K Number
K063318

Validate with FDA (Live)

Date Cleared
2007-05-03

(182 days)

Product Code
Regulation Number
866.3310
Age Range
All
Reference & Predicate Devices
N/A
Predicate For
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticPediatricDiagnosticis PCCP AuthorizedThirdpartyExpeditedreview
Intended Use

The MONOLISA™ Anti-HAV EIA is an in vitro enzyme immunoassay kit intended for use in the qualitative detection of total antibodies (IgG and IgM) to Hepatitis A Virus (anti-HAV) in human (adult and pediatric) serum or plasma (EDTA, Heparin, Citrate, ACD). This kit can be used as an aid in the diagnosis of acute or past Hepatitis A Virus (HAV) infection or as an aid in the identification of HAVsusceptible individuals for vaccination. However, any diagnosis should take into consideration the patient's clinical history and symptoms, as well as serological data.

Assay performance characteristics have not been established for immunocompromised or immunosuppressed patients, and cord blood or neonatal specimens.

WARNING : This assay is not intended for screening blood or solid or soft tissue donors.

Device Description

The MONOLISA™ Anti-HAV EIA is an enzyme immunoassay (competitive assay format) for the detection of total antibodies to Hepatitis A virus. In the assay procedure, patient specimens, a calibrator and controls are incubated with HAV antigen in microwells that have been coated with mouse monoclonal anti-Hepatitis A antibodies to HAV present in a specimen or control will complex with the HAV antigen reagent and with antibodies coated on the microwells. Excess sample and HAV Viral antigen reagent are removed by a wash step. The conjugate (containing horseradish peroxidase-labeled mouse monoclonal antibody to HAV) is subsequently added to the microwells and incubated. The conjuqate binds to the HAV antigen bound to the microwell in the absence of antibodies to HAV from the specimen. Excess conjugate is removed by a wash step, and a TMB Chromogen / Substrate solution is added to the microwells and allowed to incubate. If a sample does not contain anti-HAV antibodies, the bound enzyme (HRP) causes the colorless tetramethylbenzidine (TMB) in the Chromogen solution to change to blue. The blue color turns yellow after the addition of a Stopping Solution. If a sample contains anti-HAV antibodies, the Chromogen / Substrate Solution in the well remains colorless during the substrate incubation, and after the addition of the Stopping Solution. The color intensity is measured spectrophotometrically.

Absorbance value readings for patient specimens are compared to the Cutoff value determined by the mean of the Calibrator absorbance values.

AI/ML Overview

Here's a breakdown of the acceptance criteria and study information for the MONOLISA™ Anti-HAV EIA, based on the provided document:

Acceptance Criteria and Reported Device Performance

CriteriaAcceptance Criteria (Implied by reported performance vs. comparative assay)Reported Device Performance (MONOLISA™ Anti-HAV EIA)
Overall Positive Percent Agreement (PPA) with comparative assayNot explicitly stated, but generally expected to be very high (e.g., >95%)99.5% (931/936) with 95% CI (98.8% - 99.8%)
Overall Negative Percent Agreement (NPA) with comparative assayNot explicitly stated, but generally expected to be very high (e.g., >95%)96.2% (378/393) with 95% CI (93.8% - 97.9%)
PPA for US populationNot explicitly stated98.8% (237/240) with 95% CI (96.4% - 99.7%)
NPA for US populationNot explicitly stated92.6% (151/163) with 95% CI (87.5% - 96.1%)
PPA for European populationNot explicitly stated99.7% (610/612) with 95% CI (98.8% - 99.9%)
NPA for European populationNot explicitly stated98.7% (227/230) with 95% CI (96.2% - 99.7%)
PPA for Acute HAV InfectionNot explicitly stated100% (84/84) with 95% CI (96.5% - 100%)
PPA for Pediatric SubjectsNot explicitly stated100% (29/29) with 95% CI (90.2% - 100%)
NPA for Pediatric SubjectsNot explicitly stated96.8% (30/31) with 95% CI (83.3% - 99.9%)
Seroconversion Panel Sensitivity (compared to comparative assay)Equivalent or more sensitiveEquivalent to or more sensitive in 6/6 panels.
Device Precision (Within-run, Between-run, Between-day CV)CVs typically expected to be low (e.g., <15-20%)Refer to tables 17, 18, 19, 20 for detailed CVs. Generally: <12% for most individual categories, some total CVs up to 28% for certain panel members at specific sites in reproducibility studies.
Device Reproducibility (Across sites, lots, operators CV)CVs typically expected to be low (e.g., <15-20%)Refer to tables 18, 19, 20 for detailed CVs. Generally: <15% for most individual categories for controls/low reactive samples, but some total CVs as high as ~24% for higher reactive panel members.
Cross-ReactivityLow to no clinically significant cross-reactivity with common interfering conditions/viruses.7 samples were discrepant (4 reactive with MONOLISA™ Anti-HAV EIA but non-reactive with comparative assay; 3 non-reactive with MONOLISA™ Anti-HAV EIA but reactive with comparative assay) out of 255 tested across 16 clinical conditions. Specific rates per condition are listed in Table 16.

Study Information

  1. Sample Size and Data Provenance (Test Set):

    • Overall Test Set: 1327 samples (404 US, 928 European) for clinical performance, plus additional samples for acute HAV infection (84), pediatrics (60 additional to acute HAV), vaccinated subjects (62 pre/post-vaccination from 38 individuals, plus 14 purchased post-vaccination), seroconversion panels (6 panels), and cross-reactivity (255 specimens).
    • Clinical Performance (US): 404 specimens (174 with signs/symptoms of Hepatitis, 230 high-risk).
      • Country of Origin: US (Los Angeles, Santa Ana, CA, and Miami, FL).
      • Retrospective/Prospective: Both prospective and retrospective.
    • Clinical Performance (Europe): 928 specimens (252 signs/symptoms of Hepatitis, 62 high-risk, 345 asymptomatic hospitalized, 34 healthcare workers, 151 recovered HAV infection).
      • Country of Origin: Europe (France and Italy).
      • Retrospective/Prospective: Both prospective and retrospective.
    • Acute HAV Infection: 84 retrospective samples (European population, adult and pediatric).
    • Pediatric Subjects: 60 samples from US and Europe (in addition to the 39 from acute HAV infection).
    • High Risk Individuals (Expected Values):
      • US: 230 subjects (Los Angeles, CA; Santa Ana, CA; Miami, FL).
      • Europe: 62 subjects.
    • Healthy Individuals (Expected Values):
      • Mid-west US: 280 subjects (St. Louis, Missouri).
      • Western US: 245 subjects (California and Washington).
      • Europe: 285 subjects (Parma, Italy).
    • Vaccinated Subjects: 62 pre- and post-vaccination samples from 38 individuals (US and Europe), plus 14 purchased post-vaccination samples (US).
    • Seroconversion Panels: 6 commercially available panels.
    • Cross-Reactivity Study: 255 specimens from 16 clinical conditions.
  2. Number of Experts and Qualifications for Ground Truth (Test Set):

    • The document mentions comparison against a "comparative assay" (predicate device, DiaSorin ETI-AB-HAVK PLUS, PMA Number: P890019) and "FDA approved methods" to confirm disease state for cross-reactivity.
    • It does not explicitly state the number of experts or their qualifications for establishing the ground truth for the test set. For clinical assays like this, it is common for the ground truth to be established by the results of a previously approved, well-established method (the comparative assay) or by a composite reference standard that might involve expert clinical diagnosis and other laboratory tests, but this detail is not provided.
  3. Adjudication Method (Test Set):

    • The primary method was comparison against a "comparative assay" (predicate device).
    • For instances where the MONOLISA™ Anti-HAV EIA produced a borderline result or where the comparative assay produced a borderline result, specific rules were applied for agreement calculations (e.g., a borderline comparative assay result reactive with MONOLISA™ was considered a false positive for MONOLISA™).
    • There is no mention of a traditional expert adjudication panel (e.g., 2+1, 3+1).
  4. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study:

    • No, a MRMC comparative effectiveness study was not done. This is an automated in vitro diagnostic device, not an imaging device that would typically involve human readers. The comparison is between the new device and a predicate device.
  5. Standalone Performance Study:

    • Yes, Standalone performance was done. The entire clinical and analytical performance evaluation describes the algorithm's (device's) performance in isolation against a ground truth (the comparative assay/FDA approved methods), without human interpretation in the loop other than performing the assay and reading the spectrophotometric results.
  6. Type of Ground Truth Used:

    • For clinical performance studies, the ground truth was predominantly established by the results of a "comparative assay" (the legally marketed equivalent device, DiaSorin ETI-AB-HAVK PLUS).
    • For the cross-reactivity study, "FDA approved methods" were used to confirm the disease state of each specimen.
    • For acute HAV infection, samples were "from subjects with a medical history and laboratory results indicative of acute Hepatitis A."
    • For vaccinated subjects, the ground truth was pre/post-vaccination status as determined by clinical trial enrollment and/or other laboratory methods in addition to the comparative assay.
  7. Sample Size for the Training Set:

    • The document describes performance studies (validation). It does not explicitly mention a separate "training set" sample size for machine learning or AI models. Given that it's an enzyme immunoassay kit, it's a biochemical assay rather than a predictive algorithm that undergoes a training phase in the typical AI sense. The development of such a kit would involve internal optimization and calibration (which could be considered analogous to training) but not usually with a distinctly defined 'training set' of patient samples as seen in AI/ML validation studies.
  8. How Ground Truth for the Training Set Was Established:

    • As noted above, a distinct "training set" for an AI/ML model isn't described. For the biochemical assay, internal controls, calibrators, and development batches would have been used during the product development phase. The "ground truth" for these would be based on known concentrations or presence/absence of analytes, established through established laboratory methods and reference materials, but these details are not provided in the 510(k) summary (which focuses on validation).

{0}------------------------------------------------

Image /page/0/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and distributes products for the life science research and clinical diagnostics markets. The logo consists of the text "BIO+RAD" in a bold, sans-serif font. The text is white and is set against a black, rounded rectangle.

MONOLISA™ Anti-HAV EIA 510(k) SUMMARY

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

510(k) Number:K 063318
510(k) Summary Report Date:April, 2007
MANUFACTURER INFORMATION
MANUFACTURER ADDRESS:Bio-Rad3, Boulevard Raymond Poincaré92430 Marnes-la-Coquette, France
TELEPHONE :00 33 1 47 95 60 00
ESTABLISHMENT REG. NUMBER :8023060
OWNER/OPERATOR :Bio-Rad3, Boulevard Raymond Poincaré92430 Marnes-la-Coquette, France
OWNER/ OPERATOR NUMBER :8023061
OFFICIAL CORRESPONDENTADDRESS :Bio-Rad3, Boulevard Raymond Poincaré92430 Marnes-la-Coquette, France
TELEPHONE:00 33 1 47 95 60 00
OFFICIAL CORRESPONDENT :Mrs. Sylvie Confida
TELEPHONE :FAX :00 33 1 47 95 61 3800 33 1 47 95 62 42
CLASSIFICATION INFORMATION
COMMON NAME:Total Antibody to Hepatitis A Virus
PRODUCT TRADE NAME:MONOLISA™ Anti-HAV EIA
DEVICE CLASS:Class II LOL

·

{1}------------------------------------------------

Image /page/1/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and distributes products for the life science research and clinical diagnostics markets. The logo is in white text on a black background. The text is bold and sans-serif.

CLASSIFICATION PANEL :

Immunology and Microbiology Devices

REGULATION NUMBER : 21 CFR 866.3310

LEGALLY MARKETED EQUIVALENT (SE) DEVICE

DiaSorin ETI-AB-HAVK PLUS PMA Number : P890019 Decision Date : 12/12/2005

DEVICE DESCRIPTION

The MONOLISA™ Anti-HAV EIA is an enzyme immunoassay (competitive assay format) for the detection of total antibodies to Hepatitis A virus. In the assay procedure, patient specimens, a calibrator and controls are incubated with HAV antigen in microwells that have been coated with mouse monoclonal anti-Hepatitis A antibodies to HAV present in a specimen or control will complex with the HAV antigen reagent and with antibodies coated on the microwells. Excess sample and HAV Viral antigen reagent are removed by a wash step. The conjugate (containing horseradish peroxidase-labeled mouse monoclonal antibody to HAV) is subsequently added to the microwells and incubated. The conjuqate binds to the HAV antigen bound to the microwell in the absence of antibodies to HAV from the specimen. Excess conjugate is removed by a wash step, and a TMB Chromogen / Substrate solution is added to the microwells and allowed to incubate. If a sample does not contain anti-HAV antibodies, the bound enzyme (HRP) causes the colorless tetramethylbenzidine (TMB) in the Chromogen solution to change to blue. The blue color turns yellow after the addition of a Stopping Solution. If a sample contains anti-HAV antibodies, the Chromogen / Substrate Solution in the well remains colorless during the substrate incubation, and after the addition of the Stopping Solution. The color intensity is measured spectrophotometrically.

Absorbance value readings for patient specimens are compared to the Cutoff value determined by the mean of the Calibrator absorbance values.

{2}------------------------------------------------

Image /page/2/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that develops, manufactures, and markets a broad range of products and solutions for the life science research and clinical diagnostic markets. The logo is white text on a black background. The text is in a bold, sans-serif font.

KIT COMPONENTS

ComponentDescription
R1 Microwell Strip PlatesTwo (2) x 12 strips of 8 wells coated with monoclonal anti-HAVantibodies.
R2 Wash SolutionConcentrate (30x)One (1) 120 mL bottle, Tris-buffer containing NaCl and Tween 20.
C0 Negative ControlOne (1) 1.5 mL vial, containing human serum, negative for total anti-HAV antibodies, HBs antigen, anti-HCV antibodies and anti-HIV-1/HIV-2 antibodies. Preservatives : Sodium azide (< 0.1%) and Proclin™300 (0.25%).
C1 Positive ControlOne (1) 1.5 mL vial, containing human serum, positive for anti-HAVantibodies and negative for HBs antigen, anti-HCV antibodies andanti-HIV-1/HIV-2 antibodies, diluted in human serum pool negative foranti-HAV antibodies. Preservatives : Sodium azide (< 0.1%) andProclin™ 300 (0.25%).
C2 CalibratorTwo (2) x 1.5 mL vials, containing human serum, positive for anti-HAVantibodies, and negative for HBs Antigen, anti-HCV antibodies andanti-HIV-1/HIV-2 antibodies, diluted in human serum pool negative foranti-HAV antibodies. Preservatives : Sodium azide (< 0.1%) andProclin™ 300 (0.25%).
R6 HAV Viral AntigenTwo (2) x 14 mL bottles, inactivated HAV virus in Tris buffer containingproteins and sample indicator dye. Preservative : Proclin™ 300 (0.1%).
R7 ConjugateTwo (2) x 14 mL bottles, conjugate (Peroxidase labeled mousemonoclonal antibody to HAV) in Tris buffer containing proteins,detergent and sample indicator dye. Preservative : Proclin™ 300(0.1%).
R8 Substrate BufferOne (1) 120 mL bottle, containing Hydogen Peroxide, citric acid /sodium acetate buffer and Dimethylsulfoxide (DMSO).
R9 Chromogen (11x)One (1) 12 mL bottle, containing Tetramethylbenzidine (TMB).
R10 Stopping SolutionOne (1) 120 mL bottle, containing 1 N H2SO4.
Plate sealersEight (8) clear plastic sealers.

{3}------------------------------------------------

Image /page/3/Picture/0 description: The image shows the logo for Bio-Rad. The logo is black and white and features the text "BIO-RAD" in a bold, sans-serif font. The text is enclosed in a rounded rectangle. The logo is simple and recognizable.

INTENDED USE

The MONOLISA™ anti-HAV EIA is an in vitro enzyme immunoassay kit intended for use in the qualitative detection of total antibodies (anti-HAV IgG and IgM) to Hepatitis A virus (anti-HAV) in human (adult and pediatric) serum or plasma (EDTA, Heparin, Citrate, ACD.

This assay is not intended for screening blood or solid or soft tissue donors.

INDICATIONS FOR USE

The MONOLISA™ anti-HAV EIA is indicated for use as an aid in the diagnosis of acute or past Hepatitis A Virus (HAV) infection or as an aid in the identification of HAV-susceptible individuals for vaccination. However, any diagnosis should take into consideration the patient's clinical history and symptoms, as well as serological data.

Assay performance characteristics have not been established for immunocompromised or immunosuppressed patients, and core blood or neonatal specimens.

TECHNOLOGICAL CHARACTERISTICS

The following tables summarize similarities and differences between the MONOLISA™ Anti-HAV EIA kit and the predicate device ETI-AB-HAVK PLUS.

Similarities inMONOLISAT™ Anti-HAVETI-AB-HAVK PLUS
Components / MaterialsEIACatalog# P001926
Catalog# 72496
Solid PhaseMicroplate wells coated withmouse Monoclonal anti-HAVantibodies.Microplate wells coated withmouse Monoclonal anti-HAVantibodies.
ConjugatePeroxidase-labeled mousemonoclonal antibody to HAV.Peroxidase-labeled mousemonoclonal antibody to HAV.
Negative ControlHuman serum, negative fortotal anti-HAV antibodies.Human serum/plasma,negative for total anti-HAVantibodies.
CalibratorHuman serum, positive foranti-HAV antibodies, dilutedin human serum poolnegative for anti-HAVantibodies.Human serum/plasma,containing anti-HAVantibodies.
Positive ControlHuman serum, positive foranti-HAV antibodies, dilutedin human serum poolnegative for anti-HAVantibodies.Human serum/plasma,reactive for anti-HAVantibodies.
ChromogenTetramethylbenzidine (TMB)Tetramethylbenzidine (TMB)

Table 1: Similarities between kit components and materials

{4}------------------------------------------------

Image /page/4/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and distributes products for the life science research and clinical diagnostics markets. The logo is white text on a black background. The text is in a bold, sans-serif font.

SubstrateHydrogen PeroxideHydrogen Peroxide
Washing SolutionConcentrated bufferedsolution with Tween 20.Concentrated bufferedsolution with detergents.

Table 2: Differences between kit components and materials

Differences inComponents / MaterialsMONOLISA™ Anti-HAVEIACatalog# 72496ETI-AB-HAVK PLUSCatalog# P001926
ConjugateReady-to-use.To be diluted.
Incubation bufferNABuffer, containing proteinstabilizers and an inert bluedye.
Viral Antigen / NeutralizingSolutionTris-buffer, containinginactivated HAV-virus,proteins and sampleindicator dye.Buffer, containing HAV,human serum/plasma andprotein stabilizers.
Stopping Solution$1N H_2SO_4$ .$0.4N H_2SO_4$ .

{5}------------------------------------------------

Image /page/5/Picture/0 description: The image shows the logo for Bio-Rad. The logo is white text on a black rounded rectangle. The text reads "BIO-RAD" in all capital letters.

Similarities in Functionand UseMONOLISA™ Anti-HAVEIACatalog# 72496ETI-AB-HAVK PLUSCatalog# P001926
Test MethodEIA (competitive assay format)EIA (competitive test)
Format96-well microplate96-well microplate
Intended UseAssay for the qualitativedetection of total anti-HAVantibodies in human serum orplasma.Assay for the qualitativedetection of total anti-HAVantibodies in human serum orplasma.
Required sample volume50 μl50 μl
Specimen StorageRequirementsSamples may be stored at2-8 °C for up to 24 hours.Samples may be stored at2-8 °C for up to 24 hours.
CalibratorReferenced to WHO Anti-Hepatitis A Immunoglobulin2nd International Standard.Referenced to WHO Anti-Hepatitis A Immunoglobulin2nd International Standard.
WavelengthDual wavelength reading at450 nm and 615/630 nm.Dual wavelength reading at450 nm and 630 nm.
Interpretation of resultsObtained absorbance valuereadings for patientspecimens are compared tothe cut-off value determinedby the mean of the calibratorObtained absorbancereadings for patientspecimens are compared to acut-off value determined fromthe mean of the calibrator

Table 3: Similarities between kits with regard to function and use

Table 4: Differences between kits with regard to function and use

Differences in Functionand UseMONOLISAT™ anti-HAV EIACatalog# 72496ETI-AB-HAVK PLUSCatalog# P001926
SpectrophotometricVerification of Sample andReagent PipettingPossible (but optional)NA

{6}------------------------------------------------

Image /page/6/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and distributes products for the life science research and clinical diagnostics markets. The logo is in white text on a black background. The text is bold and sans-serif. The logo is simple and recognizable.

EXPECTED VALUES

Healthy individuals

The expected results of the MONOLISA™ Anti-HAV EIA assay were determined in presumably healthy individuals from the Mid-west US (St Louis, Missouri), the Western US (California and Washington) and from Europe (Parma, Italy).

In the Mid-west, the population was 55% female and 45% male, with ages ranging from 1 to 96 years. 48% (134) were pediatric specimens.

The majority of the subjects were White/Caucasian (64%), and 32% were black or African American; for 4% data were not available.

In this study, 41 % were found reactive for Anti-HAV total antibodies, and 57% were found nonreactive.

In the Western US, 73% were from California, 27% were from Washington. The population was 56% female and 44% male, and their ages ranged from 15 to 90 years.

In this population, 38% were found reactive for Anti-HAV total antibodies, and 62% were found nonreactive.

In Europe, the population was 50% female and 50% male, with ages ranging from 18 to 87 years. In this group, 69% were found reactive for Anti-HAV total antibodies and 31% were found nonreactive.

The expected results for the US and for presumably healthy individuals living in Europe are presented below (Tables 5, 6 and 7).

Table 5: Expected Results for MONOLISA™ Anti-HAV EIA in subjects from the Midwest US (N= 280)

MONOLISA™ Anti-HAV EIA
Age RangeGenderReactiveBorderlineNonreactiveTotal
N%N%N%
< 10Female1028.6%0N/A2571.4%35
Male718.4%25.3%2976.3%38
10-19Female1436.8%25.3%2257.9%38
Male939.1%0N/A1460.9%23
20-29Female360.0%0N/A240.0%5
Male3100.0%0N/A0N/A3
30-39Female550.0%0N/A550.0%10
Male333.3%0N/A666.7%9
40-49Female323.1%0N/A1076.9%13
Male450.0%0N/A450.0%8
50-59Female1055.6%0N/A844.4%18
Male847.1%0N/A952.9%17
60-69Female857.1%0N/A642.9%14
Male430.8%0N/A969.2%13
70-79Female666.7%111.1%222.2%9
Male583.3%0N/A116.7%6
80-89Female969.2%0N/A430.8%13
Male350.0%0N/A350.0%6
>=90Female0N/A0N/A0N/A0
Male150.0%0N/A150.0%2
Total11541.1%51.8%16057.1%280

{7}------------------------------------------------

Image /page/7/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and distributes products for the life science research and clinical diagnostics markets. The logo is white text on a black background. The text is in a bold, sans-serif font.

MONOLISAT™ Anti-HAV EIA
GenderReactiveBorderlineNonreactive
Age RangeN%N%N%Total
<19Female360.0%0N/A240.0%5
Male120.0%0N/A480.0%5
20-29Female1142.3%0N/A1557.7%26
Male520.8%0N/A1979.2%24
30-39Female1050.0%0N/A1050.0%20
Male527.8%0N/A1372.2%18
40-49Female633.3%0N/A1266.7%18
Male1045.5%0N/A1254.5%22
50-59Female1538.5%12.6%2359.0%39
Male523.8%0N/A1676.2%21
60-69Female650.0%0N/A650.0%12
Male433.3%0N/A866.7%12
70-79Female111.1%0N/A888.9%9
Male150.0%0N/A150.0%2
80-89Female6100%0N/A0N/A6
Male375.0%0N/A125.0%4
>=90Female0N/A0N/A1100.0%1
Male0N/A0N/A0N/A0
UnknownFemale0N/A0N/A1100.0%1
Total9237.6%10.4%15262.0%245

Table 6: Expected Results for MONOLISA™ Anti-HAV EIA in subjects from the Western US (N= 245)

{8}------------------------------------------------

Image /page/8/Picture/0 description: The image shows the logo for BIO-RAD. The logo is white text on a black rounded rectangle. The text is in all caps and bolded.

MONOLISAT™ Anti-HAV EIA
Age RangeGenderReactiveBorderlineNonreactiveTotal
N%N%N%
< 19Female0N/A0N/A1100.0%1
Male0N/A0N/A1100.0%1
20-29Female133.3%0N/A266.7%3
Male0N/A0N/A2100.0%2
30-39Female114.3%0N/A685.7%7
Male228.6%0N/A571.4%7
40-49Female733.3%0N/A1466.7%21
Male315.8%0N/A1684.2%19
50-59Female1045.5%0N/A1254.5%22
Male1451.9%0N/A1348.1%27
60-69Female3786.0%0N/A614.0%43
Male2385.2%0N/A414.8%27
70-79Female3196.9%0N/A13.1%32
Male3286.5%0N/A513.5%37
80-89Female13100.0%0N/A0N/A13
Male23100.0%0N/A0N/A23
Total19769.1%0N/A8830.9%285

:

Table 7: Expected Results for MONOLISA™ Anti-HAV EIA in subjects from Italy, Europe (N= 285)

:

{9}------------------------------------------------

Image /page/9/Picture/0 description: The image shows the logo for Bio-Rad. The logo is white text on a black rounded rectangle. The text is bold and sans-serif.

Adult Subjects At High Risk For Viral Hepatitis:

Expected results of asymptomatic prospective high-risk subjects, determined from a multi-center study in the US and in Europe, are reported in the following tables.

A total of 230 US subjects were at high risk for viral hepatitis including intravenous drug users (N= 55), homosexual males (N=15), sex workers (N=39), prison history (N= 92), high-risk sex partners (N=25), and high-risk occupation/health care workers (N=4). Many had more than 1 high-risk behavior or risk factor. Subjects were from Los Angeles, CA, (86.5%), Santa Ana, CA (4.3%), or Miami, FL (9.1%). The group was Caucasian (7.4%), Black or African American (74.3%), Hispanic or Latino (15.2%), Asian (0.4%), Native Hawaiian or other Pacific Islander (0.4%), and American Indian or Alaska native (0.9%), with the remaining (1.3%) represented by multiple ethnic groups.

Of these subjects, 81% were male and 19% were female, and they ranged in age from 18 to 70 years (mean age of 45). The data are reported in Table 8.

The percent of Anti-HAV reactive results with MONOLISA™ Anti-HAV EIA in this high-risk asymptomatic population was 53%.

The European group (N= 62) was 87% male and 13% female, and ranged in age from 21 to 75 years (mean age of 40). It consisted of intravenous drug users (30), subjects who had clotting factor disorders (7) and MSM patients (25). The data are reported in Table 9.

The percent of Anti-HAV reactive results with MONOLISA™ Anti-HAV EIA in this high-risk asymptomatic population was 45%.

MONOLISAT™ Anti-HAV EIA
AgeGenderReactiveBorderlineNonreactive
RangeN%N%N%Total
< 19Female1100.0%0N/A0N/A1
Male1100.0%0N/A0N/A1
20-29Female133.3%0N/A266.7%3
Male150.0%0N/A150.0%2
30-39Female457.1%0N/A342.9%7
Male1233.3%0N/A2466.7%36
40-49Female1562.5%0N/A937.5%24
Male3743.5%11.2%4755.3%85
50-59Female685.7%0N/A114.3%7
Male3160.8%12.0%1937.3%51
60-69Female1100.0%0N/A0N/A1
Male990.0%0N/A110.0%10
70-79Female0N/A0N/A0N/A0
Male2100.0%0N/A0N/A2
Total12152.6%20.9%10746.5%230

Table 8: Expected results for MONOLISA™ Anti-HAV EIA in the US High Risk Group for Viral Hepatitis A (N=230)

{10}------------------------------------------------

Image /page/10/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and distributes products for the life science research and clinical diagnostics markets. The logo is in white text on a black background. The text is bold and sans-serif.

Table 9: Expected results for MONOLISA™ Anti-HAV EIA in the European High Risk Group for Viral Hepatitis A (N=62)

AgeRangeGenderReactiveBorderlineNonreactiveTotal
N%N%N%
< 19Female0N/A0N/A0N/A0
Male0N/A0N/A0N/A0
20-29Female0N/A0N/A5100.0%5
Male0N/A0N/A11100.0%11
30-39Female150.0%0N/A150.0%2
Male535.7%0N/A964.3%14
40-49Female1100.0%0N/A0N/A1
Male964.3%0N/A535.7%14
50-59Female0N/A0N/A0N/A0
Male981.8%0N/A218.2%11
60-69Female0N/A0N/A0N/A0
Male150.0%0N/A150.0%2
70-79Female0N/A0N/A0N/A0
Male2100.0%0N/A0N/A2
>80Female0N/A0N/A0N/A0
Male0N/A0N/A0N/A0
Total2845.2%0N/A3454.8%62

.

.

{11}------------------------------------------------

Image /page/11/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and supplies products for the life science research and clinical diagnostics markets. The logo is in a rounded rectangle shape with the words "BIO-RAD" in bold, white letters against a black background. The logo is simple and recognizable, and it is used on the company's products, website, and marketing materials.

PERFORMANCE CHARACTERISTICS

Clinical performance

A multi-center prospective and retrospective study was conducted to evaluate the clinical performance of the MONOLISA™ Anti-HAV EIA assay among individuals with signs or symptoms and those at high risk of Hepatitis infection. Specimens were collected in 3 different geographical areas: 404 specimens were collected in the US and 928 were collected in Europe (France and Italy).

The US population consisted of 174 subjects with signs and symptoms of Hepatitis.

Of these, 60% were male and 40% were female, and they ranged in age from 17 to 72 years (mean age of 38). The group was Caucasian (13.2%), Black or African American (4.6%), Hispanic or Latino (2.9%), and Asian (41.9%), with 1.1% represented by multiple ethnic groups. The remaining 36.8% were unknown. Among these 174 subjects, 23 (13.2%) were pediatric samples.

The 230 subjects from the high-risk group for Hepatitis A include intravenous drug users (N= 55), homosexual males (N=15), sex workers (N=39), prison history (N= 92), high-risk sex partners (N=25), and high-risk occupation/health care workers (N=4). Many had more than 1 high-risk behavior or risk factor. The group was Caucasian (7.4%), Black or African American (74.3%), Hispanic or Latino (15.2%), Asian (0.4%), Native Hawaiian or other Pacific Islander (0.4%), and American Indian or Alaska native (0.9%), with the remaining (1.3%) represented by multiple ethnic groups. Of these, 81% were male and 19% were female, and they ranged in age from 18 to 70 years (mean age of 45). Among these 230 subjects, 2 (0,9%) were pediatric samples.

The European population consisted of 252 specimens collected from patients with signs and symptoms of Hepatitis. Of these, 51% were male and 49% were female, and they ranged in age from 1 to 105 years (mean age of 53).

Sixty-two (62) specimens were collected from a population at high risk for hepatitis composed of intravenous drug users (30), subjects who had clotting factor disorders (7) and MSM patients (25). The group was 87% male and 13% female, and ranged in age from 21 to 75 years (mean age of 40).

Three hundred and forty five (345) specimens were from an asymptomatic hospitalized population. Of these, 51% were male and 49% were female, and they ranged in age from 18 to 87 years (mean age of 59).

Thirfty four (34) specimens were from healthcare workers (for HAV pre-vaccination screening). One hundred and fifty one (151) patients had recovered HAV infection.

Among these 844 european samples, 35 (4.1%) were from pediatric subjects.

Vaccinated subjects:

Sixty-two (62) pre- and post-vaccination samples from 38 individuals were tested. Fourteen (14) individuals were enrolled in a vaccination program. They received the TWINRIX® vaccine, a combined Hepatitis A and Hepatitis B vaccine from GlaxoSmithKline. A pre-vaccination sample was collected the day of the first vaccination dose. A second sample was collected before the second vaccination dose was injected (one month after the first dose). A third dose of vaccine was scheduled 6 months after the first injection. The sample after the third vaccination dose was not available.

{12}------------------------------------------------

Image /page/12/Picture/0 description: The image is a logo for Bio-Rad. The logo is white text on a black background. The text is in a bold, sans-serif font. The logo is in a rounded rectangle shape.

Twenty (20) samples were collected from 10 subjects, aged 24 to 45 years, who had received the HAVRIX® vaccine. These subjects received HAVRIX® 1440, an inactivated Hepatitis A vaccine from GlaxoSmithKline, in a two-dose schedule (at 0 and 6 to 12 months). For each subject, a preand a post-vaccination specimen was obtained. All post-vaccination samples were obtained 4 weeks after vaccination.

Fourteen (14) purchased post-vaccination samples were from individuals vaccinated with HAVRIX® and 6 were from individuals vaccinated with VAQTA® from Merck &Co.

Percent Agreement

The results obtained with MONOLISA™ Anti-HAV EIA were compared with the results obtained using the comparative assay.

The positive and negative percent agreements and the 95% exact confidence between MONOLISA™ Anti-HAV EIA and the comparative assay were calculated.

To determine the percent agreement on borderline results the following criteria were used:

  • Specimens that were borderline with the comparative assay and reactive with MONOLISA™ Anti-HAV EIA were considered as false positives for MONOLISA™ Anti-HAV EIA assay.

  • Specimens that were borderline with the comparative assay and nonreactive with MONOLISA™ Anti-HAV EIA were considered as false negatives for MONOLISA™ Anti-HAV EIA.

The results obtained with the US specimens and with the European specimens are presented in the following tables.

Table 10: MONOLISA™ Anti-HAV EIA versus the comparative assay Results in the US
Population (N=404)would and the country of the contract and the contrôleant with and
Subject categoryComparative assay:PositiveMONOLISA™ Anti-HAV EIAComparative assay:BorderlineMONOLISA™ Anti-HAVEIAComparative assay:NegativeMONOLISA™ Anti-HAV EIATotal
RBRDNRRBRDNRRBRDNR
Subjects with signsand symptoms123021001344174
Subjects with highrisk for Hepatitis1140021141107230
Total237023b1d1c54a151404
Positive percentagreement95% ExactConfidence intervalNegative percentagreement95% ExactConfidence interval
Total98,8%(237/240)96.4 - 99.792.6%(151/163)87.5 - 96.1

R : Reactive, NR : Nonreactive, BRD : Borderline

8 : the Borderline results with MONOLISA™ Anti-HAV EIA were considered as false positives.

b: the specimens that were Borderline with the comparative assay and reactive with MONOLISA'™ Anti-HAV EIA were considered as false positives with MONOLISA™ Anti-HAV EIA.

4: the results that were borderline with both the MONOLISA™ Anti-HAV EIA and with the comparative assay were not included in the negative agreement or the positive agreement calculations.

ci the specimens that were Borderline with the comparative with MONOLISA™ Anti-HAV EIA were considered as false negative with MONOLISA™ Anti-HAV EIA.

{13}------------------------------------------------

Image /page/13/Picture/0 description: The image shows the logo for BIO-RAD. The logo is white text on a black background. The text is in a bold, sans-serif font. The logo is in a rounded rectangle shape.

Table 11: Comparison of Results for MONOLISA™ Anti-HAV EIA versus the
comparative assay in the European Population (N= 844)
SubjectcategoryComparative assay:PositiveMONOLISA™ Anti-HAV EIAComparative assay:BorderlineMONOLISA™ Anti-HAV EIAComparative assay:NegativeMONOLISA™ Anti-HAV EIATotal
RBRDNRRBRDNRRBRDNR
Generalhospitalizedpopulation2360100000108345
Sign /Symptoms ofHepatitis190000211157252
Subjectswith highrisk forHepatitis2800000003462
Healthcareworkers600000002834
Infected/recoveredHAV15000100000151
Total610011b2d1c11a227844
Positive percentagreement95% ExactConfidence intervalNegative percentagreement95% ExactConfidence interval
Total99.7%(610/612)98.8 – 99.998.7%(227/230)96.2 – 99.7

R : Reactive, NR : Nonreactive, BRD : Borderline

a : the Borderline result with MONOLISA™ Anti-HAV EIA was considered as false positive

b the specimen that was Borderline with the comparative assay and reactive with MONOLISA™ Anti-HAV EIA was considered as false positive with MONOLISA™ Anti-HAV EIA.

^ the specimen that was Borderline with the comparative assay and nonreactive with MONOLISA™ Anti-HAV EIA was considered as false negative with MONOLISA™ Anti-HAV EIA.

d :the 2 borderline results with both MONOLISA™ Anti-HAV EIA and with the comparative assay were not included in the calculation of the negative agreement or the positive agreement.

Acute HAV Infection:

Among the retrospective samples, 84 were from subjects with a medical history and laboratory results indicative of acute Hepatitis A. The subjects included 56% male, 37% female; the gender was not available for 7%. The mean age was 21, and subjects ranged from 1 to 55 years. Among them 39 were pediatric subjects.

The results are presented in the following table:

{14}------------------------------------------------

Image /page/14/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and distributes products for the life science research and clinical diagnostics markets. The logo is in black and white, with the words "BIO-RAD" in bold, sans-serif font. The logo is enclosed in a rounded rectangle. The logo is simple and recognizable.

Table 12: Comparison of Results for MONOLISA™ Anti-HAV EIA versus the comparative assay on Acute HAV infection in the adult and pediatric European Population (N= 84) :

Comparative assay:PositiveMONOLISA™ Anti-HAV EIAComparative assay:BorderlineMONOLISA™ Anti-HAV EIAComparative assay:NegativeMONOLISA™ Anti-HAV EIATotal
RBRDNRRBRDNRRBRDNR
Adults450000000045
Pediatrics390000000039
Total840000000084

R: Reactive, NR: Nonreactive, BRD : Borderline

The positive agreement was 100% (84/84) with a 95% exact confidence interval of 96.5% to 100%.

Performance of MONOLISA™ Anti-HAV EIA in pediatric subjects:

Sixty (60) pediatric samples were tested during the US and European clinical studies in addition to the 39 pediatric samples from acute HAV infection.

Among the US population, 23 had signs and symptoms of hepatitis and 2 were from the high risk group. In the European population, 3 belonged to the general hospitalized population, 22 had signs and symptoms of hepatitis, 2 were from the high risk group, 3 were healthcare workers, 5 had recovered from Hepatitis A infection. The results from these pediatric samples are summarized in the following table.

{15}------------------------------------------------

Image /page/15/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and distributes products for the life science research and clinical diagnostics markets. The logo is a black rectangle with rounded corners, and the words "BIO-RAD" are written in white, bold letters inside the rectangle. The letters are large and easy to read, and the overall design is simple and modern.

Table 13 : Comparison of Results for MONOLISA™ Anti-HAV EIA versus the
comparative assay in the Pediatric European and US Population (N= 60)
SubjectcategoryComparative assay:PositiveComparative assay:BorderlineComparative assay:NegativeTotal
RBRDNRRBRDNRRBRDNR
EuropeanPediatrics1600000001935
USPediatrics1300000011125
Total2900000013060
Positive percentagreement95% ExactConfidence intervalNegative percentagreement95% ExactConfidence interval
Total100%(29/29)90.2 - 10096.8%(30/31)83.3 - 99.9

R: Reactive, NR: Nonreactive, BRD: Borderline

Including the combined US and European Sites, the positive percent agreement of the MONOLISA™ Anti-HAV EIA with the comparative anti-HAV assay was 99.5% (931/936) with a 95% exact confidence interval of 98.8% to 99.8%. The negative percent agreement of the MONOLISA™ Anti-HAV EIA with the comparative anti-HAV assay was 96.2% (378/393) with a 95% exact confidence interval of 93.8% to 97.9%.

Study On Vaccinated Subjects:

The HAV antibody response to vaccination was evaluated with 3 different vaccines that are currently licensed in the US: VAQTA® from Merck & Co, HAVRIX® 1440 from Glaxo SmithKline and TWINRIX® from Glaxo SmithKline.

For VAQTA® vaccine, 6 post-vaccination samples from US subjects were available.

For HAVRIX® vaccine, 10 matched sets of pre- and post-vaccination samples from European subjects and 8 post-vaccination samples from US subjects were available.

For TWINRIX® vaccine, 14 matched sets of pre-vaccination and post first dose samples from European individuals were available.

The following results were obtained:

{16}------------------------------------------------

Image /page/16/Picture/0 description: The image shows the logo for Bio-Rad. The logo is white text on a black rounded rectangle. The text is in a bold, sans-serif font.

Comparative assay:PositiveMONOLISA™ Anti-HAV EIAComparative assay:BorderlineMONOLISA™ Anti- HAV EIAComparative assay:NegativeMONOLISA™ Anti-HAV EIATotal
RBRDNRRBRDNRRBRDNR
VAQTAPost-vaccination6000000006
HAVRIXPre-vaccination000000001010
HAVRIXPost-vaccination180000000018
TWINRIXPre-vaccination1000001*01214
TWINRIXPost 1st injection91000200214

Table 14: MONOLISA™ Anti-HAV EIA Results on Vaccinated Subjects versus the comparative assay - All testing sites

R : reactive, NR : Nonreactive , BRD : Borderline

  • Result close to the cutoff value (CO/S=1.2)

In pre-vaccination samples, MONOLISA™ Anti-HAV EIA was in overall agreement with the comparative assay for 21/22 (95.5%) of samples tested.

For TWINRIX® vaccine on post first dose vaccination, MONOLISA™ Anti-HAV EIA demonstrated reactivity in 9/14 (64.3%) samples. The reference method demonstrated reactivity in 10/14 (71.4%) samples.

For HAVRIX® post-vaccination samples, MONOLISA™ Anti-HAV EIA demonstrated reactivity in 18/18 (100%) samples. The reference method demonstrated reactivity in 18/18 (100%) samples.

For VAQTA® post-vaccination samples, MONOLISA™ Anti-HAV EIA demonstrated reactivity in 6/6 (100%) samples. The reference method demonstrated reactivity in 6/6 (100%) samples.

{17}------------------------------------------------

Image /page/17/Picture/0 description: The image shows the logo for Bio-Rad. The text "BIO-RAD" is in white, bold, sans-serif font. The text is set against a black, rounded rectangle.

Seroconversion Panels

Six commercially available HAV seroconversion panels were tested using MONOLISA™ Anti-HAV EIA and the FDA approved comparative assay to determine the sensitivity of the assay. The results are summarized in the following table:

Panel IDMONOLISA™ Anti-HAV EIAComparative assay
Post bleed day of firstreactive resultPost bleed day of firstreactive resultDifference in Days toReactive result
07467A000
60160K000
HAV01000
RP-00406- 6
RP-013880
PHT90216160

Table 15: MONOLISA™Anti-HAV EIA Seroconversion Panels Results :

The sensitivity of the MONOLISA™ Anti-HAV EIA was equivalent to or more sensitive than the comparative assay in the six seroconversion panels tested.

Cross Reactivity Study

The potential for cross reactivity to other disease states, or viruses was evaluated for the MONOLISA™ Anti-HAV EIA Assay and the comparative assay.

In addition, samples containing rheumatoid factors, auto-antibodies, anti-mouse antibodies were tested.

In total, 255 specimens (including both serum and plasma) from 16 groups of potential crossreactivity were tested. FDA approved methods were used to confirm the disease state of each specimen.

The results are summarized in the following table.

{18}------------------------------------------------

Image /page/18/Picture/0 description: The image shows the logo for Bio-Rad. The logo is white text on a black rounded rectangle. The text is in a bold, sans-serif font.

Table 16: Potential cross reactivity study

Clinical ConditionComparative assay PositiveMONOLISA™ Anti-HAV EIAComparative assay BRDMONOLISA™ Anti-HAV EIAComparative assay NegativeMONOLISA™ Anti-HAV EIATotal
RBRDNRRBRDNRRBRDNR
Hepatitis C (HCV)70100000715
Hepatitis B (HBV) HBs Ag90000000615
Hepatitis B (HBV) anti HBc100000031115
Human Immunodeficiency Virus (HIV)60110000715
Epstein Barr Virus (EBV) IgG100100001315
Epstein Barr Virus (EBV) IgM150000000015
Cytomegalovirus (CMV) IgG60000000915
Cytomegalovirus (CMV) IgM70000000815
Rubella IgG50100000915
Toxoplasmosis IgG100000000515
Toxoplasmosis IgM82000100415
Mumps IgG300000101115
Varicella Zoster Virus(VZV) IgG100000001415
Varicella Zoster Virus(VZV) IgM60010100715
Anti Nuclear Antibody (ANA)70000100715
Human Anti Mouse Antibody (HAMA)200001001215
Rheumatoid Arthritis120000000315
Total1152330441123255

7 samples were discrepant: 4 reactive on MONOLISA™ Anti-HAV EIA, nonreactive on comparative assay and 3 were
nonreactive on MONOLISA™ Anti-HAV EIA and reactive on comparative

{19}------------------------------------------------

Image /page/19/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and distributes products for the life science research and clinical diagnostics markets. The logo is a black rounded rectangle with the words "BIO-RAD" in white, bold, sans-serif font. The letters are tightly spaced and the "+" symbol is used in place of the hyphen.

Precision Study

Within-Laboratory Precision Study :

A 21-member panel was tested: serum samples with the 6 corresponding plasma samples (EDTA K2, EDTA K3, Sodium Citrate, Sodium Heparin, Lithium heparin, ACD) at 3 different levels (1 negative, 1 negative near the cutoff, 1 low positive near the cutoff) were tested on 1 lot, in duplicate, in 2 different runs per day (am and pm), by the same operator for a period of 20 days. The data were analyzed following the CLSI guidance EP5A2. The mean ratio, the Standard Deviation (SD) and percent coefficient of variation (%CV) were calculated for each panel member.

The data summary is shown in the following tables.

Panel MemberNMeanCO/SWithin run¹Between Run ²Between Day ³Total ⁴
SDCV (%)SDCV (%)SDCV (%)SDCV (%)
Negative Control C0400.282NANA0.026.4%0.015.0%0.028.1%
Positive Control C1404.093NANA0.5413.5%0.000.0%0.4912.3%
Serum 1800.3950.024.1%0.036.3%0.011.9%0.037.7%
EDTA K2 1800.3790.025.0%0.036.8%0.011.4%0.038.6%
EDTA K3 1800.3760.013.4%0.049.3%0.000.0%0.049.9%
Sodium Citrate 1800.3870.0410.1%0.013.3%0.025.1%0.0511.8%
Sodium Heparin 1800.3630.013.4%0.037.6%0.000.0%0.038.3%
Lithium Heparin 1800.3640.013.4%0.037.0%0.013.5%0.038.5%
ACD 1800.4020.024.3%0.0410.4%0.000.0%0.0511.3%
Serum 2800.6910.034.9%0.069.6%0.012.3%0.0611.0%
EDTA K2 2800.6570.023.2%0.056.6%0.011.8%0.057.5%
EDTA K3 2800.6860.034.9%0.068.2%0.000.0%0.079.5%
Sodium Citrate 2800.6360.033.7%0.057.0%0.034.8%0.069.2%
Sodium Heparin 2800.6280.023.1%0.046.0%0.034.2%0.067.9%
Lithium Heparin 2800.6850.056.6%0.068.6%0.000.0%0.0810.8%
ACD 2800.7460.045.6%0.056.4%0.034.7%0.079.7%
Serum 3801.5060.064.2%0.149.4%0.000.4%0.1510.3%
EDTA K2 3801.2610.074.7%0.117.0%0.074.8%0.159.7%
EDTA K3 3801.2570.042.4%0.096.0%0.053.6%0.117.4%
Sodium Citrate 3801.4620.085.1%0.138.7%0.074.9%0.1711.2%
Sodium Heparin 3801.3800.117.5%0.128.0%0.053.4%0.1711.4%
Lithium Heparin 3801.3460.085.6%0.117.1%0.031.8%0.149.2%
ACD 3801.3440.053.4%0.085.6%0.096.0%0.138.9%

Table 17: MONOLISA™ Anti-HAV EIA Precision Results by Panel Member Cutoff to Signal (CO/S)

NA : Not Applicable

1 Within Run: variability of the assay performance from replicate to replicate

2Between Run: variability of the assay performance from Run to Run

3 Between Day: variability of the assay performance from Day to Day

*Total :total variability of the assay performance includes within run, between run and between day.

{20}------------------------------------------------

Image /page/20/Picture/0 description: The image shows the logo for Bio-Rad. The logo is in white text on a black background. The text is bold and sans-serif.

Reproducibility Study:

A 6 member panel consisting of diluted plasma specimens (negative and different levels of positive) was tested in triplicate, once a day for 3 days on 3 lots* of MONOLISA™ Anti-HAV EIA at 3 separate clinical trial sites. Each panel was coded with a different number on each day tested in order to blind the operator to the expected value of the sample.

*:3 different lots were used at the Bio-Rad site and 2 lots were used on each of the external sites.

The data from all reagent lots and sites were combined to obtain Standard Deviation (SD) and percent coefficient of variation (CV) for within run, between lot, between lot, between site and total variance. The data were analyzed according to the principles described in the Clinical Laboratory Standards Institute guidance EP5-A2, revised November 2004 and ISO/TR 22971:2005. The PROC GLM procedure in SAS® was used to estimate the variance components of the model. The model was y = site + lot (site) + day (lot site) + error.

The summaries are shown in the following tables.

Table 18: MONOLISA™ Anti-HAV EIA Reproducibility Results by Panel Member Cutoff to Signal (CO/S)

TestsitePanelMemberNMeanCO/SWithin Run¹Between Day²Between Lot³Total⁴
SD%CVSD%CVSD%CVSD%CV
Site#1P1180.330.013.030.0928.10⁵00.0928.3
P2180.680.034.90.0710.00⁵00.0711.1
P3181.020.055.00.055.20⁵4.10.088.3
P4181.980.094.70.3216.00⁵00.3316.7
P5182.480.187.40.3614.80⁵00.4116.5
P6183.660.236.20.205.50⁵00.38.3
Site#2P1180.350.011.60.024.70.011.20.025.1
P2180.920.033.00.078.00⁵00.088.5
P3181.280.043.50.000.00.010.620.053.6
P4182.320.083.60.208.50.020.950.219.3
P5183.100.134.10.206.40⁵00.237.5
P6184.160.133.20.368.70⁵00.399.3
Site#3P1270.360.014.00.025.40.026.60.039.4
P2270.810.033.40.033.90.067.20.078.8
P3271.270.086.60.043.60.1411.20.1713.5
P4272.160.115.00.052.20.3415.70.3616.6
P5273.090.113.70.154.90.4915.70.5216.9
P6274.470.112.50.337.31.0122.51.0623.8

1 Within Run: variability of the assay performance from replicate to replicate

2Between Day: variability of the assay performance from Day to Day

3 Between Lot: variability of the assay performance from Lot to Lot

"Total : total variability of the assay performance includes within run, between day and between lot.

ً Negative variances were rounded to zero, per statistical convention.

{21}------------------------------------------------

Image /page/21/Picture/0 description: The image shows the logo for Bio-Rad Laboratories, a company that manufactures and distributes products for the life science research and clinical diagnostics markets. The logo is in black and white and features the company name in bold, sans-serif font. The letters are white, and the background is black. The logo is enclosed in a rounded rectangle.

Table 19: MONOLISA™ Anti-HAV Reproducibility Summary by Panel Member Cutoff to Signal (CO/S)

Panel MemberNMeanWithin Run1Between Day2Between Lot3Between Site5Total4
SD%CVSD%CVSD%CVSD%CVSD%CV
P1630.350.013.20.0514.80500.012.90.0515.5
P2630.800.033.70.067.120.033.70.1113.40.1316.0
P3631.200.075.50.043.30.108.60.1210.30.1814.9
P4632.150.104.50.209.50.2210.00.000.00.3114.5
P5632.920.144.80.248.30.3211.00.258.70.5017.0
P6634.150.163.80.317.40.7016.90500.7818.9

1 Within Run: variability of the assay performance from replicate to replicate

2Between Day: variability of the assay performance from Day to Day

3Between Lot: variability of the assay performance from Lot to Lot

*Between site: variability of the assay performance from Site to Site

"Total : total variability of the assay performance includes within run, between lot and between site.

Trotal . total variablity of the assuy performance includes within full,
5 Negative variances were rounded to zero, per statistical convention.

Reproducibility study on Negative and Positive Controls:

The negative and positive controls were tested in triplicate, once a day by 3 different operators for 3 days. The data were analyzed according to the principles described in the Clinical Laboratory Standards Institute guidance EP5-A2, revised November 2004 and ISO/TR 22971:2005.

Table 20: MONOLISA™ Anti-HAV EIA Control Reproducibility summary by Operator (CO/S)

SamplesNMeanWithin Run1Between Day2BetweenOperator 3Total4
SD%CVSD%CVSD%CVSD%CV
Negative Control270.300.012.60.028.30500.038.7
Positive Control274.740.163.50.4810.10.071.50.5110.8

1 Within Run: variability of the assay performance from replicate to replicate

2Between Day: variability of the assay performance from Day to Day

3 Between operator: variability of the assay performance from Operator to Operator

*Total :total variability of the assay performance includes within run, between day and between Operator.

5 Negative variances were rounded to zero, per statistical convention.

{22}------------------------------------------------

Image /page/22/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features the department's name in a circular arrangement around a stylized eagle emblem. The eagle is depicted with three wavy lines forming its body and wings, symbolizing the department's mission to protect and promote the health and well-being of Americans.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Manuela Kaul RA-Manager Bio-Rad France 3, Boulevard Raymond Poincaré 92430 Marnes-la-Coquette, France

MAY - 3 2007

Re: K063318

Trade/Device Name: MONOLISA™ Anti-HAV EIA Regulation Number: 21 CFR 866.3310 Regulation Name: Hepatitis A Virus (HAV) Serological Reagents Regulatory Class: Class II Product Code: LOL Dated: March 21, 2007 Received: April 3, 2007

Dear Ms. Kaul:

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. The general controls provisions of the Act include requirements for annual registration. Ilsting of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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 requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); and good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820).

{23}------------------------------------------------

This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 594-3084. Also, please note the regulation entitled. "Misbranding by reference to premarket notification" (21CFR Part 807.97), Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 443-6597 or at its Internet address http://www.fda.gov/cdrh/dsma/dsmamain.html.

Sincerely yours,

Kaily att

Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostic Devices Evaluation and Safety Center for Devices and Radiological Health

Enclosure

{24}------------------------------------------------

INDICATIONS FOR USE STATEMENT

510(k) Number : K 063318

MONOLISA™ Anti-HAV EIA Device Name:

Indications for Use:

The MONOLISA™ Anti-HAV EIA is an in vitro enzyme immunoassay kit intended for use in the qualitative detection of total antibodies (IgG and IgM) to Hepatitis A Virus (anti-HAV) in human (adult and pediatric) serum or plasma (EDTA, Heparin, Citrate, ACD). This kit can be used as an aid in the diagnosis of acute or past Hepatitis A Virus (HAV) infection or as an aid in the identification of HAVsusceptible individuals for vaccination. However, any diagnosis should take into consideration the patient's clinical history and symptoms, as well as serological data.

Assay performance characteristics have not been established for immunocompromised or immunosuppressed patients, and cord blood or neonatal specimens.

WARNING : This assay is not intended for screening blood or solid or soft tissue donors.

Prescription Use: ___ X (Per 21 CFR 801.109) AND/OR

Over-The-Counter Use: (Optional Format 1-2-96)

(PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of Device Evaluation (ODE)

Faisano

Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety

51000 K063318

§ 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.