The BioPlex® 2200 Rubella and CMV IgM kit is a multiplex flow immunoassay intended for the qualitative detection of IgM antibodies to Rubella and Cytomegalovirus (CMV) in human serum and potassium EDTA or sodium heparin plasma.

The BioPlex 2200 Rubella and CMV IgM kit is intended for use with the Bio-Rad BioPlex 2200 System.

This kit is intended as an aid in the diagnosis of a current or recent Rubella and/or CMV infection, in individuals suspected of having one of the respective disease states including women of child bearing age.

This assay is not FDA cleared or approved for use in testing (screening) blood or plasma donors.

Performance characteristics for the Rubella and CMV IgM assays have not been evaluated in immunosupressed or organ transplant individuals. Performance characteristics of this kit have not been established for use in neonatal screening or for use at point of care facilities.

The BioPlex® 2200 Rubella and CMV IgM Calibrator Set is intended for calibration of the BioPlex 2200 Rubella and CMV IgM Reagent Pack.

The BioPlex 2200 Rubella and CMV IgM Control Set is intended for use as an assayed quality control to monitor the overall performance of the BioPlex 2200 Instrument and BioPlex Rubella and CMV IgM Reagent Pack in the clinical laboratory. The performance of the BioPlex 2200 Rubella and CMV IgM Control Set has not been established with any other Rubella or Cytomegalovirus (CMV) IgM antibody assays.

Device Description

The BioPlex® 2200 Rubella and CMV IgM kit uses multiplex flow immunoassay, a methodology that greatly resembles traditional EIA, but permits simultaneous detection and identification of many antibodies in a single tube. Rubella and CMV IgM test is to detect antibodies to Rubella and Cytomegalovirus (CMV).

Two (2) different populations of dyed beads are coated with cell lysates bearing Rubella or CMV antigens. The BioPlex 2200 System combines an aliquot of patient sample, sample diluent, and bead reagent into a reaction vessel; the mixture is incubated at 37°C. After a wash cycle, anti-human IgM antibody, conjugated to phycoerythrin (PE), is added to the dyed beads, and this mixture is incubated at 37°C. The excess conjugate is removed in another wash cycle, and the beads are re-suspended in wash buffer. The bead mixture then passes through the detector. The identity of the dyed beads is determined by the fluorescence of the dyes, and the amount of antibody captured by the antigen is determined by the fluorescence of the attached PE. Raw data are reported as relative fluorescence intensity (RFI).

Three additional dyed beads, an Internal Standard Bead (ISB), a Serum Verification Bead (SVB) and a Reagent Blank Bead (RBB) are present in each reaction mixture to verify detector response, the addition of serum or plasma to the reaction vessel and the absence of significant non-specific binding in serum or plasma.

The instrument is calibrated using a set of three (3) distinct serum based calibrators. A negative and CMV IgM calibrator is used to calibrate CMV assay, and a negrative and rubella IgM calibrator is used to calibrate the rubella IgM assay, The cul-off value and assignment of the calibrators are determined by performing concordance and Receiver Operator Characteristic (ROC) analysis using the Centaur Rubella IgM and VIDAS CMV IgM predicate results as the standard. For Rubella and CMV, results of ≤ 0.8 Al are negative, 0.9 and 1.0 Al are equivocal and results of ≥ 1.1 Al are reported as positive.

The BioPlex 2200 Rubella and CMV IgM Control Set includes a negative control as well as a CMV IgM positive control and a Rubella IgM positive control. The BioPlex Rubella and CMV IgM Positive Controls are manufactured to give positive results, with values above the cut-off for each specific analyte. The BioPlex Rubella and CMV IgM Negative Control are manufactured to give negative results, with values below the cut-off for each specific analyte. The recommended frequency for performing quality control is once every 24-hour testing period. Performing quality control is also necessary after each new assay calibration and certain service procedures.

AI/ML Overview

Here's an analysis of the provided text regarding the BioPlex® 2200 Rubella and CMV IgM kit, focusing on acceptance criteria and the supporting studies:

Summary of Acceptance Criteria and Device Performance (Based on Method Comparison Studies)

The acceptance criteria for the BioPlex® 2200 Rubella and CMV IgM kit are not explicitly stated as numerical targets in the provided document, but rather implied through comparison to predicate devices and general performance metrics like Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA).

1. Table of Acceptance Criteria and Reported Device Performance

Performance Metric	Acceptance Criteria (Implied)	Reported Device Performance (BioPlex 2200) - Rubella IgM	Reported Device Performance (BioPlex 2200) - CMV IgM
Prospective Study
Positive Percent Agreement (PPA)	Comparable to predicate device	66.7% (2/3) (95% CI: 20.8 - 93.9%)	53.8% (7/13)
Negative Percent Agreement (NPA)	Comparable to predicate device	95.6% (282/295) (95% CI: 92.6 - 97.4%)	97.7% (377/386)
NPA (Pregnant Women - Rubella IgM)	Comparable to predicate device	96.8% (70/71) (95% CI: 92.8 - 99.8%)	N/A (Rubella-specific)
Retrospective Study
PPA (Presumptive Positive)	Comparable to predicate device	96.3% (103/107) (95% CI: 90.3 – 98.5%)	91.3% (209/229) (95% CI: 86.9 - 94.3%)
Matrix Comparison
Slope (vs. Serum)	1.0 ± 0.2	Rubella IgM (EDTA): 1.0369 Rubella IgM (Heparin): 1.0219 CMV IgM (EDTA): Not explicitly stated, but graphically looks good CMV IgM (Heparin): 1.0000	Rubella IgM (EDTA): 1.0369 Rubella IgM (Heparin): 1.0219 CMV IgM (EDTA): Not explicitly stated, but graphically looks good CMV IgM (Heparin): 1.0000
Correlation Coefficient (r) (vs. Serum)	≥ 0.98	Rubella IgM (EDTA): 0.9971 Rubella IgM (Heparin): 0.9976 CMV IgM (EDTA): Not explicitly stated, but graphically looks good CMV IgM (Heparin): 0.9945	Rubella IgM (EDTA): 0.9971 Rubella IgM (Heparin): 0.9976 CMV IgM (EDTA): Not explicitly stated, but graphically looks good CMV IgM (Heparin): 0.9945

Notes on Acceptance Criteria:

The document implies that "comparable performance" or "performed according to its specifications" is the acceptance criterion for many aspects. For quantitative measurements like reproducibility and matrix comparison, specific numerical targets (e.g., %CV ranges, slope 1.0 +/- 0.2, r >= 0.98) are explicitly mentioned and met.
The PPA values for the prospective study, especially for Rubella IgM (66.7%) and CMV IgM (53.8%), seem low initially. However, these are based on a very small number of positive samples (3 for Rubella IgM and 13 for CMV IgM), which leads to wide confidence intervals. The larger retrospective study with presumptive positive samples shows much higher PPA values (96.3% for Rubella IgM and 91.3% for CMV IgM), suggesting strong overall positive agreement when sufficient positive samples are present. The low positive counts in the prospective study are likely due to the low prevalence of acute infections in the "test ordered" population.

2. Sample Sizes Used for the Test Set and Data Provenance

Reproducibility (Internal): 3 panels (serum, EDTA plasma, heparinized plasma). Assayed 2 times in 2 separate daily runs over 20 days (n=80).
Reproducibility (External): 3 panels (serum, EDTA plasma, heparinized plasma). Tested in quadruplicate over 5 days at 3 sites (n=60 replicates per panel member).
Interfering Substances: Specific substances tested at varying concentrations. Number of samples not explicitly stated per substance, but implied to be sufficient to observe interference if present.
Cross-Reactivity: Varied numbers of samples (typically 9-10) for each potential cross-reactant. Samples were known positive for the given cross-reactant and negative by predicate devices.
IgM Detection (DTT treatment): 10 Rubella IgM-positive samples and 10 CMV IgM-positive samples.
Seroconversion Testing: 3 commercial Rubella IgM seroconversion panels (RP001, RP011, RP014) and 1 commercial CMV IgM seroconversion panel (RP003). Each panel consists of multiple bleeds over time.
Expected Values:
- Rubella IgM: 300 samples (US origin).
- CMV IgM: 400 samples (300 US, 100 Europe).
Method Comparison (Prospective Study):
- Rubella IgM: 300 samples (US origin), including 71 pregnant women.
- CMV IgM: 400 samples (300 US, 100 Europe).
Method Comparison (Retrospective Study - Presumptive Positive):
- Rubella IgM: 107 samples.
- CMV IgM: 229 samples.
Matrix Comparison: 20 individual donors for matched serum, potassium EDTA plasma, and sodium heparin plasma samples. Evaluated in replicates of 10.

Data Provenance:

US and Europe: Explicitly mentioned for some Expected Values and Method Comparison studies for CMV IgM. Rubella IgM samples are predominantly US.
Retrospective/Prospective:
- Prospective: Method Comparison study for general population and pregnant women.
- Retrospective: Method Comparison study for presumptive positive samples. Seroconversion panels, interfering substances, and cross-reactivity studies are inherently retrospective in their selection (i.e., using pre-characterized samples). Reproducibility studies used panels.

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

The document does not specify the direct involvement of human experts (e.g., radiologists, pathologists) in establishing the ground truth for this device. Instead, the ground truth for most comparative studies is established by:

Predicate Devices: The ADVIA Centaur® Rubella IgM (K010668) and bioMeriéux VIDAS® CMV IgM (K933549) are used as the reference standard ("predicate results as the standard") for determining cut-off values and for method comparison studies.
FDA Cleared Devices: For the cross-reactivity study, samples were pre-tested by "FDA cleared devices." For adjudication, "two out of three FDA cleared devices" were used.
Commercial Seroconversion Panels: Bio-Rad Laboratories Liquichek™ Rubella IgM and CMV IgM seroconversion panels were used, which are typically well-characterized.

Therefore, the ground truth is based on established, FDA-cleared commercial assays and recognized diagnostic panels, rather than direct expert consensus on individual cases.

4. Adjudication Method for the Test Set

Adjudication by Multiple FDA Cleared Devices: For samples that showed equivocal results by the predicate device in the Method Comparison (Prospective Study), an adjudication method was used: "One sample that was equivocal by the predicate device was adjudicated by two out of three FDA cleared devices." and "Two samples that were equivocal by the predicated by two out of three HDA cleared devices." This is a form of 2-out-of-3 or 3-out-of-3 adjudication against external reference methods.
No explicit 2+1, 3+1, or similar human expert adjudication is mentioned in the context of interpretation of images or clinical cases. The adjudication is against other laboratory tests.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

No MRMC comparative effectiveness study was mentioned or performed. This device is an in vitro diagnostic (IVD) immunoassay kit, not an AI-assisted diagnostic imaging device or tool that involves human "readers" in the traditional sense of interpreting complex data like medical images. Its performance is evaluated against other laboratory assays.

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

The BioPlex® 2200 system operates as a standalone automated immunoassay. The performance studies described (reproducibility, interfering substances, cross-reactivity, seroconversion, method comparison) inherently evaluate its "algorithm only" or automated performance without human intervention in the interpretation process of individual test results. The device provides quantitative results (RFI) which are then interpreted qualitative (negative, equivocal, positive) based on predefined cut-offs.

7. The Type of Ground Truth Used

The ground truth primarily used is "predicate devices" or "FDA cleared devices."

For method comparison studies, the results from the ADVIA Centaur® Rubella IgM and bioMeriéux VIDAS® CMV IgM predicate devices served as the gold standard.
For cross-reactivity, samples confirmed positive for specific conditions by other FDA cleared devices were used.
For seroconversion, commercial seroconversion panels were used, which have well-characterized profiles.
The calibration itself uses concordance and Receiver Operator Characteristic (ROC) analysis with predicate device results.

8. The Sample Size for the Training Set

The document does not explicitly describe a separate "training set" in the context of machine learning or AI algorithm development. This is an immunoassay kit, where the "training" involves the development and calibration of the assay.

The calibration process is described using "a set of three (3) distinct serum based calibrators." The cut-off values are determined by "performing concordance and Receiver Operator Characteristic (ROC) analysis using the Centaur Rubella IgM and VIDAS CMV IgM predicate results as the standard." While not a "training set" in the AI sense, this calibrator set and the analysis with predicate results serve a similar function in establishing the assay's operational parameters.

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

As noted above, there isn't a "training set" in the typical AI sense. For the establishment of assay calibration and cut-offs:

Predicate Device Results: The ground truth for defining the cut-off values and calibrators was established by comparing the BioPlex 2200's performance against the established results of the predicate devices (ADVIA Centaur® Rubella IgM and bioMeriéux VIDAS® CMV IgM) using concordance and ROC analysis. This means the clinical and analytical performance of those predicate devices, which are already FDA cleared, serve as the reference for tuning the BioPlex 2200's output.

Summary

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K092587

510(k) Summary for Bio-Rad Laboratories, Inc. BioPlex® 2200 Rubella and CMV IgM

DEC - 3 2010

1. APPLICANT/SPONSOR

Bio-Rad Laboratories, Inc. BioPlex Division 5500 East Second Street Benicia, CA 94510

Contact Person:	Juang Wang
Telephone:	510-741-4609

Date Prepared: November 30, 2010

2. DEVICE NAME

Proprietary Name:	BioPlex® 2200 Rubella and CMV IgM KitBioPlex® 2200 Rubella and CMV IgM Calibrator SetBioPlex® 2200 Rubella and CMV IgM Control Set
Common/Usual Name:	Multi-Analyte Detection System for Rubella IgM andCytomegalovirus (CMV) IgM
Classification Name:	Rubella virus serological reagentsCytomegalovirus serological reagentsCalibrator, multi-analyte mixtureSingle analyte controls, all kinds (assayed and unassayed)

3. Predicate Devices

ADVIA Centaur® Rubella IgM . K010668 bioMeriéux VIDAS® CMV IgM
. K933549

4. DEVICE DESCRIPTION

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BioPlex® 2200 Rubella and CMV IgM Kit Components

The BioPlex 2200 Rubella and CMV IgM kit (665-1751) contains supplies sufficient for 100 tests.

Vial	Description
Bead Set	One (1) 10 mL vial containing two (2) different populations of dyed beads coated with lysates of Rubella and CMV; an Internal Standard Bead (ISB), a Serum Verification Bead (SVB) and a Reagent Blank Bead (RBB) in buffer with Glycerol and protein stabilizers (bovine). ProClin ® 300 (0.3%) and sodium azide (<0.1%) as preservatives.
Conjugate	One (1) 5 mL vial, containing phycoerythrin-conjugated donkey polyclonal anti-human IgM antibody and phycoerythrin -conjugated murine monoclonal anti-human FXIII antibody, in buffer with stabilizers (bovine and equine). ProClin ® 300 (0.3%), sodium benzoate (0.1%) and sodium azide (<0.1%) as preservatives.
Sample Diluent	One (1) 10 mL vial, containing goat anti-human IgG antibody and protein stabilizers (bovine and equine) in buffer. ProClin ® 300 (0.3%) and sodium azide (<0.1%) as preservatives.

Additional Required Items. Available from Bio-Rad:
----------------------------------------------------	--	--	--

Catalog #	Description
663-1701	BioPlex 2200 Rubella and CMV IgM Calibrator Set: Three (3) 0.5mL vials containing human IgM antibodies to Rubella and CMV, ina human serum matrix made from defibrinated plasma. Allantibodies are derived from human disease state plasma. Allcalibrators contain ProClin® 300 (0.3%), sodium benzoate (0.1%)and sodium azide (<0.1%) as preservatives.
663-1731	BioPlex 2200 Rubella and CMV IgM Control Set: Two (2) 1.5 mLPositive Control serum vials, containing human IgM antibodies toCMV; two (2) 1.5 mL Positive Control serum vials, containinghuman IgM antibodies to Rubella in a human serum matrix madefrom defibrinated plasma; and two (2) 1.5 mL Negative Controlserum vials, in a human serum matrix made from defibrinatedplasma. All antibodies are derived from human disease stateplasma. All controls contain Amikacin (0.003%), Cycloheximide(C15H23NO4) (0.009%), Amphotericin B (0.002%), CefotaximeSodium (0.002%), Ciprofloxacin (0.005%), ProClin® 300 (<0.3%),Sodium benzoate (<0.1%) and sodium azide (<0.1%).
660-0817	BioPlex 2200 Sheath Fluid: Two (2) 4 L bottles containingPhosphate Buffered Saline (PBS). ProClin® 300 (0.03%) andsodium azide (<0.1%) as preservatives.
660-0818	BioPlex 2200 Wash Solution: One (1) 10 L bottle containingPhosphate Buffered Saline (PBS) and Tween 20. ProClin® 300(0.03%) and sodium azide (<0.1%) as preservatives.
660-0000	BioPlex 2200 Instrument and Software System.

5. Intended Use

BioPlex® 2200 Rubella and CMV IgM Kit

The BioPlex® 2200 Rubella and CMV IgM kit is a multiplex flow immunoassay intended for the qualitative detection of IgM antibodies to Rubella and Cytomegalovirus (MV) in human serum and potassium EDTA or sodium heparin plasma.

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The BioPlex 2200 Rubella and CMV IgM kit is intended for use with the Bio-Rad BioPlex 2200 System.

This assay is not FDA cleared or approved for use in testing (screening) blood or plasma donors.

Performance characteristics for the Rubella and CMV IgM assays have not been evaluated in immunosupressed or organ transplant individuals. Performance characteristics of this kit in our of engall final duals. T enomance
use of peint of pare facilit use at point of care facilities.

BioPlex® 2200 Rubella and CMV IgM Calibrator Set

The BioPlex 2200 Rubella and CMV IgM Calibrator Set is intended for calibration of the BioPlex 2200 Rubella and CMV IgM Reagent Pack.

BioPlex® 2200 Rubella and CMV IgM Control Set

6. Technological Characteristics and Substantial Equivalence

The following tables summarize the similarities and differences between the BioPlex 2200 Rubella and CMV IgM kit and the predicate devices used in comparative studies with the BioPlex 2200 Rubella and CMV IgM kit.

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BioPlex® 2200 Rubella and CMV IgM Kit vs. Predicate Devices -Similarities

Item	BioPlex® 2000Rubella and CMV IgM Kit	ADVIA CentaurRubella IgM (K010668)	bioMeriéux, Inc. VIDAS®CMV IgM (K933549)
Intended Use	The BioPlex™ 2200 Rubella andCMV IgM kit is a multiplex flowimmunoassay intended for thequalitative detection of IgMantibodies to Rubella, andCytomegalovirus (CMV) in humanserum and EDTA or heparinizedplasma.The BioPlex™ 2200 Rubella andCMV IgM kit is intended for use withthe Bio-Rad BioPlex 2200 System.This kit is intended as an aid in thediagnosis of a current or recentRubella and/or CMV infection, inindividuals suspected of having oneof the respective disease statesincluding women of child bearingage.	The ADVIA Centaur andADVIA Centaur XP RubellaIgM assay is an IgM antibodycapture microparticle directchemiluminometric in vitrodiagnostic immunoassay forthe qualitative detection of IgMantibodies to the rubella virusin serum or plasma (EDTA,heparin) as an aid in thepresumptive diagnosis ofcurrent or recent infection withrubella.WARNING: The calculatedvalues for rubella IgM in agiven specimen, asdetermined by assays fromdifferent manufacturers, canvary due to differences inassay methods and reagentspecificity.	VIDAS® CMV IgM (CMVM)assay is intended for use witha VIDAS (VITEKImmunoDiagnostic AssaySystem) instrument as anautomated enzyme-linkedfluorescent immunoassay(ELFA) for the qualitativedetection of anti-CMV IgMantibodies in human serum. Itis intended to be used as anaid in the diagnosis ofcytomegalovirus infection. Itis not intended for use intesting (screening) blood orplasma donors.
Antigen	Partially purified cell lysates ofRubella, and CMV	Inactivated Rubella virus(HPV-77)	Purified and inactivated CMVantigen (Strain AD 169)
Assay Type	Qualitative detection forRubella and CMV IgM	Same	Same
Analyte Detected	Human IgM antibodiesto Rubella and CMV	Same	Same
Signal Detection	Fluorescence	Same	Same
Matrices	Serum and plasma (EDTA andheparin)	Serum and plasma (EDTAand heparin)	Same

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BioPlex® 2200 Rubella and CMV IgM Kit vs. Predicate Devices -Differences

Item	BioPlex® 2200Rubella and CMV IgM Kit	ADVIA CentaurRubella IgM (K010668)	bioMeriéux, Inc. VIDAS®CMV IgM (K933549)
Number of AnalytesSimultaneouslyDetected	Multiple (2)	Single	Single
Enzyme Conjugate	Phycoerythrin conjugated	Rubella antigen labeled withacridinium ester	Alkaline phosphataseconjugated
Matrices	Serum and plasma (EDTAand heparin)		Serum
Assay Technology	Multiplex flow immunoassay	Sandwich immunoassay usingdirect, chemiluminometrictechnology	Two-step enzymeimmunoassay sandwichmethod with fluorescentdetection (ELFA)
Signal Detection	Fluorescence	chemiluminescence
Solid Phase	Antigen-coated paramagneticmicrobead reagent.Microbeads are infused withred and infrared fluorescentdyes for bead classification.Green fluorescence from theimmunoassay label is used foranalyte measurement.	anti-human IgMFC monoclonalantibody is covalently coupled toparamagnetic particles	Antigen-coated solid phasereceptacles
Calibrator(s)	Multiple Calibrators	Single Calibrator	Single Calibrator
Controls	Negative Control and twoseparate Positive Controls forrubella and CMV IgM	Negative Control and PositiveControl specific for rubella IgM	Negative Control and PositiveControl specific for CMV IgM

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7. Performance Testing

A series of studies was conducted to evaluate the performance of the BioPlex® 2200 Rubelia and CMV IgM kit. The studies included reproducibility, interfering substances, cross-reactivity, expected values and method comparison. The results of all studies demonstrated that the BioPlex 2200 Rubella and CMV IgM kit performed according to its specifications.

A. Reproducibility

Separate internal and external reproducibility studies were conducted to evaluate the reproducibility of the BioPlex 2200 Rubella and CMV JgM kit on the BioPlex 2200 instrument. Reproducibility studies were based on the principles described in Clinical and Laboratory Standards Institute (CLSI) EP5-A2, Evaluation of Precision Performance of Quantitative Measurement Methods.

For the internal reproducibility study, three (3) panels made from serum and plasma (EDTA and heparinized) were assayed two (2) times in two (2) separate daily runs over 20 days (n=80).

The data were analyzed for within-run, between-run, between-day, and total precision and the standard deviation (SD) and percent coefficient of variation (% CV) were calculated.

The within-run precision for rubella IgM samples ranged from 4.3% to 5.5% in a serum matrix, 6.3% to 6.9% for EDTA and 4.8% to 9.5% for heparin plasma matrices. The within-run precision for CMV IgM samples range 1 from 5.3% to 9.2% in a serum matrix, 4.9% to 11.8% for EDTA and 5.2% to 12.8% for heparin plasma matrices

The external reproducibility study was performed at three (3) clinical trial sites. Three lots of reagent packs, three lots of the BioPlex 2200 Rubella and CMV IgM Calibrator Set and three lots of BioPlex 2200 Rubella and CMV IgM Control Set were evaluated. Three (3) panels made from serum and plasma (EDTA and heparinized) were tested in quadruplicate over five (5) days (4 replicates x 1 run x 5 days x 3 testing sites = 60 replicates per panel member).

The data were analyzed for within-run, between-run, between-day, between-site, and total precision and the standard deviation (SD) and percent coefficient of variation (% CV) were calculated.

The within run precision for positive samples greater than or equal to the cut-off (≥ 1.1 Al) in all sample matrices ranged from 2.7% to 13.3% for Rubella IgM and

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2.8% to 13.5% for CMV IgM. The total precision for positive samples greater than or equal to the cut-off (≥1.1 Al) ranged from 2.3% to 13.5% for Rubella India 3.0% to 14.2% for CMV IgM.

B. Interfering Substances

An interfering substances study was conducted to evaluate the potential interference of specific endogenous and exogenous substances with the BioPlex 2200 Rubella and CMV IgM assays. The study was conducted based on the principles described in Clinical and Laboratory Standards Institute (CLSI) EP7-A2, Interference Testing in Clinical Chemistry. No interference was observed with any of the substances tested. The substances and the maximum levels tested are shown in the table below.

Substance	Concentration
Hemoglobin	≤ 500 mg/dL
Bilirubin, Unconjugated	≤ 20 mg/dL
Bilirubin, Conjugated	≤ 30 mg/dL
Cholesterol	≤ 500 mg/dL
Red Blood Cells	≤ 0.4% (v/v)
Gamma Globulin	≤ 6 g/dL
Triglycerides	≤ 3300 mg/dL
Beta Carotene	≤ 0.6 mg/dL
Protein (total)	≤ 12 g/dL
Ascorbic Acid	≤ 3 mg/dL
Lithium Heparin	≤ 8000 units/dL
Sodium Heparin	≤ 8000 units/dL
EDTA	≤ 800 mg/dL
Sodium citrate	≤ 1000 mg/dL

Interfering Substances

C. Cross-Reactivity

A cross-reactivity study was performed to determine if samples from various disease states and other potentially cross-reacting agents interfere with test results when tested with the BioPlex 2200 Rubella and CMV IgM kit. Samples known to be positive for each of the potential cross-reactants, as determined by FDA cleared devices, were evaluated with the BioPlex 2200 Rubella and CMV IgM assay. All samples were pre-tested by the predicate devices and only those that tested negative by the predicate devices were further evaluated by the BioPlex 2200 Rubella and CMV IgG. The table below summarizes the number of samples scored negative by the BioPlex 2200 Rubella and CMV IgM assay within each of the crossreactant panels. No significant cross reactivity was observed except for potential

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cross reactivity of myeloma IgM samples tested with the rubella and CMV IgM assays, and EBV VCA and parvovirus B19 IgM and dsDNA samples tested with the CMV IgM assay.

PotentialCross-Reactant	BioPlex 2200 Rubella and CMV IgMResults
	N	RubellaIgM	N	CMVIgM
ANA Screen	10	10	10	9
CMV IgM	10	10	0	N/A
dsDNA (SLE clinical)	10	10	10	9ᵇ
EBV VCA IgM	10	9	8	6
hCG (pregnancy)	10	10	10	10ª
HIV IgG	10	10	10	10
HSV-1 or 2 IgM	9	8	10	10ᵇ
Hypergammaglobulinemia IgM	10	9	10	10
Influenza (compliment fixation)	10	10	10	10
Measles IgM	10	10	10	10
Mumps IgM	10	10	10	9
Myeloma IgM	9	7	7	5
Parvovirus B19 IgM	10	10ª	9	4
Rheumatoid Factor (Total)	9	9	10	9ª
Rubella IgM	0	N/A	10	10
T. gondii IgM	10	10	10	10
VZV IgM	10	10	10	10

Cross-Reactivity

a One BioPlex 2200 equivocal result

b Two BioPlex 2200 equivocal results

Note: The highlighted arcas indicate that the potential cross reactivity with either Rubella or CMV IgM could
not be ruled out not be ruled out.

D. lgM Detection

Rubella and CMV IgM-positive samples were selected and supplemented with matched specific IgG. The sample pools were scieced and supplemented with dithiothreitol (DTT) which inactivates IgM activity. The samples were assayed heat and diluted into assay range in replicates of two. Into samples were assayed head.
and OMM has measured in replicates of two. IgM was measured using Rubella and CMV IgM kit. The results are shown in below.

BioPlex 2200 Rubella and CMV IgM kit - Rubella IgM Specificity
----------------------------------------------------------------	--	--	--	--

Sample	Rubella IgM (AI)Before Treatment	DTT TreatmentAI (% recovery)
Sample 1	3.6	0.2 (5.6%)

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Sample2	3.3	0.2 (6.1%)
Sample3	3	0.2 (6.7%)
Sample4	2.7	0.2 (7.4%)
Sample5	2.6	0.2 (7.7%)
Sample6	2.5	0.2 (8%)
Sample7	2.3	0.2 (8.7%)
Sample8	2.2	0.2 (9.1%)
Sample9	2.2	0.2 (9.1%)
Sample10	2.2	0.2 (9.1%)

BioPlex 2200 Rubella and CMV IgM kit - CMV IgM Specificity

Sample	CMV IgM (AI)*Before Treatment	DTT TreatmentAI (% recovery)
Sample1	460	30.2 (6.6%)*
Sample2	341.5	26.4 (7.7%)*
Sample3	109	0.4 (0.4%)
Sample4	107	0.7 (0.7%)
Sample5	85.5	0.7 (0.8%)
Sample6	37.1	0.6 (1.6%)
Sample7	35.5	0.6 (1.7%)
Sample8	28.1	0.3 (1.1%)
Sample9	23.8	0.3 (1.3%)
Sample10	22.5	0.7 (3.1%)

*Values derived from dilution

E. Seroconversion Testing

Rubella IgM

Three Liquichek ™ Rubella IgM seroconversion panels obtained from Bio-Rad Laboratories were tested with the BioPlex 2200 Rubella and CMV IgM kit. The results shown in below were compared to a commercial method

Panel RP001	Rubella IgM
Day	Commercial MethodIndex	BioPlex 2200 Rubellaand CMV IgM kitAl
O	0.20 (Nea)	0.3 (Neg)
2	0.10 (Neg)	0.2 (Neg)
7	0.15 (Nea)	0.2 (Neg)
ರಿ	0.00 (Neg)	0.2 (Neg)
14	0.29 (Neg)	0.3 (Nea)

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17	0.91 (Eq)	1.1 (Pos)
21	9.57 (Pos)	>4.0 (Pos)
24	7.95 (Pos)	>4.0 (Pos)
28	1.87 (Pos)	1.9 (Pos)
31	4.92 (Pos)	3.7 (Pos)
35	7.31 (Pos)	3.1 (Pos)
38	3.54 (Pos)	1.8 (Pos)
42	2.43 (Pos)	1.7 (Pos)
45	1.83 (Pos)	1.3 (Pos)
50	1.95 (Pos)	1.2 (Pos)

The BioPlex 2200 Rubella and CMV IgM test detected Rubella IgM four days earlier demonstrating greater sensitivity.

Panel RP011	Rubella IgM
Day	Commercial Method	BioPlex 2200 Rubella
	Index	and CMV IgM kit
		AI
0	0.21 (Neg)	0.2 (Neg)
3	0.00 (Neg)	0.2 (Neg)
9	0.02 (Neg)	0.2 (Neg)
12	0.00 (Neg)	0.2 (Neg)
16	1.64 (Pos)	1.1 (Pos)
19	5.94 (Pos)	>4.0 (Pos)
24	8.95 (Pos)	>4.0 (Pos)
27	7.69 (Pos)	>4.0 (Pos)
31	4.50 (Pos)	3.7 (Pos)
36	2.70 (Pos)	2.4 (Pos)
39	1.78 (Pos)	1.8 (Pos)
43	1.12 (Pos)	1.2 (Pos)
46	0.77 (Neg)	0.8 (Neg)
50	0.43 (Neg)	0.8 (Neg)
53	0.32 (Neg)	0.6 (Neg)
57	0.32 (Neg)	0.6 (Neg)
60	0.13 (Neg)	0.4 (Neg)
64	0.37 (Neg)	0.5 (Neg)
67	0.00 (Neg)	0.4 (Neg)
71	0.00 (Neg)	0.5 (Neg)

The BioPlex 2200 Rubella and CMV IgM test showed comparable performance to a commercial method.

Panel RP014	Rubella IgM
Day	Commercial MethodIndex	BioPlex 2200 Rubellaand CMV IgM kitAI
0	0.00 (Neg)	<0.2 (Neg)
5	0.00 (Neg)	<0.2 (Neg)
7	0.00 (Neg)	<0.2 (Neg)

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12	0.00 (Neg)	<0.2 (Neg)
14	0.23 (Neg)	0.7 (Neg)
19	2.44 (Pos)	2.3 (Pos)
21	2.55 (Pos)	2.4 (Pos)
26	2.58 (Pos)	2.2 (Pos)
28	2.15 (Pos)	1.8 (Pos)
33	1.5 (Pos)	1.6 (Pos)
35	1.5 (Pos)	1.2 (Pos)
40	1.00 (Pos)	1.0 (Eq)
42	0.76 (Neg)	0.8 (Neg)

The BioPlex 2200 Rubella and CMV IgM test showed comparable performance to a commercial method, except for the day 40 sample which scored equivocal in the BioPlex 2200 Rubella and CMV IgM test and positive (at the cutoff) in the comparison test.

CMV IgM

Bio-Rad Laboratories Liquichek™ CMV IgM Seroconversion panel was assayed with BioPlex 2200 Rubella and CMV IgM kits and a commercial method. The results are shown in below. The BioPlex 2200 Rubella and CMV IgM test was able to detect CMV IgM in the Libr Ion LEGO Rubella and CMV IgM test was able equivocal at 59 days.

Panel RP003	CMV IgM
Day	Commercial MethosIndex	BioPlex 2200 Rubellaand CMV IgM kitAI
1	1.16 (Pos)	>4.0 (Pos)
4	1.62 (Pos)	>4.0 (Pos)
8	2.44 (Pos)	>4.0 (Pos)
51	1.16 (Pos)	3.4 (Pos)
55	1.04 (Pos)	3.0 (Pos)
59	0.84 (Eq)	2.1 (Pos)
65	0.85 (Eq)	2.3 (Pos)
67	0.75 (Eq)	2.0 (Pos)
72	0.68 (Neg)	1.7 (Pos)
74	0.62 (Neg)	1.5 (Pos)
79	0.64 (Neg)	1.8 (Pos)
84	0.54 (Neg)	1.7 (Pos)
88	0.72 (Eq)	2.1 (Pos)
95	0.59 (Neg)	1.7 (Pos)
99	0.65 (Neg)	1.6 (Pos)

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ட் Expected Values

The observed prevalence for the Rubella and CMV IgM using the BioPlex 2200 Rubella and CMV IgM assay was determined using samples submitted for Rubella (300, US) or CMV IgM (400, 300 US +100 Europe) testing. The results are presented in the tables below. The predictive values of the test are dependent on the prevalence. As rubella incidence decreases, the predicative positive value of rubella igM results decreases.

Note: Each laboratory should establish frequency distributions for their specific patient populations.

Age	Gender	Rubella IgM(US)		CMV IgM(US)		CMV IgM(EU)
		Pos/Tota	% revalence Trevalence Station	os/Tota	revalenc%	Pos/Tota	Prevalen%
0 - 10	F	0/13	0.0%	0/4	0.0%	0/0	N/A
	M	0/1 1	0.0%	Ole	0.0%	0/2	0.0%
11 - 20	F	0/44	0.0%	1/19	5.3%	0/4	0.0%
	M	0/12	0.0%	0/20	0.0%	0/1	0.0%
21 - 30	F	4/80	5.0%	1/41	2.4%	1/36	2.8%
	M	018	0.0%	0/10	0.0%	0/2	0.0%
31 - 40	F	4/55	7.3%	1/34	2.9%	3/42	7.1%
	M	0/8	0.0%	3/17	17.6%	0/1	0.0%
41 - 50	F	3/26	11.5%	1/35	2.9%	0/6	0.0%
	M	1/12	8.3%	0/19	0.0%	0/2	0.0%
51 - 60	F	0/19	0.0%	1/23	4.3%	0/1	0.0%
	M	0/0	0.0%	1/33	0.0%	0/1	0.0%
61 - 70	F	0/1	0.0%	0/16	0.0%	0/1	0.0%
	M	0/1	0.0%	0/14	0.0%	0/0	N/A
71 +	F	0/0	N/A	0/2	0.0%	0/0	N/A
	M	0/1	0.0%	077	0.0%	0/1	0.0%
	Total		4.0%**	9/300	3.0%	4/100	4.0%

Expected values using the BioPlex 2200 Rubella and CMV IgM kit in test ordered population

*One of pregnant women (N=71) was Rubella IgM positive.
**Since 2001 The ingidonea of Dubelle

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Method Comparison G.

Performance of the BioPlex 2200 Rubella and CMV IgM kit was evaluated against corresponding commercially available Rubella and CMV kits. Three clinical sites tested 700 prospective samples submitted for: Rubella (300 - U.S.), and CMV IgM testing (400,300 U.S. + 100 Europe). Of the 300 samples submitted for Rubella IgM testing, 71 females were pregnant women. Results are shown in the table below:

Of the 300 clinical ordered samples test for Rubella IgM, the positive percent agreement was 66.7% and the negative percent agreement was 95.6%.

Of the 71 clinical ordered pregnancy samples test for Rubella IgM, the negative percent agreement was 96.8%.

Of the 400 clinical ordered samples tested for CMV IgM, the positive percent agreement was 53.8% and the negative percent agreement was 97.7%.

The results are presented in the table below.

Test Ordered		BioPlex 2200 Rubella and CMV IgM kit
Rubella IgM	Test Ordered	Pos (+)	Eqv	Neg (-)	Total	Pos (+)% Agreement	Neg (-)% Agreement
	Pos (+)	2	0	1	3	66.7%(2/3)95% CI20.8 - 93.9%	95.6%(282/295)95% CI92.6 - 97.4%
	Eqv	0	2	0	2
	Neg (-)	10	3	282	295
	Total	12	5	283	300
Pregnant Women*	Pos (+)	0	0	0	0	N/A	96.8%(70/71)95%CI92.8% - 99.8%
	Eqv	0	0	0	0
	Neg (-)	1	0	70	71
	Total	1	0	70	71
	Pos (+)	7a	0	3a	10	53.8%(7/13)	97.7%(377/386)

Method Comparison: Prospective Study

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Eqv	1	1	3	5
Neg (-)	5b	3	377a	385
Total	13	4	383	400

a. One sample that was equivocal by the predicate device was adjudicated by two out of three FDA cleared devices.

b. Two samples that were equivocal by the predicated by two out of three HDA cleared devices

Pregnant women is the subset of the test ordered population.

Comparative Testing: Retrospective Study

Performance of the Rubella and CMV IgM kit was evaluated against corresponding commercially available Rubella and CMV IgM immunoassays. Three clinical sites tested 107 Rubella (45 females with 89% in 15-45 age group, 49 males and 13 with unknown gender) and 229 CMV (144 females with 84% in 15-45 age group and 85 males) presumptive IgM positive samples. Positive samples for Rubella and CMV IgM were selected by another FDA cleared test and the respective commercially available immunoassays used for the comparative analysis. The results are presented in the table below.

Presumptive Positive		BioPlex Rubella and CMV IgM kit
for Rubella or CMVIgM		Pos(+)	Eqv	Neg(-)	Total	Pos(+)% Agreement
Commercially Available Immunoassay	Rubella IgM	Pos(+)	103	1	3	107	96.3%
		Eqv	0	0	0	0	(103/107)
		Neg(-)	0	0	0	0	95% CI
		Total	103	1	3	107	90.3 – 98.5%
	CMV IgM	Pos(+)	209a	3	17b	229	91.3%
		Eqv	0	0	0	0	(209/229)
		Neg(-)	0	0	0	0	95% CI
		Total	209	3	17	229	86.9 - 94.3%

Characteristics of Samples with Presumptive Positive Status

e that was equivocal by the predicate device was adjudicated by two out of three FDA cleared devices.

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Matrix Comparison

Matched serum and plasma (potassium EDTA and sodium heparin) samples drawn from 20 individual donors were acquired from commercial sources. All samples were evaluated in replicates of 10. Mean plasma Al values were compared to matched mean serum Al values. Scatter plots comparing the performance of serum samples against potassium EDTA and sodium heparin plasma samples along with the corresponding slopes of regression and correlation coefficient (r) are shown in below. All assays pass the slope specification and correlation
specification of + 0.2, and correction as the slope specification of 1.0 ± 0.2, in specification of ± 0.2, and correlation coefficient (r) of ≥ 0.98.

Image /page/15/Figure/3 description: The image shows a scatter plot titled "Matrix Comparison Mean Test vs Mean Comparison". The x-axis is labeled "Serum (Al)" and ranges from 0 to 2.5. The y-axis is labeled "EDTA Plasma (Al)" and ranges from 0 to 2.5. A linear regression line is plotted through the data points, with the equation y = 1.0369x - 0.0077 and an r-value of 0.9971.

Image /page/15/Figure/4 description: The image is titled "Figure 1. Rubella IgM: EDTA vs. Serum (N=20)". The figure is comparing Rubella IgM levels in EDTA versus serum samples. The sample size is N=20.

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

Figure 2. Rubella lgM: Heparin vs. Serum (N=20)

Image /page/16/Figure/1 description: This image is a scatter plot titled "Matrix Comparison Mean Test vs Mean Comparison". The x-axis is labeled "Serum (Al)" and ranges from 0 to 2.5. The y-axis is labeled "Heparin Plasma (Al)" and ranges from 0 to 2.5. A line of best fit is plotted on the scatter plot, and the equation for the line is y = 1.0219x - 0.0123, with an r value of 0.9976.

Figure 3. CMV IgM: EDTA vs. Serum (N=20)

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

Image /page/17/Figure/0 description: This image is a scatter plot titled "Matrix Comparison Mean Test vs Mean Comparison". The x-axis is labeled "Serum (Al)" and ranges from 0 to 2, while the y-axis is labeled "Heparin Plasma-(Al)" and also ranges from 0 to 2. The plot shows a strong positive correlation between the two variables, with data points clustered closely around a straight line, and the equation of the line is y = 1.0000x - 0.0000, and the r value is 0.9945.

Figure 4. CMV IgM: Heparin vs. Serum (N=20)

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

Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993

Bio- Rad Laboratories, Inc. BioPlex 2200 Division c/o Juang Wang Regulatory Affairs Representative 5500 East Second Street Benicia, CA 94510

DEC - 3 2010

Re: K092587

Trade/Device Name: BioPlex® 2200 Rubella and CMV IgM Kit on the BioPlex® 2200 Multi Analyte Detection System Regulation Number: 21CFR§866.3510 Regulation Name: Rubella Virus Serological Reagents Regulatory Class: Class II Product Code: LFX, LKQ, LFZ, JIX, JJX Dated: November 30, 2010 Received: December 1, 2010

Dear Mr. Wang:

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, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into class II (Special Controls), 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); medical device reporting (reporting of

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

Page 2 - Juang Wang

medical device-related adverse events) (21 CFR 803); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820). 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 advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office

of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Sma!l Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely vours.

Jouga Hoynes

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

Enclosure

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

Indication(s) for use

DEC - 3 2010

510(k) Number (if known): K092587

Device Name:BioPlex® 2200 Rubella and CMV IgM Kit on the BioPlex® 2200 Multi Analyte

Detection System

BioPlex® 2200 Rubella and CMV IgM Calibrator Set BioPlex® 2200 Rubella and CMV IgM Control Set

Indications for Use:

The BioPlex® 2200 Rubella and CMV IgM kit

The BioPlex 2200 Rubella and CMV IgM kit is intended for use with the Bio-Rad BioPlex 2200 System.

This assay is not FDA cleared or approved for use in testing (screening) blood or plasma donors.

BioPlex® 2200 Rubella and CMV IgM Calibrator Set

The BioPlex® 2200 Rubella and CMV IgM Calibrator Set is intended for the calibration of the BioPlex® 2200 Rubella and CMV IgM Reagent Pack.

BioPlex® 2200 Rubella and CMV IgM Control Set

The BioPlex® 2200 Rubella and CMV IgM Control Set is intended for use as an assayed quality control to monitor the overall performance of the BioPlex 2200 instrument and BioPlex 2200 Rubella and CMV IgM Reagent Pack in the clinical laboratory. The performance of the BioPlex 2200 Rubella and CMV IgM Control Set has not been established with any other Rubella and CMV IgM assays.

Over-the-Counter Use Prescription Use X AND/OR (21 CFR 801 Subpart C) (Part 21 CFR 801 Subpart D)

(Please do not write below this line-Continue on another page if needed)

Concurrence of CDRH, Office of In Vitro Diagnostic Device Evaluation and Safety (OVD)

Une Schif

Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K092587

Regulation Number and Section

§ 866.3510 Rubella virus serological reagents.

(a)
Identification. Rubella virus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to rubella virus in serum. The identification aids in the diagnosis of rubella (German measles) or confirmation of a person's immune status from past infections or immunizations and provides epidemiological information on German measles. Newborns infected in the uterus with rubella virus may be born with multiple congenital defects (rubella syndrome).(b)
Classification. Class II. The special controls for this device are:(1) National Committee for Clinical Laboratory Standards':
(i) 1/LA6 “Detection and Quantitation of Rubella IgG Antibody: Evaluation and Performance Criteria for Multiple Component Test Products, Speciment Handling, and Use of the Test Products in the Clinical Laboratory, October 1997,”
(ii) 1/LA18 “Specifications for Immunological Testing for Infectious Diseases, December 1994,”
(iii) D13 “Agglutination Characteristics, Methodology, Limitations, and Clinical Validation, October 1993,”
(iv) EP5 “Evaluation of Precision Performance of Clinical Chemistry Devices, February 1999,” and
(v) EP10 “Preliminary Evaluation of the Linearity of Quantitive Clinical Laboratory Methods, May 1998,”
(2) Centers for Disease Control's:
(i) Low Titer Rubella Standard,
(ii) Reference Panel of Well Characterized Rubella Sera, and
(3) World Health Organization's International Rubella Standard.