ANTI-HAV
Device Facts
| Record ID | K100903 |
|---|---|
| Device Name | ANTI-HAV |
| Applicant | Roche Professional Diagnostics |
| Product Code | LOL · Microbiology |
| Decision Date | Oct 5, 2010 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.3310 |
| Device Class | Class 2 |
| Attributes | Pediatric |
Intended Use
The Roche Elecsys Anti-HAV immunoassay is used for the in vitro qualitative detection of total antibodies (IgM and IgG) to hepatitis A virus in human serum and plasma (K₂-EDTA). The assay is intended for use as an aid in the laboratory diagnosis of past or acute/recent hepatitis A infection. Assay results, in conjunction with other laboratory results and clinical information, may be used to provide presumptive evidence of infection with hepatitis A virus in persons with signs or symptoms of hepatitis and in persons at risk for hepatitis A infection, or used as an aid to determine the presence of antibody response to HAV in vaccine recipients. The electrochemiluminescence immunoassay “ECLIA” is intended for use on Elecsys and cobas e immunoassay analyzers. Elecsys PreciControl Anti-HAV is used for the quality control of the Elecsys Anti-HAV immunoassay on the Elecsys and cobas e immunoassay analyzers. This assay is not intended for screening blood or solid or soft tissue donors. Assay performance characteristics have not been established for immunocompromised or immunosuppressed patients. The users are responsible for establishing their own assay performance characteristics in these populations. Caution: U.S. Federal Law restricts this device to sale by or on the order of a physician.
Device Story
The Elecsys Anti-HAV assay is an in vitro diagnostic test for total anti-HAV antibodies in human serum or K2-EDTA plasma. It utilizes a competitive electrochemiluminescence immunoassay (ECLIA) format. Sample antibodies compete with biotinylated and ruthenylated anti-HAV monoclonal antibodies for a limited amount of HAV antigen. Immune complexes are captured on streptavidin-coated magnetic microparticles. Upon voltage application to the electrode, the ruthenium complex emits chemiluminescence measured by a photomultiplier. The signal is inversely proportional to the anti-HAV concentration. The device is used on Roche Elecsys and cobas e immunoassay analyzers in clinical laboratories. Results are interpreted qualitatively against a 20 IU/L cutoff. The assay aids clinicians in diagnosing past or recent hepatitis A infection and assessing vaccine response. It provides presumptive evidence of infection when used with other clinical information.
Clinical Evidence
Clinical performance was evaluated in a multi-center U.S. study of 1050 samples (prospective and retrospective) including symptomatic patients, routine testing, hospitalized patients, acute HAV cases, and pediatric/adolescent subjects. Overall positive percent agreement (PPA) was 98.12% (95% CI: 96.46-99.14%) and negative percent agreement (NPA) was 97.37% (95% CI: 95.70-98.52%) compared to the predicate. Seroconversion sensitivity was demonstrated using 3 panels. Prevalence studies were conducted in high and low-risk U.S. regions.
Technological Characteristics
Competitive ECLIA format; uses cell culture-derived HAV antigen, biotinylated/ruthenylated monoclonal antibodies, and streptavidin-coated magnetic microparticles. Signal generation via electrochemiluminescence (Tris(2,2'-bipyridyl)ruthenium(II)-complex). Analyzers: Elecsys 2010, cobas e 411, MODULAR ANALYTICS E170, cobas e 601. Standardized against NIBSC 97/646. Qualitative interpretation based on 20 IU/L cutoff. Reagent barcode contains master curve data.
Indications for Use
Indicated for the qualitative detection of total (IgM and IgG) anti-HAV antibodies in human serum and K2-EDTA plasma. Used as an aid in the diagnosis of past or acute/recent hepatitis A infection in symptomatic individuals, persons at risk for hepatitis A, or to determine antibody response in vaccine recipients. Not for blood/tissue donor screening. Performance not established for immunocompromised/immunosuppressed patients.
Regulatory Classification
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.
Special Controls
*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.
Predicate Devices
- Abbott AxSym HAVAB 2.0 (P780012/S009)
Related Devices
- K190428 — Elecsys Anti-HAV II · Roche Diagnostics · Aug 13, 2019
- K093955 — ANTI-HAV IGM · Roche Diagnostics · Jun 22, 2010
Submission Summary (Full Text)
{0}------------------------------------------------
## 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
- A. 510(k) Number: K100903
- B. Purpose for Submission: Clearance of New Device
- C. Measurand: Total Antibody to Hepatitis A Virus in serum and heparinized plasma
- D. Type of Test: Electrochemiluminescence immunoassay (ECLIA)
- E. Applicant: Roche Diagnostics
- F. Proprietary and Established Names: Elecsys® Anti-HAV Assay Elecsys® PreciControl anti-HAV Assay
#### G. Regulatory Information:
| 1. Regulation section: | 21 CFR §866.3310, Hepatitis A virus Serological Assays<br>21 CFR §862.1660, Quality Control Material |
|------------------------|-------------------------------------------------------------------------------------------------------------|
| 2. Classification: | Class II |
| 3. Product code: | LOL (Hepatitis A Test – antibody and IgM Antibody)<br>JJX (Quality control material, assayed and unassayed) |
| 4. Panel: | Microbiology (83) |
### H. Intended Use:
- 1. Intended use(s):
The Roche Elecsys Anti-HAV immunoassay is used for the in vitro qualitative detection of total antibodies (IgM and IgG) to hepatitis A virus in human serum and plasma (K2-EDTA). The assay is intended for use as an aid in the laboratory diagnosis of past or acute/recent hepatitis A infection.
Assay results, in conjunction with other laboratory results and clinical information, may be used to provide presumptive evidence of infection with hepatitis A virus in persons with signs or symptoms of hepatitis and in persons at risk for hepatitis A infection, or used as an aid to determine the presence of antibody response to HAV in vaccine recipients.
The electrochemiluminescence immunoassay "ECLIA" is intended for use on Elecsys and cobas e immunoassav analyzers.
Elecsys PreciControl Anti-HAV is used for the quality control of the Elecsys Anti-HAV immunoassay on the Elecsys and cobas e immunoassay analyzers.
{1}------------------------------------------------
This assay is not intended for screening blood or solid or soft tissue donors. Assay performance characteristics have not been established for immunocompromised or immunosuppressed patients. The users are responsible for establishing their own assay performance characteristics in these populations.
Caution: U.S. Federal Law restricts this device to sale by or on the order of a physician.
- 2. Indication(s) for use:
Same as Intended Use
- 3. Special conditions for use statement(s):
For prescription use only
- 4. Special instrument requirements:
Elecsys 2010 and cobas e 411 analyzers; MODULAR ANALYTICS E170 and cobas e 601 analyzers
## I. Device Description:
The Elecsys anti-HAV test is a qualitative assay based on electrochemiluminescence immunoassay "ECLIA" technology. The Elecsys anti-HAV test utilizes a competitive immunoassay format in which sample anti-HAV antibody competes with biotinylated and ruthenvlated anti-HAV monoclonal antibodies for a limited amount of cell culture-derived HAV antigen. The sample antibody and the HAV antigen react in the first incubation. The biotinylated antibodies and ruthenium complex® -labeled antibodies specific for HAV antigen are added in the second incubation together with streptavidin-coated magnetic microparticles. The unbound HAV antigen reacts with the modified antibodies and the resulting immune complexes are bound to the solid phase through a biotinstreptavidin interaction. If all HAV antigens are complexed by sample anti-HAV antibody during the first incubation, no modified/labeled immune complexes are formed and captured during the second incubation.
Following the second incubation, the reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are removed by elution with ProCell. Application of a voltage to the electrode induces chemiluminescent emission from the captured immune complexes which is measured by a photomultiplier. The level of signal detected by the system decreases as the concentration of the anti-HAV antibody target present in a patient sample increases.
{2}------------------------------------------------
Results are determined via a calibration curve which is generated by 2-point calibration on the instrument and a master curve provided via the reagent barcode. The calibration process converts the output so that low levels of sample anti-HAV antibodies are expressed by low output and high levels of antibody are expressed by high output. These outputs are finally interpreted on a qualitative basis around the established cut-off output.
a) Tris(2,2'-bipyridyl)ruthenium(II)-complex (Ru(bpy)) 3 2+
# J. Substantial Equivalence Information:
- 1. Predicate device name(s): Abbott AxSym HAVAB® 2.0
- Predicate Numbers (s): 2.
- 3. Comparison with predicate:
| Similarities | | |
|----------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Anti-HAV Immunoassay Comparison | | |
| Feature | Elecsys Anti-HAV Assay | Predicate Device<br>Abbott Axsym HAVAB 2.0<br>Assay (P780012/S009) |
| Intended Use | The Roche Elecsys Anti-HAV<br>immunoassay is used for the in vitro<br>qualitative detection of total antibodies<br>(IgM and IgG) to hepatitis A virus in<br>human serum and plasma (K2-EDTA).<br>The assay is intended for use as an aid in<br>the laboratory diagnosis of past or<br>acute/recent hepatitis A infection.<br>Assay results, in conjunction with other<br>laboratory results and clinical information,<br>may be used to provide presumptive<br>evidence of infection with hepatitis A<br>virus in persons with signs or symptoms<br>of hepatitis and in persons at risk for<br>hepatitis A infection, or used as an aid to<br>determine the presence of antibody<br>response to HAV in vaccine recipients.<br>The electrochemiluminescence<br>immunoassay “ECLIA” is intended for<br>use on Elecsys and cobas e immunoassay<br>analyzers. | Immunoassay for the qualitative<br>detection of total antibody to<br>hepatitis A virus (anti-HAV) in<br>human serum or plasma<br>(potassium EDTA, sodium<br>heparin, sodium citrate, or<br>lithium heparin). A test for anti-<br>HAV is indicated as an aid in the<br>laboratory diagnosis of previous<br>or ongoing hepatitis A viral<br>infection or in the identification<br>of HAV-susceptible individuals<br>for vaccination.<br>Assay results, in conjunction<br>with other laboratory results and<br>clinical information, may be<br>used to provide presumptive<br>evidence of infection with<br>hepatitis A virus in persons with<br>signs or symptoms of hepatitis<br>and in persons at risk for<br>hepatitis A infection. |
| Indications for<br>Use | Same | Same |
| Sample Type | Human serum and plasma | Same |
| Anti-HAV Immunoassay Comparison | | |
| Features | Elecsys Anti-HAV Assay | Predicate Device<br>Abbott Axsym HAVAB 2.0 Assay<br>(P780012/S009) |
| Detection<br>Protocol | Electrochemiluminescence<br>immunoassay (ECLIA) | Microparticle Enzyme Immunoassay<br>(MEIA) |
| Traceability/<br>Standardization | Standardized against the "Second<br>International Standard for Anti-<br>Hepatitis A, Immunoglobulin,<br>Human, NIBSC code: 97/646" of<br>the NIBSC (National Institute for<br>Biological Standards and Control). | Not Given |
| Interpretation of<br>Results | ≥ 22.0 IU/L Reactive<br>18.0 ≤ IU/L< 22.0 Equivocal<br><18.0 IU/L Negative | 0.000 to 1.000 Reactive<br>1.001 to 1.200 Grayzone<br>1.201 to 3.000 Nonreactive<br>>3.000 Not applicable |
| Calibration<br>Interval | Once per reagent lot and<br>• After 1 month (28 days) when<br>using the same reagent lot<br>• After 7 days (when using the<br>same reagent kit on the analyzer)<br>• As required: e.g., quality control<br>findings outside the specified<br>limits | A minimum of two replicates of the<br>AxSYM HAFVAB 2.0 Index Calibrator<br>must be tested. A single sample of both<br>the Negative and Positive Controls must<br>be tested as a means of evaluating the<br>assay calibration. Once the calibration is<br>accepted and stored, all subsequent<br>samples may be tested without further<br>calibration unless one or more of the<br>following occur:<br>• A reagent pack with a new lot number<br>is used<br>• Either of the AxSYM HAVAB 2.0<br>Control values is out of its specified<br>range<br>• The MEIA Optics Verification Update<br>has been performed |
| Controls | Elecsys PreciControl Anti-HAV | Abbott AxSYM HAVAB 2.0 Controls |
{3}------------------------------------------------
# Differences
# K. Standard/Guidance Document referenced (if applicable):
CLSI EP5-A2, "Evaluation of Precision Performance of Quantitative Measurement Methods"
{4}------------------------------------------------
CLSI EP17-A, "Protocols for Determination of Limits of Detection" Class II Special Controls Guidance Document: Hepatitis A Virus Serological Assays http://www.fda.gov/downloads/MedicalDevices/DeviceRegulationandGuidance/Guidan ceDocuments/ucm071055.pdf
# L. Test Principle:
The Elecsys Anti-HAV immunoassay utilizes a competitive immunoassay format employing cell culture derived HAV antigen and a monoclonal antibody (modified by biotinylation or ruthenylation) in competition to the polyclonal sample antibody. Capture of formed immune complexes from the reaction mixture is based on biotin in the immune complex binding to streptavidin-coated magnetic microparticles which are collected on a measuring cell electrode. Signal generation is triggered by the application of a voltage to the electrode (electrochemiluminescence technology). The level of signal count detected by the system decreases as the concentration of the anti-HAV total antibody target present in a patient sample increases. Results are determined via a calibration curve which is instrument-specifically generated by 2-point calibration and a master curve provided via the reagent barcode
# M. Performance Characteristics (if/when applicable):
# 1. Analytical performance:
# a. Precision/Reproducibility:
Precision and Reproducibility of the Elecsys Anti-HAV assay was evaluated on Elecsys 2010 analyzer and MODULAR ANALYTICS E170 analyzer using Elecsys reagents, three human sera and controls, according to the CLSI EP15-A2/EP5-A2.
For precision study, three human sera (high negative, low positive and moderately positive) and controls (PreciControl Anti-HAV 1 and PreciControl Anti-HAV 2) were tested in replicates of 2 in 2 runs per day for 20 days with one reagent lot spanning at least two cycles of calibration using one Elecsys 2010 analyzer and one Modular Analytics E170 analyzer. Results are presented below.
| Sample | Mean<br>(IU/L) | Repeatability | | Intermediate Precision<br>(within-laboratory<br>precision) | | n |
|---------------|----------------|---------------|---------------------|------------------------------------------------------------|---------------------|----|
| | | SD<br>(IU/L) | CV (%)<br>(UCL 95%) | SD<br>(IU/L) | CV (%)<br>(UCL 95%) | |
| PC Anti-HAV 1 | 12.3 | 0.221 | 1.8 (2.2) | 0.378 | 3.1 (3.7) | 80 |
| PC Anti-HAV 2 | 29.2 | 0.947 | 3.2 (4.0) | 1.325 | 4.5 (5.3) | 80 |
| HSP 1 | 17.6 | 0.534 | 3.0 (3.7) | 0.586 | 3.3 (3.8) | 80 |
| HSP 2 | 21.9 | 0.538 | 2.5 (3.0) | 0.667 | 3.0 (3.6) | 80 |
| HSP3 | 49.3 | 1.655 | 3.4 (4.1) | 2.261 | 4.6 (5.4) | 80 |
- Precision on Elecsys® 2010 ●
{5}------------------------------------------------
| Sample | Mean<br>(IU/L) | Repeatability | | Intermediate Precision<br>(within-laboratory<br>precision) | | |
|---------------|----------------|---------------|---------------------|------------------------------------------------------------|---------------------|----|
| | | SD<br>(IU/L) | CV (%)<br>(UCL 95%) | SD<br>(IU/L) | CV (%)<br>(UCL 95%) | n |
| PC Anti-HAV 1 | 12.54 | 0.188 | 1.5 (1.8) | 0.688 | 5.5 (7.1) | 80 |
| PC Anti-HAV 2 | 32.12 | 0.724 | 2.3 (2.8) | 1.849 | 5.8 (7.4) | 80 |
| HSP 1 | 18.2 | 0.459 | 2.5 (3.1) | 0.816 | 4.5 (5.6) | 80 |
| HSP 2 | 23 | 0.254 | 1.1 (1.4) | 1.016 | 4.4 (5.9) | 80 |
| HSP3 | 55.37 | 1.088 | 2.0 (2.4) | 2.374 | 4.3 (5.3) | 80 |
#### Precision on MODULAR® ANALYTICS E170 .
PC = PreControl
UCL = Upper Confidence Limit
For reproducibility study, three human sera (high negative, low positive and moderately positive) and controls (PreciControl Anti-HAV 1 and PreciControl Anti-HAV 2) were tested in replicates of 3 in 2 runs per day for 5 days with three Elecsys 2010 analyzers and three Modular Analytics E170 analyzers. Results are presented below.
| Clinical | Sample | N | Mean<br>IU/L | Repeatability | | Between Run | | Between Day | | Within Site<br>Reproducibility | |
|----------|---------------|----|--------------|---------------|------|-------------|------|-------------|------|--------------------------------|------|
| Site | | | | SD | % CV | SD | % CV | SD | % CV | SD | % CV |
| Site 1 | HSP 01 | 30 | 22.629 | 0.523 | 2.3 | 0.275 | 1.2 | 0.176 | 0.8 | 0.616 | 2.7 |
| | HSP 02 | 30 | 49.903 | 2.469 | 4.9 | 0.000 | 0.0 | 0.123 | 0.2 | 2.472 | 5.0 |
| | HSP 03 | 30 | 18.287 | 0.527 | 2.9 | 0.052 | 0.3 | 0.264 | 1.4 | 0.592 | 3.2 |
| | PC A-<br>HAV1 | 30 | 12.678 | 0.321 | 2.5 | 0.000 | 0.0 | 0.201 | 1.6 | 0.379 | 3.0 |
| | PC A-<br>HAV2 | 30 | 29.198 | 0.707 | 2.4 | 0.516 | 1.8 | 0.505 | 1.7 | 1.011 | 3.5 |
| Site 2 | HSP 01 | 30 | 23.196 | 0.302 | 1.3 | 0.452 | 2.0 | 0.000 | 0.0 | 0.544 | 2.3 |
| | HSP 02 | 30 | 50.965 | 0.688 | 1.4 | 0.55 | 1.1 | 0.000 | 0.0 | 0.881 | 1.7 |
| | HSP 03 | 30 | 18.499 | 0.417 | 2.3 | 0.336 | 1.8 | 0.000 | 0.0 | 0.535 | 2.9 |
| | PC A-<br>HAV1 | 30 | 12.942 | 0.278 | 2.2 | 0.309 | 2.4 | 0.000 | 0.0 | 0.416 | 3.2 |
| | PC A-<br>HAV2 | 30 | 30.814 | 0.376 | 1.2 | 0.441 | 1.4 | 0.000 | 0.0 | 0.58 | 1.9 |
| Site 3 | HSP 01 | 30 | 22.634 | 0.371 | 1.6 | 0.000 | 0.0 | 0.266 | 1.2 | 0.456 | 2.0 |
| | HSP 02 | 30 | 48.03 | 1.356 | 2.8 | 1.451 | 3 | 0.000 | 0.0 | 1.985 | 4.1 |
| | HSP 03 | 30 | 17.892 | 0.398 | 2.2 | 0.268 | 1.5 | 0.292 | 1.6 | 0.561 | 3.1 |
| | PC A-<br>HAV1 | 30 | 12.515 | 0.248 | 2.0 | 0.106 | 0.9 | 0.372 | 3.0 | 0.459 | 3.7 |
| | PC A-<br>HAV2 | 30 | 29.46 | 1.144 | 3.9 | 0.962 | 3.3 | 0.503 | 1.7 | 1.578 | 5.4 |
Elecsys anti-HAV: Within Site Reproducibility: Elecsys® 2010 Analyzer
{6}------------------------------------------------
| Clinical<br>Site | Sample | N | Mean<br>IU/L | Repeatability | | Between Run | | Between Day | | Within Site<br>Reproducibility | |
|------------------|---------------|----|--------------|---------------|------|-------------|------|-------------|------|--------------------------------|------|
| | | | | SD | % CV | SD | % CV | SD | % CV | SD | % CV |
| Site 1 | HSP 01 | 30 | 23.433 | 0.237 | 1.0 | 0.419 | 1.8 | 0.000 | 0.0 | 0.481 | 2.1 |
| | HSP 02 | 30 | 51.725 | 0.454 | 0.9 | 0.707 | 1.4 | 0.000 | 0.0 | 0.84 | 1.6 |
| | HSP 03 | 30 | 18.918 | 0.158 | 0.8 | 0.167 | 0.9 | 0.135 | 0.7 | 0.266 | 1.4 |
| | PC A-<br>HAV1 | 30 | 13.205 | 0.197 | 1.5 | 0.211 | 1.6 | 0.132 | 1.0 | 0.317 | 2.4 |
| | PC A-<br>HAV2 | 30 | 31.477 | 0.339 | 1.1 | 0.787 | 2.5 | 0.000 | 0.0 | 0.857 | 2.7 |
| Site 2 | HSP 01 | 30 | 22.541 | 0.705 | 3.1 | 0.090 | 0.4 | 0.231 | 1.0 | 0.747 | 3.3 |
| | HSP 02 | 30 | 51.871 | 1.344 | 2.6 | 0.000 | 0.0 | 0.302 | 0.6 | 1.378 | 2.7 |
| | HSP 03 | 30 | 18.455 | 0.642 | 3.5 | 0.000 | 0.0 | 0.355 | 1.9 | 0.733 | 4 |
| | PC A-<br>HAV1 | 30 | 12.676 | 0.695 | 5.5 | 0.000 | 0.0 | 0.627 | 4.9 | 0.936 | 7.4 |
| | PC A-<br>HAV2 | 30 | 31.438 | 0.733 | 2.3 | 0.231 | 0.7 | 0.000 | 0.0 | 0.769 | 2.4 |
| Site 3 | HSP 01 | 30 | 23.946 | 0.288 | 1.2 | 0.22 | 0.9 | 0.000 | 0.0 | 0.363 | 1.5 |
| | HSP 02 | 30 | 52.166 | 0.299 | 0.6 | 0.829 | 1.6 | 0.000 | 0.0 | 0.881 | 1.7 |
| | HSP 03 | 30 | 19.426 | 0.203 | 1.0 | 0.336 | 1.7 | 0.000 | 0.0 | 0.392 | 2.0 |
| | PC A-<br>HAV1 | 30 | 12.974 | 0.205 | 1.6 | 0.225 | 1.7 | 0.000 | 0.0 | 0.304 | 2.3 |
| | PC A-<br>HAV2 | 30 | 31.236 | 0.241 | 0.8 | 1.038 | 3.3 | 0.000 | 0.0 | 1.066 | 3.4 |
Elecsys anti-HAV: Within Site Reproducibility: MODULAR® ANALYTICS E170
Elecsys anti-HAV: Between Site Reproducibility: Elecsys® 2010 Analyzer
| | N | Mean<br>IU/L | Repeatability | | Between Run | | Between Day | | Between Site | | Reproducibility | |
|---------------|----|--------------|---------------|------|-------------|------|-------------|------|--------------|------|-----------------|------|
| Sample | | | SD | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV |
| HSP 01 | 90 | 22.819 | 0.409 | 1.8 | 0.3 | 1.3 | 0.000 | 0.0 | 0.304 | 1.3 | 0.591 | 2.6 |
| HSP 02 | 90 | 49.633 | 1.674 | 3.4 | 0.697 | 1.4 | 0.000 | 0.0 | 1.462 | 2.9 | 2.33 | 4.7 |
| HSP 03 | 90 | 18.226 | 0.451 | 2.5 | 0.25 | 1.4 | 0.166 | 0.9 | 0.276 | 1.5 | 0.608 | 3.3 |
| PC A-<br>HAV1 | 90 | 12.712 | 0.284 | 2.2 | 0.179 | 1.4 | 0.22 | 1.7 | 0.176 | 1.4 | 0.438 | 3.4 |
| PC A-<br>HAV2 | 90 | 29.824 | 0.807 | 2.7 | 0.68 | 2.3 | 0.394 | 1.3 | 0.808 | 2.7 | 1.386 | 4.6 |
{7}------------------------------------------------
| Sample | N | Mean<br>IU/L | Repeatability | | Between Run | | Between Day | | Between Site | | Reproducibility | |
|---------------|----|--------------|---------------|------|-------------|------|-------------|------|--------------|------|-----------------|------|
| | | | SD | % CV | SD | % CV | SD | % CV | SD | % CV | SD | % CV |
| HSP 01 | 90 | 23.307 | 0.46 | 2 | 0.278 | 1.2 | 0.000c | 0.0 | 0.7 | 3 | 0.883 | 3.8 |
| HSP 02 | 90 | 51.921 | 0.837 | 1.6 | 0.542 | 1 | 0.000c | 0.0 | 0.168 | 0.3 | 1.011 | 1.9 |
| HSP 03 | 90 | 18.933 | 0.399 | 2.1 | 0.148 | 0.8 | 0.17 | 0.9 | 0.472 | 2.5 | 0.658 | 3.5 |
| PC A-<br>HAV1 | 90 | 12.952 | 0.433 | 3.3 | 0.000c | 0 | 0.368 | 2.8 | 0.193 | 1.5 | 0.6 | 4.6 |
| PC A-<br>HAV2 | 90 | 31.384 | 0.487 | 1.6 | 0.764 | 2.4 | 0.000c | 0 | 0.000c | 0 | 0.906 | 2.9 |
Elecsys anti-HAV: Between Site Reproducibility: MODULAR® ANALYTICS E170
# b. Linearity/assay reportable range:
The qualitative assay has a quantitative measurement. The detectable range is from 6 IU/L to 60 IU/L.
- Traceability, Stability, Expected values (controls, calibrators, or methods): C.
Traceability: This method has been standardized against the "Second International Standard for Anti-Hepatitis A, Immunoglobulin, Human, NIBSC code: 97/646" of the NIBSC (National Institute for Biological Standards and Control).
Controls:
PC A-HAV 1: anti-HAV antibodies (human), with approximately 12 IU/L
PC A-HAV 2: anti-HAV antibodies (human), with approximately 30 IU/L
### Calibrators:
Calibration of the Elecsys anti-HAV is performed by using the 2 calibrators provided with the reagent kit. Calibrator 1 is an anti-HAV negative serum; Calibrator 2 is an anti-HAV positive serum with approx. 46 IU/L.
A reagent lot specific master standard curve is provided on the barcode of each reagent kit. The calibrator master standard curve is established by diluting an anti-HAV positive human serum with an analyte free human serum matrix. The concentration value was assigned to it by reading the concentration from the reference calibrator curve using four E2010 and four E170 instruments with 2 runs and 2 replicates per run and per instrument. The master curve is stored in the barcode and is automatically read in by the instrument when the barcode is scanned for the new reagent.
The calibration must be performed once per reagent lot. Acceptance criteria for the assigned values are < 20 IU/L for Cal 1 and > 40 IU/L for Cal 2. Recalibration is recommended if the same reagent lot was used more than 1 month (28 days), or if the same reagent kit has been left on the analyzer for 7 days.
{8}------------------------------------------------
# Stability:
| Reagents: |
|-----------|
|-----------|
| unopened at 2-8 °C | up to the stated expiration date |
|-------------------------------------------------------------------------------|-------------------------------------------------------------------------------------|
| M, R1, R2 after opening at 2-8 °C | 8 weeks |
| on Elecsys® 2010 and cobas e® 411<br>at<br>20-25 °C | 8 weeks |
| | on MODULAR® ANALYTICS E170 and cobas e® 601 at 20-25 °C |
| Reconstituted calibrators at -20 °C | 12 weeks |
| Reconstituted calibrators at 2-8 °C | 2 weeks |
| Reconstituted calibrators on<br>Elecsys® 2010 and cobas e® 411 at<br>20-25 °C | up to 6 hours |
| | Reconstituted calibrators on<br>MODULAR® ANALYTICS E170<br>and cobas e® 601 at 32°C |
# PreciControls:
| unopened at 2-8 °C | up to the stated expiration date |
|------------------------------------------------------------|----------------------------------|
| after opening at 2-8 °C | 8 weeks |
| After opening at -20 °C | 3 months (freeze only once) |
| on Elecsys® 2010 and cobas e® 411<br>at 20-25 °C | up to 7 hours |
| on MODULAR® ANALYTICS<br>E170 and cobas e® 601 at 20-25 °C | up to 7 hours |
# Samples:
Sample stability was determined using different sample materials e.g. Serum, Serum with separating gel, K2-EDTA Plasma. Four negative samples, one sample around cutoff, and three low positive samples were tested and compared with fresh reference materials.
| Samples stored at -20°C | up to 3 months |
|--------------------------------------------------|----------------|
| Samples stored at 2-8°C | Up to 7 days |
| Freeze and thaw cycles | Up to 5 cycles |
| on Elecsys® 2010 and cobas e® 411<br>at 20-25 °C | up to 8 hours |
{9}------------------------------------------------
#### d. Detection limits:
Limit of Blank (LoB) of the Elecsys® Anti-HAV assay was determined according to CLSI EP17-A. Limit of Blank determines the highest observed measurement values for samples free of analyte. The LoB was determined as the 95 percentile of measurements of blank samples. The distribution of values for five zero-level human serum has been determined on two Elecsys® 2010 / cobas e 411 Analyzers and on two MODULAR® ANALYTICS E170 / cobas e® 601 Immunoassay Analyzers over 3 days, 2 runs per day. The LoB for Elecsys® 2010 / cobas e 411 Analyzer is 3.03 IU/L and 2.099 IU/L for MODULAR® ANALYTICS E170 / cobas e® 601 Immunoassay Analyzers.
Limit of detection (LoD) of the Elecsys® Anti-HAV assay was determined according to CLSI EP17-A. The LoD was determined as the lowest amount of analyte in a sample that can be detected with 95% probability. The distribution of values for five low-level human serum samples has been determined on two Elecsys® 2010 / cobas e® 411 Analyzers and on two MODULAR® ANALYTICS E170 / cobas e 601 Immunoassay Analyzers over 3 days, 2 runs per day. Samples were measured in one-fold determination in each run. 30 measuring points were collected per instrument.
LOD = LOB + 1.6529 x SD total (of low analyte samples) The LoD for Elecsys® 2010 / cobas e® 411 Analyzer is 5.155 IU/L, and 2.994 IU/L for MODULAR® ANALYTICS E170 / cobas e® 601 Immunoassay Analyzers.
The LOD is set at 6.00 IU/L for both types of analyzers and is reported in the labeling.
Calibrators: Range for the electrochemiluminescence signals for the calibrators:
Negative calibrator (Call): < 20 IU/L (Elecsys® 2010, MODULAR® ANALYTICS E170 and cobas e® analyzers). Positive calibrator (Cal2): ≥ 40 IU/L (Elecsys® 2010, MODULAR® ANALYTICS E170 and cobas e® analyzers)
- e. Analytical specificity:
Cross-reactivity: 177 samples from 15 potentially cross-reactive sub-groups were used for cross-reactivity study on the Elecsys 2010. 174 Samples were found to be nonreactive (negative) in both the Elecsys anti-HAV assay and the predicate AxSym HAVAB 2.0 assay; 3 samples were found to be discordant between the Elecsys anti-HAV assay and the AxSym HAVAB 2.0 assay. The testing results are summarized in the table below:
{10}------------------------------------------------
| | HAVAB 2.0 Assay | | | | | | | | | | |
|----------------------------------------------|----------------------------|----------|-----|----|-----------|----|----|--------------|-----|-------|--|
| | | Reactive | | | Equivocal | | | Non-Reactive | | | |
| Category | Elecsys 2010 HAV 2.1 Assay | | | | | | | | | Total | |
| | RXa | EQb | NRc | RX | EQ | NR | RX | EQ | NR | | |
| Anti-nuclear Antibody (ANA) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 13 | 13 | |
| Cytomegalovirus (anti-CMV) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 13 | 13 | |
| Epstein-Barr Virus (anti-EBV) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 14 | 14 | |
| Elevated IgG | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 13 | 13 | |
| Elevated IgM | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 13 | 12 | |
| Hepatitis B Virus<br>(HBsAg or HBV-DNA) | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 10 | 11 | |
| Hepatitis C Virus (anti-HCV) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 12 | 12 | |
| Human Immunodeficiency Virus<br>(anti-HIV-1) | 0 | 0 | 0 | 0 | 0 | 0 | 2 | 0 | 11 | 13 | |
| Herpes Simplex Virus (anti-HSV) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 10 | 10 | |
| Mumps / Rubeola | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 12 | 12 | |
| Parvovirus B19 Infection | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 11 | 11 | |
| Rheumatoid factor | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 7 | 7 | |
| Rubella (anti-Rubella ) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 10 | 10 | |
| Toxoplasmosis (anti-Toxo) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 13 | 13 | |
| Varicella Zoster (anti-VZV) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 12 | 12 | |
| Total | 0 | 0 | 0 | 0 | 0 | 0 | 3 | 0 | 174 | 177 | |
a Number of Reactive specimens
b Number of Equivocal or Borderline specimens
° Number of Non-Reactive specimens
HAMA (human anti-mouse antibodies) effect was tested by comparing the recovery of 4 human serum samples spiked with different suitable HAMA serum samples versus 4 unspiked aliquots of samples. No HAMA effect was found.
Interference: The impact of endogenous interfering substances on the Elecsys Anti-HAV assay was determined testing native human serum pools on Elecsys® 2010 Immunoassay Analyzer.
The assay is unaffected by icterus (bilirubin < 50 mg/dL), hemolysis (Hb
{11}------------------------------------------------
< 1000 mg/dL), lipemia (Intralipid < 1500 mg/dL), and biotin < 50 ng/mL. Serum pools (high negative, low positive, and high positive) were used to spike with the intereferent. In patients receiving therapy with high biotin doses (i.e. > 5 mg/day), no sample should be taken until at least 8 hours after the last biotin administration. One aliquot of each serum sample was spiked with the interfering substance; another aliquot was spiked with the same volume of isotonic NaCl solution (dilution pool). The interfering pool was then diluted into the dilution pool in 10% increments. The % recovery was determined by dividing the mean value of the measured concentration by the expected concentration. Criterion: Recovery of positive samples within ± 20 % of initial value.
In vitro tests were performed on 18 commonly used pharmaceuticals (Acetylcysteine, Ampicillin-Na, Ascorbic acid. Ca-Dobesilate, Cyclosporine, Cefoxitin, Heparin, Intralipid, Levodopa, Methyldopa, Metronidazole, Phenylbutazone, Tetracycline, Acetylsalicylic acid, Rifampicin, Acetaminophen, Ibuprofen, and Theophylline) and in addition on folic acid. No interference with the assay was found.
- f. Assay cut-off:
The cutoff was set at 20 IU/L according to the WHO recommendation for vaccination. It was established by histogram analysis of the predicate and was confirmed by measuring a total of 736 samples from 2 European centers including hospitalized patients, acute HAV infection patient, blood doors, pregnant women and the other samples that suspected for HAV infection. The 20 IU/L in the Elecsys anti-HAV assay corresponds to 1.0 of S/CO in the predicate.
Non-reactive: < 18 IU/L 18.0 < IU/L < 22.0 IU/L Border: Reactive: > 22.0 IU/L
Samples with a concentration < 18.0 IU/L are considered non-reactive in the Elecsys Anti-HAV test and no further testing is necessary.
Samples with a concentration ≥ 22.0 IU/L are considered reactive in the Elecsys Anti-HAV test.
Samples with a concentration between 18.0 and < 22.0 are considered border (borderline). The sample should be retested in duplicate.
- . If 2 of the 3 results have a concentration < 18.0 IU/L, the result is interpreted as non-reactive and no further testing is necessary.
- If 2 of the 3 results have a concentration between 18.0 and < 22.0 IU/L, the . result is interpreted as borderline. It is recommended that a specimen be drawn in two weeks and retested.
{12}------------------------------------------------
- If 2 of the 3 results have a concentration ≥ 22.0 IU/L, the result is ● interpreted as reactive.
# 2. Comparison studies:
- a. Method comparison with predicate device:
The performance of the Elecsys anti-HAV assay was determined by percent agreement among negative samples and percent agreement among positive samples, against a consensus comparator method, in specific populations. The main predicate (Abbott AxSym HAV AB- 2.0) was used as sole comparator/reference. Detailed information can be found in clinical performance.
- b. Matrix comparison:
The effect on quantitation of analyte in the presence of anticoagulants with the Elecsys® Anti-HAV Immunoassay was determined on Elecsys 2010 Immunoassay Analyzer by comparing values obtained from native samples (single donors) drawn into Serum, K2-EDTA-Plasma and Serum-Gel Separation primary tubes. The following tables summarize the results for the comparison between serum and plasma matrices.
| Plasma matrix | Number of positive specimens showing recovery to<br>serum within various ranges | | |
|---------------|---------------------------------------------------------------------------------|--------|-------|
| | < 10% | 10-15% | > 15% |
| K2-EDTA | 24 | 1 | 0 |
| Serum Gel | 23 | 2 | 0 |
| Plasma matrix | Number of borderline specimens showing recovery<br>to serum within various ranges | | |
|---------------|-----------------------------------------------------------------------------------|--------|-------|
| | < 10% | 10-15% | > 15% |
| K2-EDTA | 3 | 0 | 0 |
| Serum Gel | 3 | 0 | 0 |
| Plasma matrix | Number of negative specimens showing recovery<br>to serum within various ranges | | |
|---------------|---------------------------------------------------------------------------------|---------------|------------|
| | < 1.0 IU/L | 1.0 -2.0 IU/L | > 2.0 IU/L |
| K2-EDTA | 29 | 1 | 0 |
| Serum Gel | 30 | 0 | 0 |
{13}------------------------------------------------
## 3. Clinical studies:
- a. Clinical Sensitivity: N/A
- b. Clinical specificity: N/A
- c. Other clinical supportive data (when a. and b. are not applicable):
# Clinical Performance
### Clinical Study Cohorts:
A multi-center study was conducted in the U.S. to characterize the performance of the Elecsys Anti-HAV Immunoassay. All subjects were tested with the Elecsys Anti-HAV assay on the Elecsys 2010 analyzer and with an FDAcleared reference method in strict accordance with the manufacturer's package insert instructions.
A total of 1050 samples were obtained from multiple specimen sources, representing subjects for whom routine hepatitis A testing had been ordered. hospitalized patients, subjects at increased risk for hepatitis, subjects with signs and symptoms of hepatitis, subjects characterized with acute hepatitis A, and subjects below the age of 21 years (pediatric/adolescents). All samples (prospective and retrospective) are stored frozen before shipment to Roche and to the respective sites for testing. Among the 1050 samples, 487 were (46.38%) were from males and 563 (53.62%) were from females. 0.38% were from American Indian/Alaska Native, 2.76% were Asian, 15.90% were African American /Black, 65.14% were Caucasian/White, 14.95% unknown and 0.86% others.
Among all specimens tested, one specimen subjected to routine Hepatitis A testing was concordantly equivocal, and was excluded from the calculations.
The positive and negative percent agreement for prospectively collect samples is 98.13% (95.96% to 99.31%) and 97.37% (95.06% to 98.79%), respectively.
The positive and negative percent agreement for retrospectively collect samples is 98.11% (94.59% to 99.61%) and 97.37% (94.36% to 99.03%), respectively.
The positive percent agreement and the negative percent agreement results for the different clinical population are presented in the following table:
{14}------------------------------------------------
| Elecsys<br>2010 Result | Predicate Result | | | | | | | | | | | |
|------------------------|------------------|-----------|-----------|----------------|----|----|-----------------|----|----|-----------------|----|-----|
| | Site 1 BW | Site 2 WU | Site 3 JH | All Sites | | | | | | | | |
| | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR |
| Reactive | 60 | 0 | 2 | 0 | 0 | 0 | 16 | 0 | 0 | 76 | 0 | 2 |
| Equivocal | 1 | 0 | 2 | 0 | 0 | 0 | 1 | 0 | 0 | 2 | 0 | 2 |
| Negative | 0 | 0 | 97 | 0 | 0 | 0 | 0 | 0 | 17 | 0 | 0 | 114 |
| Total | 61 | 0 | 101 | 0 | 0 | 0 | 17 | 0 | 17 | 78 | 0 | 118 |
| PPA | 98.36 (60/61) | | | 0.00 (0/0) | | | 94.12 (16/17) | | | 97.44 (76/78) | | |
| 95% CI | 91.20 to 99.96 | | | 0.00 to 100.00 | | | 71.31 to 99.85 | | | 91.04 to 99.69 | | |
| NPA | 96.04 (97/101) | | | 0.00 (0/0) | | | 100.00 (17/17) | | | 96.61 (114/118) | | |
| 95% CI | 90.17 to 98.91 | | | 0.00 to 100.00 | | | 80.49 to 100.00 | | | 91.55 to 99.07 | | |
Percent Agreement for Elecsys and Predicate anti-HAV Assay Results from Symptomatic Individuals
Percent Agreement for Elecsys and Predicate anti-HAV Assay Results from Specimens Subjected to Routine Hepatitis A Testing
| | Predicate Result | | | | | | | | | | | |
|------------------------|------------------|---------------|----|----------------|----|----|-----------------|----|----|-----------------|----|----|
| Elecsys<br>2010 Result | Site 1 BW | | | Site 2 WU | | | Site 3 JH | | | All Sites | | |
| | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR |
| Reactive | 90 | 0 | 2 | 0 | 0 | 0 | 36 | 0 | 1 | 126 | 0 | 3 |
| Equivocal | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 1 | 1 | 0 | 1 | 2 |
| Negative | 2 | 0 | 54 | 0 | 0 | 0 | 0 | 0 | 21 | 2 | 0 | 75 |
| Total | 92 | 0 | 57 | 0 | 0 | 0 | 36 | 1 | 23 | 128 | 1 | 80 |
| PPA | | 97.83 (90/92) | | 0.00 (0/0) | | | 100.00 (36/36) | | | 98.44 (126/128) | | |
| 95% CI | 92.37 to 99.74 | | | 0.00 to 100.00 | | | 90.26 to 100.00 | | | 94.47 to 99.81 | | |
| NPA | | 94.74 (54/57) | | 0.00 (0/0) | | | 91.30 (21/23) | | | 93.57 (75/80) | | |
| Percent Agreement for Elecsys and Predicate anti-HAV Assay Results from | |
|-------------------------------------------------------------------------|--|
| Hospitalized Patients (Retrospective and Prospective) | |
| | | Predicate Result | | | | | | | | | | | |
|------------------------|--|------------------|----|-----------|----|-----------|----|-----------|----|----|----|----|-----|
| | | Site 1 BW | | Site 2 WU | | Site 3 JH | | All Sites | | | | | |
| Elecsys<br>2010 Result | | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR |
| Reactive | | 42 | 0 | 0 | 0 | 0 | 0 | 28 | 0 | 0 | 70 | 0 | 0 |
| Equivocal | | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 1 | 0 | 0 |
| Negative | | 0 | 0 | 90 | 0 | 0 | 0 | 1 | 0 | 51 | 1 | 0 | 141 |
{15}------------------------------------------------
| Total | 42 | 0 | 90 | 0 | 0 | 0 | 30 | 0 | 51 | 72 | 0 | 141 |
|--------|-----------------|---|----|----------------|----------------|---|----|-----------------|----|------------------|---|-----|
| PPA | 100.00 (42/42) | | | 0.00 (0/0) | | | | 93.33 (28/30) | | 97.22 (70/72) | | |
| 95% CI | 91.59 to 100.00 | | | | 0.00 to 100.00 | | | 77.93 to 99.18 | | 90.32 to 99.66 | | |
| NPA | 100.00 (90/90) | | | | 0.00 (0/0) | | | 100.00 (51/51) | | 100.00 (141/141) | | |
| 95% CI | 95.98 to 100.00 | | | 0.00 to 100.00 | | | | 93.02 to 100.00 | | 97.42 to 100.00 | | |
# Percent Agreement for Elecsys and Predicate anti-HAV Assay Results from Subjects Characterized as Acute Hepatitis A (Retrospective and Prospective)
| | Predicate Result | | | | | | | | | | | |
|---------------------|------------------|----|----|----------------|----|----|-----------------|----|----|------------------|----|----|
| | Site 1 BW | | | Site 2 WU | | | Site 3 JH | | | All Sites | | |
| Elecsys 2010 Result | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR |
| Reactive | 81 | 0 | 0 | 0 | 0 | 0 | 50 | 0 | 0 | 131 | 0 | 0 |
| Equivocal | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| Negative | 0 | 0 | 3 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 3 |
| Total | 81 | 0 | 3 | 0 | 0 | 0 | 50 | 0 | 0 | 131 | 0 | 3 |
| PPA | 100.00 (81/81) | | | 0.00 (0/0) | | | 100.00 (50/50) | | | 100.00 (131/131) | | |
| 95% CI | 95.55 to 100.00 | | | 0.00 to 100.00 | | | 92.89 to 100.00 | | | 97.22 to 100.00 | | |
| NPA | 100.00 (3/3) | | | 0.00 (0/0) | | | 0.00 (0/0) | | | 100.00 (3/3) | | |
| 95% CI | 29.24 to 100.00 | | | 0.00 to 100.00 | | | 0.00 to 100.00 | | | 29.24 to 100.00 | | |
# Percent Agreement for Elecsys and Predicate anti-HAV Assay Results from Pediatric and Adolescent Subjects
| | Predicate Result | | | | | | | | | | | |
|------------------------|------------------|----|----|----------------|----|----|-----------------|----|----|-----------------|----|----|
| | Site 1 BW | | | Site 2 WU | | | Site 3 JH | | | All Sites | | |
| Elecsys<br>2010 Result | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR |
| Reactive | 0 | 0 | 0 | 0 | 0 | 0 | 19 | 0 | 2 | 19 | 0 | 2 |
| Equivocal | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| Negative | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 65 | 0 | 0 | 65 |
| Total | 0 | 0 | 0 | 0 | 0 | 0 | 19 | 0 | 67 | 19 | 0 | 67 |
| PPA | 0.00 (0/0) | | | 0.00 (0/0) | | | 100.00 (19/19) | | | 100.00 (19/19) | | |
| 95% CI | 0.00 to 100.00 | | | 0.00 to 100.00 | | | 82.35 to 100.00 | | | 82.35 to 100.00 | | |
| NPA | 0.00 (0/0) | | | 0.00 (0/0) | | | 97.01 (65/67) | | | 97.01 (65/67) | | |
| 95% CI | 0.00 to 100.00 | | | 0.00 to 100.00 | | | 89.63 to 99.64 | | | 89.63 to 99.64 | | |
{16}------------------------------------------------
| | Predicate Result | | | | | | | | | | | |
|------------------------|------------------|----|----|----------------|----|----|-----------------|----|-----|-----------------|----|-----|
| | Site 1 BW | | | Site 2 WU | | | Site 3 JH | | | All Sites | | |
| Elecsys<br>2010 Result | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR |
| Reactive | 10 | 0 | 1 | 0 | 0 | 0 | 38 | 0 | 2 | 48 | 0 | 3 |
| Equivocal | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 1 |
| Negative | 1 | 0 | 5 | 0 | 0 | 0 | 2 | 0 | 152 | 3 | 0 | 157 |
| Total | 11 | 0 | 6 | 0 | 0 | 0 | 40 | 0 | 155 | 51 | 0 | 161 |
| PPA | 90.91 (10/11) | | | 0.00 (0/0) | | | 95.00 (38/40) | | | 94.12 (48/51) | | |
| 95% CI | 58.72 to 99.77 | | | 0.00 to 100.00 | | | 83.08 to 99.39 | | | 83.76 to 98.77 | | |
| NPA | 83.33 (5/6) | | | 0.00 (0/0) | | | 98.06 (152/155) | | | 97.52 (157/161) | | |
| 95% CI | 35.88 to 99.58 | | | 0.00 to 100.00 | | | 94.45 to 99.60 | | | 93.76 to 99.32 | | |
Agreement for Elecsys and Predicate anti-HAV Assay Results from Subjects at Increased Risk for Hepatitis
RX - Reactive, positive, EQ - equivocal, borderline, NR - non-reactive, negative, PPA - positive percent agreement, NPA negative percent agreement, 95% CI - 95% Exact confidence interval
| Cohort | Positive Percent Agreement | Negative Percent Agreement | | |
|------------------------------|----------------------------|----------------------------|------------------|--------------------------|
| | PPA (x/n) | 95 % confidence interval | NPA (x/n) | 95 % confidence interval |
| Routine HAV Testing | 98.44 (126/128) | 94.47 to 99.81 | 93.75 (75/80) | 86.01 to 97.94 |
| Hospitalized | 97.22 (70/72) | 90.32 to 99.66 | 100.00 (141/141) | 97.42 to 100.00 |
| Signs and Symptoms | 97.44 (76/78) | 91.04 to 99.69 | 96.61 (114/118) | 91.55 to 99.07 |
| Increased Risk for Hepatitis | 94.12 (48/51) | 83.76 to 98.77 | 97.52 (157/161) | 93.76 to 99.32 |
| Characterized Acute HAV | 100.00 (131/131) | 97.22 to 100.00 | 100.00 (3/3) | 29.24 to 100.00 |
| Pediatric/Adolescent | 100.00 (19/19) | 82.35 to 100.0 | 97.01 (65/67) | 89.63 to 99.64 |
| Overall | 98.12 (470/479) | 96.46 to 99.14 | 97.37 (555/570) | 95.70 to 98.52 |
### Summary of the Percent Agreements for the Various Specimen Cohorts
### HAV Vaccination:
Fifty-four specimens that were collected both pre- and post-HAV vaccination was evaluated by the Elecsys anti-HAV assay and the predicate. The post-vaccination specimens were obtained at least four weeks but not more than 10 weeks after the completion of the vaccine regimen. Three HAV vaccines which are currently licensed in the US were used in this study: VAQTA (16), HAVRIX (20) and TWINRIX (18). Two vaccination studies are represented: one conducted in the Eastern region of the US and the other in Penzberg Germany. There was no antibody response observed in pre-vaccination specimens with the exception of two
{17}------------------------------------------------
specimens which were just above the equivocal/borderline (- 23.23 IU/L) and just reactive (30.34 IU/L), respectively.
| | anti-HAV Assay Results | | | | | | | | | | | |
|-------------------|------------------------|----|----|----------------|----|----|-----------------|----|----|----------------|----|----|
| | Predicate Result | | | | | | | | | | | |
| Elecsys<br>Result | HAVRIX | | | TWINRIX | | | VAQTA | | | All Vaccines | | |
| | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR |
| Reactive | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 1 |
| Equivocal | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| Non-<br>Reactive | 0 | 0 | 20 | 0 | 0 | 17 | 0 | 0 | 16 | 0 | 0 | 53 |
| Total | 0 | 0 | 20 | 0 | 0 | 18 | 0 | 0 | 16 | 0 | 0 | 54 |
| PPA | 0.00 (0/0) | | | 0.00 (0/0) | | | 0.00 (0/0) | | | 0.00 (0/0) | | |
| 95% CI* | 0.00 to 100.00 | | | 0.00 to 100.00 | | | 0.00 to 100.00 | | | 0.00 to 100.00 | | |
| NPA | 100.00 (20/20) | | | 94.44 (17/18) | | | 100.00 (16/16) | | | 98.15 (53/54) | | |
| 95% CI* | 83.16 to 100.00 | | | 72.71 to 99.86 | | | 79.41 to 100.00 | | | 90.11 to 99.95 | | |
| Percent Agreement for Elecsys and Predicate anti-HAV Assays Results from |
|--------------------------------------------------------------------------|
| Pre-HAV Vaccination Specimens |
## Percent Agreement for Elecsys and Predicate anti-HAV Assays Results from Post-HAV Vaccination Specimens
| anti-HAV Assay Results | | | | | | | | | | | | |
|------------------------|------------------|----|----|-----------------|----|----|-----------------|----|----|-----------------|----|----|
| | Predicate Result | | | | | | | | | | | |
| Elecsys<br>Result | HAVRIX | | | TWINRIX | | | VAQTA | | | All Vaccines | | |
| | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR | RX | EQ | NR |
| Reactive | 20 | 0 | 0 | 17 | 0 | 1 | 16 | 0 | 0 | 53 | 0 | 1 |
| Equivocal | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| Non-<br>Reactive | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| Total | 20 | 0 | 0 | 17 | 0 | 1 | 16 | 0 | 0 | 53 | 0 | 1 |
| PPA | 100.00 (20/20) | | | 100.00 (17/17) | | | 100.00 (16/16) | | | 100.00 (53/53) | | |
| 95% CI* | 83.16 to 100.00 | | | 80.49 to 100.00 | | | 79.41 to 100.00 | | | 93.28 to 100.00 | | |
| NPA | 0.00 (0/0) | | | 0.00 (0/1) | | | 0.00 (0/0) | | | 0.00 (0/1) | | |
| 95% CI* | 0.00 to 100.00 | | | 0.00 to 97.50 | | | 0.00 to 100.00 | | | 0.00 to 97.50 | | |
RX - Reactive, positive, EQ - equivocal, borderline, NR - non-reactive, negative, PPA - positive percent agreement, NPA negative percent agreement, 95% CI - 95% Exact confidence interval
## Prevalence Studies:
{18}------------------------------------------------
The Elecsys Anti-HAV assay was used to evaluate the prevalence of HAV total antibodies in an apparently healthy population (normal, healthy individuals without symptoms). The prospective study population for the Elecsys Anti-HAV assay consisted of 602 patients. Of these 602 patients, 300 patients were from the high prevalence region, Western states of the U.S. (New Mexico) and 302 patients were from the low risk region Eastern states of the U.S. (Indiana). The prospective study population was 208 (34.6 %) males and 394 (65.4 %) females (total n = 602) including 493 (81.9 %) Caucasians, 32 (5.3 %) African Americans, 6 (1.0 %) Asians, 69 (11.5 %) American Indians and 2 (0.3 %) Others. The results of prevalence population are summarized according to age groups in decades, gender, geographical area and the number of reactive, non-reactive and equivocal results.
Prevalence rate for reactive anti-HAV assay in specimens collected in a low prevalence region, Eastern states of the US (Indiana), was 35.10 %. Prevalence rate for reactive anti-HAV assay in specimens collected in a high prevalence region, Western states of the US (New Mexico), was 37.33%. The results of prevalence study are summarized according to age groups and gender.
| Expected results for the Elecsys Anti-HAV assay in subjects from low prevalence | |
|---------------------------------------------------------------------------------|--|
| areas for Hepatitis A | |
| Age<br>range<br>(years) | Gender | Elecsys Anti-HAV result | | | | | | |
|-------------------------|--------|-------------------------|---------|-----------|---------|--------------|---------|-------|
| | | Reactive | | Equivocal | | Non-reactive | | |
| | | N | Percent | N | Percent | N | Percent | Total |
| 11 - 20 | Female | 0 | 0.0 | 0 | 0.0 | 1 | 100.0 | 1 |
| 11 - 20 | Male | 0 | 0.0 | 0 | 0.0 | 1 | 100.0 | 1 |
| 21-30 | Female | 1 | 14.3 | 0 | 0.0 | 6 | 85.7 | 7 |
| 21-30 | Male | 0 | 0.0 | 0 | 0.0 | 6 | 100.0 | 6 |
| 31-40 | Female | 2 | 9.5 | 1 | 4.8 | 18 | 85.7 | 21 |
| 31-40 | Male | 0 | 0.0 | 0 | 0.0 | 2 | 100.0 | 2 |
| 41-50 | Female | 3 | 13.6 | 1 | 4.6 | 18 | 81.8 | 22 |
| 41-50 | Male | 1 | 7.7 | 0 | 0.0 | 12 | 92.3 | 13 |
| 51-60 | Female | 7 | 16.7 | 1 | 2.4 | 34 | 81.0 | 42 |
| 51-60 | Male | 9…
