US recruited 431 subjects. Of the 431 subject, 31 were classified as screen failures and therefore did not complete the informed consent process. A total of 400 subjects satisfied the inclusion/exclusion criteria and proved acceptable specimens for analysis. The collection site representing the Western US recruited 427 subjects. Of the 427 subjects, 24 were classified as screen failures and therefore did not complete the informed consent process. An additional three subjects were dropped from enrollment. A subject was allowed to be dropped from enrollment if defined conditions were met after the screening process was completed and the informed consent process was successfully completed. Both collection sites were open to the recruitment/enrollment of subjects for Prevalence cohort only. Each site was instructed to collect 400 evaluable specimens, which was accomplished.

The results were consistent with a higher prevalence of reactive HAV results reported in the Western US, 53.75%, versus the Eastern US 29.50%.

6. ADDITIONAL INFORMATION

The Elecsys Anti-HAV II is intended to be used with the following calibrators and controls:

PreciControl Anti-HAV II .
Anti-HAV II Call and Anti-HAV II Cal2, which is included in the kit .

CONCLUSIONS 7.

The information provided in this 510(k) Premarket Notification supports a determination of substantial equivalence for the Elecsys Anti-HAV II. The data from the non-clinical and clinical tests demonstrate that the device is as safe, as effective, and performs as well as the predicate device.

Regulation Number and Section

§ 866.3310 Hepatitis A virus (HAV) serological assays.

(a)
Identification. HAV serological assays are devices that consist of antigens and antisera for the detection of hepatitis A virus-specific IgM, IgG, or total antibodies (IgM and IgG), in human serum or plasma. These devices are used for testing specimens from individuals who have signs and symptoms consistent with acute hepatitis to determine if an individual has been previously infected with HAV, or as an aid to identify HAV-susceptible individuals. The detection of these antibodies aids in the clinical laboratory diagnosis of an acute or past infection by HAV in conjunction with other clinical laboratory findings. These devices are not intended for screening blood or solid or soft tissue donors.(b)
Classification. Class II (special controls). The special control is “Guidance for Industry and FDA Staff: Class II Special Controls Guidance Document: Hepatitis A Virus Serological Assays.” See § 866.1(e) for the availability of this guidance document.