← Product Code [PUQ](/submissions/MI/subpart-d%E2%80%94serological-reagents/PUQ) · K170509

# BioPlex 2200 ToRC IgM, BioPlex 2200 ToRC IgM Calibrator Set, BioPlex 2200 ToRC IgM Control Set (K170509)

_Bio-Rad Laboratories · PUQ · May 19, 2017 · Microbiology · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/PUQ/K170509

## Device Facts

- **Applicant:** Bio-Rad Laboratories
- **Product Code:** [PUQ](/submissions/MI/subpart-d%E2%80%94serological-reagents/PUQ.md)
- **Decision Date:** May 19, 2017
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 866.3510
- **Device Class:** Class 2
- **Review Panel:** Microbiology

## Indications for Use

The BioPlex 2200 ToRC IgM kit is a multiplex flow immunoassay intended for the qualitative detection of IgM antibodies to Toxoplasma gondii (T. gondii), Rubella and Cytomegalovirus (CMV) in human serum and plasma (K3 EDTA, lithium heparin, or sodium heparin). The BioPlex 2200 ToRC 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 T. gondii, 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 ToRC IgM assay have not been evaluated in immunosuppressed 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 ToRC IgM Calibrator Set is intended for the calibration of the BioPlex 2200 ToRC IgM Reagent Pack. The BioPlex 2200 ToRC 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 ToRC IgM Reagent Pack in the clinical laboratory.

## Device Story

Multiplex flow immunoassay for qualitative detection of IgM antibodies to T. gondii, Rubella, and CMV. Input: human serum or plasma samples. Process: automated two-step assay on BioPlex 2200 System; patient sample mixed with antigen-coated fluoromagnetic beads; immobilized IgM identified via fluorescent anti-human IgM reporter conjugate. Output: relative fluorescent intensity (RFI) converted to Antibody Index (AI). Used in clinical laboratories by trained personnel. Results aid clinicians in diagnosing current or recent infections. Benefits include simultaneous multi-analyte testing from a single sample, reducing sample volume requirements and improving laboratory efficiency.

## Clinical Evidence

Clinical performance evaluated via prospective study (N=2,129) and retrospective study (N=524) across three U.S. sites. Prospective study compared results against predicate immunoassays, showing high negative agreement (97.4%–100%) and variable positive agreement. Retrospective study of presumptive positive samples showed positive agreement of 97.1% (T. gondii), 98.0% (Rubella), and 98.5% (CMV). CDC reference panel testing for T. gondii IgM showed 100% positive and negative agreement. Specificity confirmed via DTT treatment to inactivate IgM. Bench testing included precision (CLSI EP5-A3), interference (CLSI EP7-A2), and matrix comparison (CLSI EP09-A3).

## Technological Characteristics

Multiplexed flow immunoassay using antigen-coated paramagnetic microbeads. Detection via dual-laser fluorescence. Reagents include phycoerythrin-conjugated anti-human IgM reporter. Sample matrix: serum, K3 EDTA, lithium heparin, sodium heparin. System: BioPlex 2200. Software-controlled automated processing. Preservatives: ProClin 300, sodium benzoate, sodium azide.

## Regulatory 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).

## Special Controls

*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.

## Predicate Devices

- BioPlex 2200 Rubella and CMV IgM Kit ([K092587](/device/K092587.md))
- bioMeriéux, Inc. VIDAS® TOXO IgM ([K923166](/device/K923166.md))

## Submission Summary (Full Text)

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>
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number:
K170509

B. Purpose for Submission:
New Device

C. Measurand:
IgM antibodies to *Toxoplasma gondii* (T. gondii), Rubella virus and Cytomegalovirus (CMV)

D. Type of Test:
Multiplexed Microparticle Immunoassay (multiplexed fluoromagnetic bead assay)

E. Applicant:
Bio-Rad Laboratories

F. Proprietary and Established Names:
BioPlex 2200 ToRC IgM
BioPlex 2200 ToRC IgM Calibrator Set
BioPlex 2200 ToRC IgM Control Set

G. Regulatory Information:

|  Product code | Classification | Regulation section | Panel  |
| --- | --- | --- | --- |
|  PUQ: Multiplex flow immunoassay, T. gondii, Rubella, CMV IgM | Class II | 866.3510 - Rubella Virus Serological Reagents | Microbiology  |
|  JIX: Calibrator, multi- analyte mixture | Class II | 862.1150 - Calibrator | Clinical Chemistry  |
|  JJX: Single (specified) analyte controls (assayed and unassayed) | Class I | 862.1660 - Quality Control Material (assayed and unassayed) | Clinical Chemistry  |

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Note: The BioPlex™ 2200 ToRC IgM Kit is a multiplex immunoassay for the detection of IgM antibodies to *T. gondii*, Rubella virus and CMV. This device is classified as Class II as described above and a new product code is assigned for this device as the primary product code, listed under the regulation section for Rubella reagents. The following are the additional regulation sections and product codes that are applicable to the other analytes detected by the subject device of this submission.

866.3175 - Cytomegalovirus serological reagents. Class II (Microbiology). Antibody IgM, IF, cytomegalovirus virus (LKQ) and Enzyme Linked Immunosorbent Assay, Cytomegalovirus (LFZ)

866.3780 – *Toxoplasma gondii* Serological Agents. Class II (Microbiology). Enzyme Linked Immunosorbent Assay, *Toxoplasma gondii* (LGD)

H. Intended Use:

1. Intended use(s):

**BioPlex 2200 ToRC IgM kit**

The BioPlex 2200 ToRC IgM kit is a multiplex flow immunoassay intended for the qualitative detection of IgM antibodies to *Toxoplasma gondii* (*T. gondii*), Rubella and Cytomegalovirus (CMV) in human serum and plasma (K3 EDTA, lithium heparin, or sodium heparin).

The BioPlex 2200 ToRC 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 *T. gondii*, 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 ToRC IgM assays have not been evaluated in immunosuppressed 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.

**BioPlex 2200 ToRC IgM Calibrator Set**

The BioPlex 2200 ToRC IgM Calibrator Set is intended for the calibration of the BioPlex 2200 ToRC IgM Reagent Pack.

**BioPlex 2200 ToRC IgM Control Set**

The BioPlex 2200 ToRC 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 ToRC IgM Reagent Pack in the clinical laboratory.

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2. Indication(s) for use:
Same as Intended Use

3. Special conditions for use statement(s):
For Prescription use only

4. Special instrument requirements:
Bio-Rad BioPlex 2200 System

I. Device Description:
BioPlex ToRC IgM Reagent Pack:

The BioPlex ToRC IgM Reagent Pack includes the following components:

- One (1) 10 mL vial, containing dyed beads coated with lysates of T. gondii, Rubella virus and CMV plus an Internal Standard bead (ISB) and a Serum Verification bead (SVB) in buffer with Glycerol and protein stabilizers (bovine and caprine). ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) as preservatives.
- One (1) 5 mL vial, containing phycoerythrin-conjugated murine monoclonal anti-human IgM antibody and phycoerythrin-conjugated murine monoclonal anti-human FXIII antibody, in buffer with protein stabilizers (bovine and murine). ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) as preservatives.
- One (1) 10 mL vial, containing goat anti-human IgG antibody and protein stabilizers (bovine and murine) in buffer. ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) as preservatives.

BioPlex 2200 ToRC IgM Calibrator:

The BioPlex 2200 ToRC IgM Calibrator set contains two (2) 0.5 mL vials. The calibrators are provided in a human serum matrix made from defibrinated plasma with added known analyte concentrations consisting of HuCAL recombinant IgM antibodies against Rubella virus and human disease state plasma derived antibodies against T. gondii and CMV. All calibrators contain ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) as preservatives.

BioPlex 2200 ToRC IgM Control:

The BioPlex 2200 ToRC IgM Control set contains two (2) 1.5 mL Positive Control serum vials, containing human disease state plasma derived IgM antibodies against T. gondii and CMV, and HuCAL® recombinant IgM antibodies against Rubella virus in a human serum matrix made from defibrinated plasma; and two (2) 1.5 mL Negative Control serum vials, in a human serum matrix made from defibrinated plasma. All controls contain Amikacin (0.003%), Cycloheximide (C15H23NO4) (0.009%), Amphotericin B (0.002%), Cefotaxime Sodium (0.002%), Ciprofloxacin (0.005%), ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%).

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J. Substantial Equivalence Information:

1. Predicate device name(s):
BioPlex 2200 Rubella and CMV IgM Kit
bioMérieux, Inc. VIDAS® TOXO IgM

2. Predicate 510(k) number(s):
K092587
K923166

3. Comparison with predicate:
BioPlex 2200 Rubella and CMV IgM Kit

|  Similarities  |   |   |
| --- | --- | --- |
|  Item | Device
BioPlex 2200 ToRC IgM | Predicate
BioPlex 2200 Rubella and CMV IgM Kit K092587)  |
|  Intended use | Qualitative detection of IgM antibodies to Toxoplasma gondii (T. gondii), Rubella and Cytomegalovirus (CMV)
To be used as an aid in the diagnosis of a current or recent T. gondii, Rubella and/or CMV infection | Qualitative detection of IgM antibodies to Rubella and Cytomegalovirus (CMV)
To be used as an aid in the diagnosis of a current or recent, Rubella and/or CMV infection  |
|  Technology | Automated multiplex flow immunoassay | same  |
|  Differences  |   |   |
| --- | --- | --- |
|  Item | Device
BioPlex 2200 ToRC IgM | Predicate
BioPlex 2200 Rubella and CMV IgM Kit K092587)  |
|  Matrix | Serum and Plasma (K3 EDTA and Lithium or Sodium Heparin) | Serum, potassium EDTA or Sodium Heparin plasma  |

VIDAS® TOXO IgM

|  Similarities  |   |   |
| --- | --- | --- |
|  Item | Device
BioPlex 2200 ToRC IgM | Predicate
bioMérieux, Inc., VIDAS® TOXO IgM K923166  |
|  Intended use | Qualitative detection of IgM antibodies to Toxoplasma gondii (T. gondii), Rubella | Qualitative detection of anti-Toxoplasma gondii antibodies  |

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|  Similarities  |   |   |
| --- | --- | --- |
|  Item | Device BioPlex 2200 ToRC IgM | Predicate bioMérieux, Inc., VIDAS® TOXO IgM K923166  |
|   | and Cytomegalovirus (CMV) To be used as an aid in the diagnosis of a current or recent T. gondii, Rubella and/or CMV infection | To be used as an aid in the diagnosis of acute, recent, or reactivated Toxoplasma gondii infection  |
|  Differences  |   |   |
| --- | --- | --- |
|  Item | Device BioPlex 2200 ToRC IgM | Predicate bioMérieux, Inc., VIDAS® TOXO IgM K923166  |
|  Technology | Automated multiplex flow immunoassay | Manual enzyme linked fluorescent immunoassay (ELFA)  |
|  Matrix | Serum and Plasma (K3 EDTA and Lithium or Sodium Heparin) | Serum  |

# K. Standard/Guidance Document Referenced (if applicable):

EP05-A3, Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline, Third Edition (Vol. 34 No.13)

EP07-A2, Interference Testing in Clinical Chemistry, Approved Guideline, Second Edition (Vol. 25 No.27)

EP09-A3, Measurement Procedure Comparison and Bias Estimation Using Patient Samples, Approved Guideline- Third Edition (Vol. 33, No. 11)

EP12-A2, User Protocol for Evaluation of Qualitative Test Performance; Approved Guideline-Second Edition (Vol. 28, No. 3)

EP15-A3, User Verification of Precision and Estimation of Bias; Approved Guideline - Third Edition (Vol. 34, No. 12)

EP25-A, Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline (Vol. 29, No. 20)

# L. Test Principle:

The BioPlex 2200 ToRC IgM kit employs a panel of three antigen-coated fluoromagnetic beads with unique fluorescent signatures to identify the presence of IgM class antibodies to  $T.$  gondii, Rubella virus, and CMV antigens in a two-step assay format. In the first step, the system combines an aliquot of patient sample with sample diluent and bead reagent and then agitates the mixture at  $37^{\circ}\mathrm{C}$ . In the second step, immobilized IgM is identified indirectly using a fluorescent

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anti-human IgM reporter conjugate in a manner similar to antibody detection using an enzyme-linked reporter in an EIA. The assay is calibrated using a set of two distinct calibrator vials, supplied separately by Bio-Rad Laboratories. One vial containing negative sample, one vial containing T. gondii IgM, Rubella IgM, and CMV IgM are used for qualitative calibration of the assays. Two control beads are used. One is used to normalize assay output for fluctuations in detector function (internal standard bead, ISB) and the other control bead is used to verify that the sample is serum or plasma (serum verification bead, SVB). The fluorescent properties of the beads allow multi-analyte data to be acquired simultaneously from a single sample and segregated based upon the fluorescent codes embedded in the antigen-coated and control beads. The magnetic properties of the beads allow rapid washing to remove unbound molecules in between assay steps. Bead classification and reporter data are acquired as the beads flow through the reader which uses a dual laser detector employing the same principles utilized by fluorescence activated cell sorters. Raw data are reported as relative fluorescent intensity (RFI).

## M. Performance Characteristics (if/when applicable):

### 1. Analytical performance:

#### a. Precision/Reproducibility:

**Precision:**

Precision testing of the BioPlex 2200 ToRC IgM kit on the BioPlex 2200 instrument was performed in accordance with the CLSI EP5-A3 guideline. Samples were tested in duplicate, two (2) runs per day, over 20 days (2 replicates per run x 2 runs per day x 20 days (N= 80 data points per panel member) using one reagent lot, calibrator set and control set. The data were analyzed for within-run (repeatability), between-run, between-day, and total reproducibility and the mean (AI), standard deviation (AI) and percent coefficient of variation (%CV) are summarized in the tables below.

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|  BioPlex 2200 Toxo IgM |   |   |   | Within-Run |   | Between-Run |   | Between-Day |   | Total  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  Sample Type | T. gondii IgM Panel Members | N | Mean (AI) | SD | %CV | SD | %CV | SD | %CV | SD | %CV  |
|  Serum | Negative | 80 | 0.5 | 0.039 | 7.9% | 0.000 | 0.0% | 0.007 | 1.3% | 0.039 | 8.0%  |
|   |  High Negative | 80 | 0.7 | 0.034 | 4.8% | 0.011 | 1.6% | 0.012 | 1.7% | 0.037 | 5.3%  |
|   |  Cut-Off | 80 | 0.9 | 0.050 | 5.5% | 0.000 | 0.0% | 0.031 | 3.4% | 0.059 | 6.5%  |
|   |  Cut-Off | 80 | 1.0 | 0.064 | 6.3% | 0.000 | 0.0% | 0.030 | 2.9% | 0.071 | 6.9%  |
|   |  Low Positive | 80 | 1.3 | 0.057 | 4.3% | 0.042 | 3.2% | 0.039 | 2.9% | 0.081 | 6.1%  |
|   |  Positive | 80 | 2.3 | 0.113 | 5.0% | 0.000 | 0.0% | 0.034 | 1.5% | 0.118 | 5.2%  |
|   |  Positive | 80 | 3.3 | 0.164 | 4.9% | 0.034 | 1.0% | 0.053 | 1.6% | 0.175 | 5.3%  |
|  Potassium EDTA | Negative | 80 | 0.5 | 0.019 | 3.9% | 0.000 | 0.0% | 0.002 | 0.4% | 0.019 | 3.9%  |
|   |  High Negative | 80 | 0.7 | 0.046 | 6.2% | 0.000 | 0.0% | 0.027 | 3.7% | 0.053 | 7.2%  |
|   |  Cut-Off | 80 | 0.9 | 0.056 | 6.3% | 0.000 | 0.0% | 0.017 | 1.9% | 0.059 | 6.6%  |
|   |  Low Positive | 80 | 1.2 | 0.073 | 6.0% | 0.000 | 0.0% | 0.049 | 4.0% | 0.088 | 7.3%  |
|   |  Positive | 80 | 2.0 | 0.089 | 4.4% | 0.037 | 1.8% | 0.062 | 3.1% | 0.114 | 5.7%  |
|   |  Positive | 80 | 3.2 | 0.167 | 5.3% | 0.059 | 1.9% | 0.098 | 3.1% | 0.203 | 6.4%  |
|  Sodium Heparin | Negative | 80 | 0.4 | 0.045 | 10.3% | 0.000 | 0.0% | 0.018 | 4.1% | 0.048 | 11.1%  |
|   |  High Negative | 80 | 0.7 | 0.032 | 4.3% | 0.030 | 4.0% | 0.025 | 3.4% | 0.050 | 6.8%  |
|   |  Cut-Off | 80 | 0.9 | 0.058 | 6.2% | 0.025 | 2.7% | 0.022 | 2.4% | 0.067 | 7.2%  |
|   |  Low Positive | 80 | 1.3 | 0.068 | 5.3% | 0.000 | 0.0% | 0.035 | 2.7% | 0.076 | 5.9%  |
|   |  Positive | 80 | 2.1 | 0.109 | 5.3% | 0.000 | 0.0% | 0.060 | 2.9% | 0.124 | 6.0%  |
|   |  Positive | 80 | 3.1 | 0.117 | 3.8% | 0.000 | 0.0% | 0.049 | 1.6% | 0.127 | 4.1%  |
|  Lithium Heparin | Negative | 80 | 0.5 | 0.050 | 10.9% | 0.011 | 2.4% | 0.010 | 2.3% | 0.052 | 11.4%  |
|   |  High Negative | 80 | 0.7 | 0.042 | 6.0% | 0.000 | 0.0% | 0.017 | 2.4% | 0.045 | 6.4%  |
|   |  Cut-Off | 80 | 0.9 | 0.061 | 6.7% | 0.000 | 0.0% | 0.018 | 2.0% | 0.064 | 7.0%  |
|   |  Low Positive | 80 | 1.3 | 0.063 | 5.0% | 0.050 | 4.0% | 0.019 | 1.5% | 0.083 | 6.6%  |
|   |  Positive | 80 | 2.0 | 0.108 | 5.4% | 0.045 | 2.2% | 0.031 | 1.6% | 0.121 | 6.1%  |
|   |  Positive | 80 | 3.1 | 0.145 | 4.7% | 0.000 | 0.0% | 0.064 | 2.1% | 0.159 | 5.1%  |

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|  BioPlex 2200 Rubella IgM |   |   |   | Within-Run |   | Between-Run |   | Between-Day |   | Total  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  Sample Type | Rubella IgM Panel Members | N | Mean (AI) | SD | %CV | SD | %CV | SD | %CV | SD | %CV  |
|  Serum | Negative | 80 | 0.3 | 0.011 | 3.7% | 0.000 | 0.0% | 0.000 | 0.0% | 0.011 | 3.7%  |
|   |  High Negative | 80 | 0.6 | 0.040 | 6.6% | 0.000 | 0.0% | 0.016 | 2.6% | 0.043 | 7.1%  |
|   |  Cut-Off | 80 | 0.8 | 0.049 | 5.9% | 0.000 | 0.0% | 0.014 | 1.6% | 0.051 | 6.1%  |
|   |  Cut-Off | 80 | 1.1 | 0.066 | 5.8% | 0.000 | 0.0% | 0.031 | 2.7% | 0.073 | 6.4%  |
|   |  Low Positive | 80 | 1.2 | 0.063 | 5.4% | 0.027 | 2.3% | 0.027 | 2.2% | 0.074 | 6.3%  |
|   |  Positive | 80 | 1.5 | 0.085 | 5.7% | 0.039 | 2.6% | 0.000 | 0.0% | 0.094 | 6.2%  |
|   |  Positive | 80 | 2.7 | 0.146 | 5.5% | 0.000 | 0.0% | 0.000 | 0.0% | 0.146 | 5.5%  |
|   |  Positive | 80 | 3.0 | 0.157 | 5.3% | 0.035 | 1.2% | 0.091 | 3.0% | 0.185 | 6.2%  |
|  Potassium EDTA | Negative | 80 | 0.3 | 0.011 | 3.7% | 0.000 | 0.0% | 0.000 | 0.0% | 0.011 | 3.7%  |
|   |  High Negative | 80 | 0.6 | 0.045 | 7.0% | 0.000 | 0.0% | 0.029 | 4.6% | 0.053 | 8.4%  |
|   |  Cut-Off | 80 | 1.0 | 0.055 | 5.6% | 0.016 | 1.6% | 0.026 | 2.7% | 0.063 | 6.5%  |
|   |  Low Positive | 80 | 1.1 | 0.064 | 5.7% | 0.016 | 1.4% | 0.016 | 1.4% | 0.068 | 6.1%  |
|   |  Positive | 80 | 1.6 | 0.084 | 5.4% | 0.000 | 0.0% | 0.057 | 3.7% | 0.101 | 6.5%  |
|   |  Positive | 80 | 2.4 | 0.118 | 4.9% | 0.000 | 0.0% | 0.084 | 3.5% | 0.145 | 6.0%  |
|   |  Positive | 80 | 3.0 | 0.168 | 5.6% | 0.000 | 0.0% | 0.103 | 3.4% | 0.198 | 6.5%  |
|  Sodium Heparin | Negative | 80 | 0.3 | 0.045 | 13.3% | 0.011 | 3.3% | 0.014 | 4.1% | 0.048 | 14.4%  |
|   |  High Negative | 80 | 0.6 | 0.042 | 6.8% | 0.011 | 1.8% | 0.023 | 3.7% | 0.049 | 8.0%  |
|   |  Cut-Off | 80 | 1.0 | 0.045 | 4.6% | 0.034 | 3.4% | 0.029 | 2.9% | 0.063 | 6.4%  |
|   |  Low Positive | 80 | 1.1 | 0.066 | 5.8% | 0.016 | 1.4% | 0.018 | 1.6% | 0.070 | 6.2%  |
|   |  Positive | 80 | 1.6 | 0.092 | 5.7% | 0.000 | 0.0% | 0.044 | 2.7% | 0.102 | 6.3%  |
|   |  Positive | 80 | 2.5 | 0.123 | 4.9% | 0.039 | 1.5% | 0.036 | 1.4% | 0.134 | 5.3%  |
|   |  Positive | 80 | 3.2 | 0.155 | 4.9% | 0.019 | 0.6% | 0.029 | 0.9% | 0.159 | 5.0%  |
|  Lithium Heparin | Negative | 80 | 0.3 | 0.019 | 6.5% | 0.000 | 0.0% | 0.000 | 0.0% | 0.019 | 6.5%  |
|   |  High Negative | 80 | 0.6 | 0.047 | 8.3% | 0.000 | 0.0% | 0.023 | 4.1% | 0.053 | 9.2%  |
|   |  Cut-Off | 80 | 1.0 | 0.063 | 6.6% | 0.000 | 0.0% | 0.032 | 3.3% | 0.071 | 7.4%  |
|   |  Low Positive | 80 | 1.2 | 0.079 | 6.6% | 0.000 | 0.0% | 0.027 | 2.2% | 0.083 | 7.0%  |
|   |  Positive | 80 | 1.5 | 0.089 | 5.9% | 0.047 | 3.1% | 0.026 | 1.7% | 0.105 | 6.8%  |
|   |  Positive | 80 | 2.6 | 0.158 | 6.1% | 0.000 | 0.0% | 0.000 | 0.0% | 0.158 | 6.1%  |
|   |  Positive | 80 | 3.1 | 0.150 | 4.8% | 0.070 | 2.2% | 0.039 | 1.3% | 0.170 | 5.5%  |

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|  BioPlex 2200 CMV IgM |   |   |   | Within Run |   | Between-Run |   | Between-Day |   | Total  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  Sample Type | CMV IgM Panel Members | N | Mean (AI) | SD | %CV | SD | %CV | SD | %CV | SD | %CV  |
|  Serum | Negative | 80 | 0.4 | 0.039 | 10.3% | 0.019 | 5.2% | 0.005 | 1.5% | 0.044 | 11.6%  |
|   |  High Negative | 80 | 0.8 | 0.060 | 7.5% | 0.000 | 0.0% | 0.017 | 2.2% | 0.063 | 7.8%  |
|   |  Cut-Off | 80 | 1.1 | 0.065 | 6.1% | 0.030 | 2.8% | 0.032 | 3.0% | 0.078 | 7.4%  |
|   |  Cut-Off | 80 | 1.2 | 0.152 | 12.2% | 0.000 | 0.0% | 0.000 | 0.0% | 0.152 | 12.2%  |
|   |  Low Positive | 80 | 1.2 | 0.089 | 7.2% | 0.000 | 0.0% | 0.000 | 0.0% | 0.089 | 7.2%  |
|   |  Positive | 80 | 1.7 | 0.122 | 7.0% | 0.000 | 0.0% | 0.000 | 0.0% | 0.122 | 7.0%  |
|   |  Positive | 80 | 3.0 | 0.196 | 6.6% | 0.000 | 0.0% | 0.030 | 1.0% | 0.198 | 6.7%  |
|   |  Positive | 80 | 3.1 | 0.214 | 7.0% | 0.000 | 0.0% | 0.077 | 2.5% | 0.227 | 7.4%  |
|  Potassium EDTA | Negative | 80 | 0.3 | 0.047 | 13.8% | 0.000 | 0.0% | 0.022 | 6.5% | 0.052 | 15.3%  |
|   |  High Negative | 80 | 0.8 | 0.047 | 6.2% | 0.030 | 3.9% | 0.015 | 1.9% | 0.058 | 7.5%  |
|   |  Cut-Off | 80 | 1.1 | 0.070 | 6.5% | 0.000 | 0.0% | 0.028 | 2.6% | 0.075 | 7.0%  |
|   |  Low Positive | 80 | 1.3 | 0.089 | 6.7% | 0.000 | 0.0% | 0.000 | 0.0% | 0.089 | 6.7%  |
|   |  Positive | 80 | 1.9 | 0.128 | 6.8% | 0.000 | 0.0% | 0.078 | 4.1% | 0.150 | 8.0%  |
|   |  Positive | 80 | 3.1 | 0.185 | 6.0% | 0.000 | 0.0% | 0.088 | 2.9% | 0.205 | 6.7%  |
|   |  Positive | 80 | 2.9 | 0.196 | 6.7% | 0.000 | 0.0% | 0.122 | 4.2% | 0.231 | 7.9%  |
|  Sodium Heparin | Negative | 80 | 0.3 | 0.022 | 7.5% | 0.000 | 0.0% | 0.000 | 0.0% | 0.022 | 7.5%  |
|   |  High Negative | 80 | 0.8 | 0.065 | 7.8% | 0.000 | 0.0% | 0.000 | 0.0% | 0.065 | 7.8%  |
|   |  Cut-Off | 80 | 0.8 | 0.047 | 6.3% | 0.022 | 3.0% | 0.017 | 2.3% | 0.055 | 7.3%  |
|   |  Low Positive | 80 | 1.3 | 0.083 | 6.2% | 0.019 | 1.4% | 0.000 | 0.0% | 0.085 | 6.3%  |
|   |  Positive | 80 | 2.0 | 0.121 | 6.0% | 0.000 | 0.0% | 0.048 | 2.4% | 0.130 | 6.5%  |
|   |  Positive | 80 | 3.1 | 0.193 | 6.2% | 0.000 | 0.0% | 0.064 | 2.1% | 0.203 | 6.5%  |
|   |  Positive | 80 | 3.1 | 0.167 | 5.4% | 0.000 | 0.0% | 0.076 | 2.5% | 0.184 | 6.0%  |
|  Lithium Heparin | Negative | 80 | 0.3 | 0.040 | 14.2% | 0.000 | 0.0% | 0.000 | 0.0% | 0.040 | 14.2%  |
|   |  High Negative | 80 | 0.7 | 0.060 | 8.8% | 0.000 | 0.0% | 0.022 | 3.3% | 0.064 | 9.4%  |
|   |  Cut-Off | 80 | 1.0 | 0.085 | 8.7% | 0.000 | 0.0% | 0.037 | 3.7% | 0.093 | 9.5%  |
|   |  Low Positive | 80 | 1.4 | 0.104 | 7.5% | 0.000 | 0.0% | 0.055 | 4.0% | 0.118 | 8.5%  |
|   |  Positive | 80 | 1.9 | 0.133 | 7.1% | 0.019 | 1.0% | 0.042 | 2.2% | 0.140 | 7.6%  |
|   |  Positive | 80 | 3.0 | 0.222 | 7.5% | 0.096 | 3.2% | 0.000 | 0.0% | 0.242 | 8.1%  |
|   |  Positive | 80 | 2.7 | 0.153 | 5.7% | 0.030 | 1.1% | 0.000 | 0.0% | 0.156 | 5.8%  |

# Reproducibility:

A reproducibility panel, consisting of six (6) panel members made using serum matrix and BioPlex ToRC IgM QC controls was prepared by Bio-Rad Laboratories. The serum sample panel included one (1) low negative, one (1) high negative, one (1) low positive near cut-off, one (1) medium positive, and one (1) high positive sample, and one (1) positive control for all three (3) analytes. Reproducibility testing was performed at three (3) US testing facilities using one (1) lot of the BioPlex 2200 ToRC IgM Pack, one (1) lot of BioPlex 2200 ToRC IgM Calibrator Set and one (1) lot of BioPlex 2200 ToRC IgM Control Set. The panels were provided to each of the testing sites. Each of the panel members and control sets were tested in replicates of four (4) on two runs per day over five (5) days at three (3) sites (4 replicates x 2 runs

{9}

x 5 days x 3 Sites = 120 replicates per panel member). The data were analyzed for within-run (repeatability), between-run, between-day, between-site and total precision according to the principles described in Clinical Laboratory Standards Institute (CLSI) EP15-A3. The grand mean, standard deviation (SD) and percent coefficient of variation (% CV) were calculated.

## BioPlex 2200 Toxo IgM

|  T. gondii IgM
Panel Member | Mean
(AI) | Within Run |   | Between Run |   | Between Day |   | Between Site |   | Total  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |  SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV  |
|  Positive Control | 2.0 | 0.092 | 4.7% | 0.103 | 5.2% | 0.000 | 0.0% | 0.178 | 9.0% | 0.225 | 11.4%  |
|  Low Negative | 0.2 | 0.029 | 13.8% | 0.016 | 7.6% | 0.000 | 0.0% | 0.010 | 4.7% | 0.035 | 16.5%  |
|  High Negative | 0.8 | 0.040 | 4.8% | 0.012 | 1.5% | 0.028 | 3.3% | 0.100 | 12.0% | 0.112 | 13.4%  |
|  Low Positive | 1.3 | 0.051 | 4.0% | 0.032 | 2.5% | 0.023 | 1.8% | 0.148 | 11.6% | 0.162 | 12.7%  |
|  Mid Positive | 2.2 | 0.081 | 3.7% | 0.058 | 2.6% | 0.032 | 1.5% | 0.257 | 11.7% | 0.278 | 12.7%  |
|  High Positive | 3.2 | 0.172 | 5.3% | 0.093 | 2.9% | 0.043 | 1.3% | 0.280 | 8.7% | 0.344 | 10.7%  |

## BioPlex 2200 Rubella IgM

|  Rubella IgM
Panel Member | Mean
(AI) | Within Run |   | Between Run |   | Between Day |   | Between Site |   | Total  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |  SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV  |
|  Positive Control | 1.9 | 0.086 | 4.5% | 0.053 | 2.8% | 0.043 | 2.3% | 0.198 | 10.5% | 0.227 | 12.0%  |
|  Low Negative | 0.3 | 0.026 | 9.3% | 0.013 | 4.7% | 0.000 | 0.0% | 0.042 | 15.3% | 0.051 | 18.5%  |
|  High Negative | 0.8 | 0.048 | 6.2% | 0.000 | 0.0% | 0.026 | 3.3% | 0.097 | 12.4% | 0.111 | 14.2%  |
|  Low Positive | 1.2 | 0.056 | 4.8% | 0.019 | 1.7% | 0.000 | 0.0% | 0.143 | 12.3% | 0.155 | 13.3%  |
|  Mid Positive | 1.5 | 0.051 | 3.5% | 0.014 | 1.0% | 0.027 | 1.8% | 0.197 | 13.6% | 0.206 | 14.2%  |
|  High Positive | 2.8 | 0.139 | 4.9% | 0.015 | 0.5% | 0.030 | 1.1% | 0.328 | 11.6% | 0.358 | 12.6%  |

## BioPlex 2200 CMV IgM

|  CMV IgM
Panel Member | Mean
(AI) | Within Run |   | Between Run |   | Between Day |   | Between Site |   | Total  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |  SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV  |
|  Positive Control | 2.3 | 0.098 | 4.3% | 0.053 | 2.4% | 0.058 | 2.6% | 0.145 | 6.4% | 0.192 | 8.5%  |
|  Low Negative | 0.3 | 0.032 | 9.1% | 0.000 | 0.0% | 0.013 | 3.8% | 0.052 | 15.1% | 0.063 | 18.1%  |
|  High Negative | 0.7 | 0.048 | 7.0% | 0.011 | 1.6% | 0.009 | 1.3% | 0.062 | 9.0% | 0.080 | 11.6%  |
|  Low Positive | 1.2 | 0.071 | 6.2% | 0.029 | 2.5% | 0.000 | 0.0% | 0.096 | 8.3% | 0.123 | 10.6%  |
|  Mid Positive | 1.7 | 0.081 | 4.7% | 0.021 | 1.3% | 0.038 | 2.3% | 0.208 | 12.2% | 0.227 | 13.4%  |
|  High Positive | 2.9 | 0.146 | 5.1% | 0.026 | 0.9% | 0.065 | 2.3% | 0.257 | 9.0% | 0.304 | 10.6%  |

b. Linearity/assay reportable range:

Not Applicable

{10}

c. Traceability, Stability, Expected values (controls, calibrators, or methods):

# Calibrator Traceability

The BioPlex ToRC IgM calibrators are traceable against frozen internal standards which are anchored to a quantitation panel. The quantitation panel consists of patient samples whose analyte values span the assay range.

The BioPlex 2200 ToRC IgM Reagent Kit is calibrated using a set of two distinct serum based calibrators. A negative calibrator and a multianalyte  $T.$  gondii, Rubella CMV IgM calibrator are used to calibrate the  $T.$  gondii, Rubella and CMV IgM assays. The cut-off value and assignment of the calibrators were determined by determining the 98-99th percentile on normal samples and performing concordance and Receiver Operator Characteristic (ROC) analysis during feasibility and development.

The manufacturing target ranges of the Calibrator Set are listed below.

|  Calibrator Set | Range (AI)  |
| --- | --- |
|  Calibrator Level 1 | 0.0 – 0.2  |
|  Calibrator Level 2 | 1.1 – 2.0  |

# Controls

The control set includes two vials each of a negative control and a multianalyte positive control that contains  $T.$  gondii, Rubella and CMV IgM. The positive control contains known concentrations of human  $T.$  gondii, Rubella and CMV IgM, and is prepared by blending human disease state serum ( $T.$  gondii and CMV) and human recombinant Rubella IgM (huCal) with ToRC IgM negative serum matrix.

The assignment for the BioPlex 2200 ToRC IgM Control Set is performed using a minimum of two reagent lots along with matched calibrators with a target of three reagent lots with matched calibrators. For each control lot, three vials per control level are tested in replicates of five on all reagent lots for a total of fifteen replicates per reagent lot. This testing is performed on three analyzers yielding a total of forty-five replicates per reagent lot. The total number of replicates for each control level is 90 when two reagent lots are used and 135 when three reagent lots are used.

The manufacturing target ranges of the Control Set are listed below.

|  Control Set | Range (AI)  |
| --- | --- |
|  Negative Control | 0.0 – 0.5  |
|  Positive Control | 2.0 – 2.6  |

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12

## Kit Stability

BioPlex 2200 ToRC IgM Kit: Real Time (unopened) Kit Stability, 12 months or until the date of expiration when stored unopened on the instrument or at 2 to 8°C; the open kit claim is 60 days.

## Calibrator and Control Stability

BioPlex 2200 ToRC IgM Control and Calibrator Sets:

- Calibrator Open Vial Stability (2 to 8°C), 60 days from first opening;
- Control Open Vial Stability (2 to 8°C), 60 days from first opening;
- Onboard Calibration Curve Stability, 30 days;
- Real Time Stability of calibrators and controls (2 to 8°C), 12 months; labeled as until expiration date;
- Calibrators and Controls Accelerated Stability (2 to 8°C), 18 months predicted;
- Calibrators and Controls Freeze-thaw (-20°C or -70°C), 5 freeze thaw cycles.

## Sample Stability

Serum or plasma (K3 EDTA, lithium heparin, or sodium heparin) samples may be stored at room temperature (18 – 30°C) for up to 3 days and under refrigeration (2 – 8°C) for up to 7 days. For longer storage of samples, keep at -20°C or colder. Up to 5-freeze thaw cycles at -20°C and -70°C is acceptable.

## d. Detection limit:

Not Applicable

## e. Analytical specificity:

### Cross Reactivity:

The study was conducted to determine if samples from various disease states interfere with test results when tested with the BioPlex 2200 ToRC IgM kit. A panel of at least ten (10) specimens that are positive for each cross reactant were evaluated for possible cross reactivity with the BioPlex 2200 ToRC IgM kit for each of the three antibody assays. The potentially cross reactive samples were tested in commercially available predicate kits in order to confirm their negative status for the target analyte.

Cross reactivity, expressed as percent negative agreement is calculated by the ratio of the number of negative results to the total number of samples assayed for each cross reactant set of samples. The results of each potential cross reactant are listed below.

{12}

BioPlex 2200 ToRC IgM Cross Reactivity

|  Potential Cross Reactant | T. gondii IgM |   |   | Rubella IgM |   |   | CMV IgM  |   |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |  N | Neg | Pos | N | Neg | Pos | N | Neg | Pos  |
|  ANA Screen | 19 | 19 | 0 | 19 | 19 | 0 | 19 | 19 | 0  |
|  CMV IgM | 10 | 10 | 0 | 10 | 10 | 0 | N/A | N/A | 0  |
|  EBV IgM | 11 | 11 | 0 | 11 | 11 | 0 | 11 | 11 | 0  |
|  HAMA | 15 | 15 | 0 | 15 | 15 | 0 | 15 | 15 | 0  |
|  hCG | 12 | 12 | 0 | 12 | 12 | 0 | 12 | 12 | 0  |
|  HIV | 10 | 10 | 0 | 10 | 10 | 0 | 10 | 10 | 0  |
|  HSV-1/2 IgM | 13 | 13 | 0 | 13 | 13 | 0 | 13 | 13 | 0  |
|  Hypergamma-globulinemia IgM | 21 | 21 | 0 | 21 | 20 | 1 | 21 | 21 | 0  |
|  Influenza | 18 | 17 | 1 | 18 | 18 | 0 | 18 | 18 | 0  |
|  Measles IgM | 16 | 16 | 0 | 16 | 16 | 0 | 16 | 16 | 0  |
|  Mumps IgM | 13 | 12 | 1 | 13 | 13 | 0 | 13 | 13 | 0  |
|  Multiple Myeloma | 17 | 17 | 0 | 17 | 17 | 0 | 17 | 17 | 0  |
|  Parvovirus B 19 IgM | 14 | 14 | 0 | 14 | 13 | 1 | 14 | 14 | 0  |
|  Rheumatoid Factor | 11 | 11 | 0 | 11 | 11 | 0 | 11 | 11 | 0  |
|  Rubella IgM | 16 | 16 | 0 | N/A | N/A | 0 | 16 | 16 | 0  |
|  Toxo IgM | N/A | N/A | 0 | 10 | 10 | 0 | 10 | 10 | 0  |
|  VZV IgM | 13 | 12 | 1 | 13 | 13 | 0 | 13 | 12 | 1  |

Interfering Substances:

An interfering substances study was conducted to evaluate the potential interference of specific endogenous and exogenous substances with the BioPlex 2200 ToRC IgM assay according to the CLSI EP7-A2 guideline.

No significant 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  |

{13}

|  Substance | Concentration  |
| --- | --- |
|  Triglycerides | ≤ 3300 mg/dL  |
|  Beta Carotene | ≤ 0.6 mg/dL  |
|  Protein (total) | ≤ 12 g/dL  |
|  Ascorbic Acid | ≤ 6 mg/dL  |
|  Sodium Heparin | ≤ 8000 units/dL  |
|  Lithium Heparin | ≤ 8000 units/dL  |
|  EDTA (K2 and K3) | ≤ 800 mg/dL  |

f. Assay cut-off:

A final cut-off of 1.0 AI was established for the BioPlex 2200 ToRC IgM assay based on an evaluation of 401, 454 and 511 test-ordered and retrospective positive samples for  $T.$  gondii, Rubella and CMV IgM, respectively, for which the serological status was determined from the predicate  $T.$  gondii, Rubella and CMV IgM assays. The assay employs an equivocal zone that brackets the cut-off, which results in samples  $> 1.1$  AI being positive and  $< 0.9$  AI being negative. This analysis was used to optimize sensitivity and specificity for the BioPlex 2200 ToRC IgM assay.

2. Comparison studies:

a. Method comparison with predicate device:

Not Applicable

b. Matrix comparison:

Matched serum and plasma (EDTA and heparin sodium) samples drawn from the same donor were acquired from an outside reference lab. For each assay in the panel, a minimum of 40 sets of paired serum and plasma samples were prepared and values within the measurement range of the assay were analyzed in accordance with CLSI EP09-A3. Samples were assayed in replicates of two (2) with the second replicate run in reverse order. Mean plasma AI values were compared to matched mean serum AI values. Linear regression analysis was used to determine the presence of a matrix effect when compared to serum. The regression correlation parameters for slope, intercept and correlation coefficient (r) are shown below.

{14}

|  Matrix Comparison | N | BioPlex ToRC IgM Assay | Slope (95% CI) | Intercept (95% CI) | Correlation (r)  |
| --- | --- | --- | --- | --- | --- |
|  K3 EDTA vs. Serum | 54 | T. gondii IgM | 1.02 (0.99 to 1.06) | -0.05 (-0.12 to 0.02) | 0.994  |
|   |  53 | Rubella IgM | 1.03 (1.00 to 1.06) | -0.02 (-0.08 to 0.05) | 0.994  |
|   |  60 | CMV IgM | 1.01 (0.97 to 1.05) | -0.02 (-0.10 to 0.05) | 0.989  |
|  Lithium Heparin vs. Serum | 54 | T. gondii IgM | 1.02 (0.98 to 1.06) | -0.04 (-0.12 to 0.04) | 0.992  |
|   |  51 | Rubella IgM | 1.03 (1.00 to 1.06) | -0.02 (-0.08 to 0.04) | 0.995  |
|   |  60 | CMV IgM | 0.98 (0.93 to 1.02) | 0.00 (-0.09 to 0.09) | 0.985  |
|  Sodium Heparin vs. Serum | 54 | T. gondii IgM | 1.03 (0.99 to 1.06) | -0.05 (-0.12 to 0.02) | 0.994  |
|   |  51 | Rubella IgM | 1.01 (0.97 to 1.05) | -0.01 (-0.09 to 0.06) | 0.993  |
|   |  59 | CMV IgM | 0.99 (0.96 to 1.03) | -0.04 (-0.11 to 0.04) | 0.991  |

# 3. Clinical studies:

a. Clinical Sensitivity:

Not Applicable

b. Clinical specificity:

Not Applicable

c. Other clinical supportive data (when a. and b. are not applicable):

# Prospective Study:

The performance of the BioPlex 2200 ToRC IgM kit was tested against corresponding commercially available predicate  $T$  gondii, Rubella and CMV IgM assays. A total of 2,129 prospective samples (approximately 700 samples per analyte) submitted for  $T$  gondii, Rubella, or CMV testing were tested at 3 U.S. clinical testing sites. Of the approximate 700 samples per analyte, 200 were from pregnant women for each analyte.

Results from all sites are shown and summarized in the table below.

{15}

BioPlex 2200 ToRC IgM vs Commercially Available Immunoassay (Prospective Study)

|  Test Ordered | BioPlex 2200 ToRC IgM Assay  |   |   |   |   |   |   |   |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |   |   |  Pos (+) | Eqv | Neg (-) | Total | Pos (+) % Agreement 95% CI | Neg (-) % Agreement 95% CI  |
|  Commercially Available Immunoassay | T. gondii IgM | Pregnant Women | Pos (+) | 0 | 0 | 0 | 0 | N/A | 98.0% (196/200) 95.0 to 99.2%  |
|   |   |   |  Eqv | 0 | 0 | 0 | 0  |   |   |
|   |   |   |  Neg (-) | 3 | 1 | 196 | 200  |   |   |
|   |   |   |  Total | 3 | 1 | 196 | 200  |   |   |
|   |   |  Test Ordered | Pos (+) | 0 | 0 | 0 | 0 | N/A | 97.4% (481/494) 95.6 to 98.5%  |
|   |   |   |  Eqv | 1 | 0 | 0 | 1  |   |   |
|   |   |   |  Neg (-) | 9 | 3 | 481 | 493  |   |   |
|   |   |   |  Total | 10 | 3 | 481 | 494  |   |   |
|   |  Rubella IgM | Pregnant Women | Pos (+) | 0 | 0 | 0 | 0 | N/A | 100.0% (198/198) 98.1 to 100.0%  |
|   |   |   |  Eqv | 0 | 0 | 2 | 2  |   |   |
|   |   |   |  Neg (-) | 0 | 0 | 198 | 198  |   |   |
|   |   |   |  Total | 0 | 0 | 200 | 200  |   |   |
|  Test Ordered |   | Pos (+) | 4 | 1 | 2 | 7 | 40.0% (4/10)* 16.8 to 68.7% | 99.6% (498/500) 98.6 to 99.9%  |   |
|   |   |  Eqv | 0 | 2 | 3 | 5  |   |   |   |
|   |   |  Neg (-) | 1 | 1 | 498 | 500  |   |   |   |
|   |   |  Total | 5 | 4 | 503 | 512  |   |   |   |
|  CMV IgM | Pregnant Women | Pregnant Women | Pos (+) | 8 | 2 | 4 | 14 | 50.0% (8/16)** 28.0 to 72.0% | 100.0% (183/183) 97.9 to 100.0%  |
|   |   |   |  Eqv | 0 | 1 | 2 | 3  |   |   |
|   |   |   |  Neg (-) | 0 | 0 | 183 | 183  |   |   |
|   |   |   |  Total | 8 | 3 | 189 | 200  |   |   |
|   |   |  Test Ordered | Pos (+) | 20 | 1 | 11 | 32 | 55.6% (20/36)*** 39.6 to 70.5% | 98.6% (480/487) 97.1 to 99.3%  |
|   |   |   |  Eqv | 0 | 0 | 4 | 4  |   |   |
|   |   |   |  Neg (-) | 3 | 4 | 480 | 487  |   |   |
|   |   |   |  Total | 23 | 5 | 495 | 523  |   |   |

* Three samples were predicate equivocal and BioPlex Rubella IgM negative, two samples were predicate positive and BioPlex Rubella IgM negative and one sample was predicate positive and BioPlex Rubella IgM equivocal.
** Of the sixteen samples, Six samples were negative and three were equivocal by BioPlex CMV IgM. Eight samples were negative and one equivocal by another FDA cleared device.

{16}

*** Eleven positive and four equivocal results for the predicate were all negative by BioPlex CMV IgM. One positive result for the predicate was equivocal by BioPlex CMV IgM. All but three were confirmed negative by another FDA cleared device.

## Retrospective Study:

Performance of the BioPlex 2200 ToRC IgM kit was also evaluated against corresponding commercially available *T. gondii*, Rubella, and CMV IgM immunoassays using presumptive positive samples. Three clinical sites tested 210 *T. gondii* (134 female, 76 male), 101 Rubella (44 female, 57 male) and 213 CMV (119 female, 94 male) IgM presumptive positive samples. Presumed positive banked samples for ToRC IgM were further selected by the respective predicate device use for the comparative analysis. The characteristics of samples with presumptive positive status are shown below.

### BioPlex 2200 ToRC IgM vs Commercially Available Immunoassay (Retrospective Study)

|  Presumptive Positive for *T. gondii*, Rubella or CMV IgM | BioPlex 2200 ToRC IgM Agreement  |   |   |   |   |   |   |
| --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  Pos (+) | Eqv | Neg (-) | Total | Pos (+) % Agreement 95% CI  |
|  Commercially Available Immunoassay | T. gondii IgM | Pos (+) | 203 | 3 | 3 | 209 | 97.1% (203/209)
93.9 to 98.7%  |
|   |   |  Eqv | 1 | 0 | 0 | 1  |   |
|   |   |  Neg (-) | 0 | 0 | 0 | 0  |   |
|   |   |  Total | 204 | 3 | 3 | 210  |   |
|   |  Rubella IgM | Pos (+) | 96 | 1 | 1 | 98 | 98.0% (96/98)
92.9 to 99.4%  |
|   |   |  Eqv | 1 | 0 | 0 | 1  |   |
|   |   |  Neg (-) | 1 | 0 | 1 | 2  |   |
|   |   |  Total | 98 | 1 | 2 | 101  |   |
|   |  CMV IgM | Pos (+) | 198 | 2 | 1 | 201 | 98.5% (198/201)
95.7 to 99.5%  |
|   |   |  Eqv | 4 | 1 | 0 | 5  |   |
|   |   |  Neg (-) | 4 | 0 | 3 | 7  |   |
|   |   |  Total | 206 | 3 | 4 | 213  |   |

## Correlation with CDC Evaluation Panels:

A correlation study was performed to evaluate the characteristics of the BioPlex 2200 ToRC IgM kit with serum panels provided by the Centers for Disease Control and Prevention (CDC) for *T. gondii* IgM. The results are presented as a means to convey further information on the performance. This does not imply an endorsement of the BioPlex 2200 ToRC IgM kit by the CDC. Results are presented in the table below.

{17}

Results From Testing of CDC T. gondii Reference Sera (N=97)

|  CDC Panel
T. gondii
IgM | BioPlex T. gondii IgM |   |   |   |  |   |
| --- | --- | --- | --- | --- | --- | --- |
|   |  Pos(+) | Eq | Neg(-) | Total | Positive Agreement | Negative Agreement  |
|  Pos (+) | 32 | 0 | 0 | 32 | 100.0%
89.3 to 100.0 | 100.0%
94.4 to 100.0  |
|  Neg (-) | 0 | 0 | 65 | 65  |   |   |
|  Total | 32 | 0 | 65 | 97  |   |   |

Seroconversion Testing:

Commercially available seroconversion panels for T. gondii, Rubella and CMV IgM were tested with the BioPlex 2200 ToRC IgM kit and compared with the predicate commercial method.

BioPlex 2200 ToRC IgM Kit, Comparison to Commercial Method - T. gondii IgM

|  Panel | Day | T. gondii IgM  |   |
| --- | --- | --- | --- |
|   |   |  VIDAS Toxo IgM | BioPlex ToRC IgM  |
|   |   |  Index | Antibody Index (AI)  |
|  Antibody Systems | 0 | 0.24(Neg) | <0.2(Neg)  |
|   |  7 | 1.62(Pos) | 2.1(Pos)  |
|   |  9 | 3.24(Pos) | 3.3(Pos)  |
|   |  14 | 5.78(Pos) | >4.0(Pos)  |
|   |  16 | 5.80(Pos) | >4.0(Pos)  |
|   |  23 | 5.38(Pos) | >4.0(Pos)  |
|   |  27 | 5.24(Pos) | >4.0(Pos)  |
|   |  30 | 5.04(Pos) | >4.0(Pos)  |
|   |  49 | 4.59(Pos) | >4.0(Pos)  |
|   |  53 | 4.35(Pos) | >4.0(Pos)  |
|   |  86 | 3.30(Pos) | >4.0(Pos)  |
|   |  88 | 2.94(Pos) | 3.6(Pos)  |
|   |  93 | 2.67(Pos) | 3.1(Pos)  |
|   |  95 | 2.62(Pos) | 2.7(Pos)  |
|   |  100 | 2.58(Pos) | 2.8(Pos)  |
|   |  104 | 2.34(Pos) | 2.6(Pos)  |
|   |  107 | 2.36(Pos) | 2.2(Pos)  |
|   |  112 | 2.28(Pos) | 2.4(Pos)  |
|   |  114 | 2.21(Pos) | 2.3(Pos)  |
|   |  119 | 2.29(Pos) | 2.1(Pos)  |

{18}

BioPlex 2200 ToRC IgM Kit, Comparison to Commercial Method – Rubella IgM

|  Panel | Day | Rubella IgM  |   |
| --- | --- | --- | --- |
|   |   |  BioPlex RC IgM | BioPlex ToRC IgM  |
|   |   |  Antibody Index (AI) | Antibody Index (AI)  |
|  Liquicheck - RP011 | 0 | <0.2 (Neg) | <0.2 (Neg)  |
|   |  3 | <0.2 (Neg) | <0.2 (Neg)  |
|   |  9 | <0.2 (Neg) | <0.2 (Neg)  |
|   |  12 | <0.2 (Neg) | <0.2 (Neg)  |
|   |  16 | 1.3 (Pos) | 1.0 (Eq)  |
|   |  19 | >4.0 (Pos) | >4.0 (Pos)  |
|   |  24 | >4.0 (Pos) | >4.0 (Pos)  |
|   |  27 | >4.0 (Pos) | >4.0 (Pos)  |
|   |  31 | 3.6 (Pos) | 3.4 (Pos)  |
|   |  36 | 2.1 (Pos) | 2.0 (Pos)  |
|   |  39 | 1.5 (Pos) | 1.4 (Pos)  |
|   |  43 | 1.0 (Eq) | 1.0 (Eq)  |
|   |  46 | 0.7 (Neg) | 0.6 (Neg)  |
|   |  50 | 0.6 (Neg) | 0.5 (Neg)  |
|   |  53 | 0.3 (Neg) | 0.4 (Neg)  |
|   |  57 | 0.3 (Neg) | 0.4 (Neg)  |
|   |  60 | 0.3 (Neg) | 0.3 (Neg)  |
|   |  64 | 0.2 (Neg) | 0.2 (Neg)  |
|   |  67 | 0.2 (Neg) | 0.2 (Neg)  |
|   |  71 | 0.2 (Neg) | 0.2 (Neg)  |

{19}

BioPlex 2200 ToRC IgM Kit, Comparison to Commercial Method - CMV IgM

|  Panel | Day | CMV IgM  |   |
| --- | --- | --- | --- |
|   |   |  BioPlex RC IgM | BioPlex ToRC IgM  |
|   |   |  Antibody Index (AI) | Antibody Index (AI)  |
|  Liquicheck - RP003 | 1 | >4.0 (Pos) | 2.8 (Pos)  |
|   |  4 | >4.0 (Pos) | >4.0 (Pos)  |
|   |  8 | >4.0 (Pos) | >4.0 (Pos)  |
|   |  51 | 1.9 (Pos) | 2.3 (Pos)  |
|   |  55 | 1.8 (Pos) | 2.1 (Pos)  |
|   |  59 | 1.2 (Pos) | 1.8 (Pos)  |
|   |  65 | 1.3 (Pos) | 1.5 (Pos)  |
|   |  67 | 1.4 (Pos) | 1.5 (Pos)  |
|   |  72 | 1.3 (Pos) | 1.1 (Pos)  |
|   |  74 | 1.1 (Pos) | 1.0 (Eq)  |
|   |  79 | 1.6 (Pos) | 1.2 (Pos)  |
|   |  84 | 1.4 (Pos) | 1.3 (Pos)  |
|   |  88 | 1.3 (Pos) | 1.4 (Pos)  |
|   |  95 | 1.3 (Pos) | 1.1 (Pos)  |
|   |  99 | 1.4 (Pos) | 1.0 (Eq)  |

IgM Specificity:

Samples positive for  $T.$  gondii, Rubella and CMV IgM and IgG were treated with dithiothreitol (DTT) to inactivate IgM activity. The samples were assayed neat and diluted into assay range in replicates of two. IgM was measured using the BioPlex 2200 ToRC IgM kit.

BioPlex ToRC IgM - T. gondii IgM Specificity

|  Sample ID | T. gondii IgM (AI) Before DTT | DTT Treatment AI (% recovery)  |
| --- | --- | --- |
|  Sample 1 | 9.5 | 0.1 (1.05%)  |
|  Sample 2 | 9.5 | 0.1 (1.05%)  |
|  Sample 3 | 18.0 | 0.3 (1.67%)  |
|  Sample 4 | 15.5 | 0.5 (3.23%)  |
|  Sample 5 | 9.0 | 0.1 (1.11%)  |
|  Sample 6 | 14.0 | 0.2 (1.43%)  |
|  Sample 7 | 14.0 | 0.1 (0.71%)  |
|  Sample 8 | 15.0 | 0.1 (0.67%)  |
|  Sample 9 | 15.0 | 0.1 (0.67%)  |
|  Sample 10 | 16.5 | 0.1 (0.61%)  |

{20}

BioPlex ToRC IgM – Rubella IgM Specificity

|  Sample ID | Rubella IgM (AI) Before DTT | DTT Treatment AI (% recovery)  |
| --- | --- | --- |
|  Sample 1 | 8.0 | 0.1 (1.25%)  |
|  Sample 2 | 9.0 | 0.0 (0.00%)  |
|  Sample 3 | 8.5 | 0.0 (0.00%)  |
|  Sample 4 | 5.0 | 0.1 (2.00%)  |
|  Sample 5 | 9.0 | 0.1 (1.11%)  |
|  Sample 6 | 7.0 | 0.1 (1.43%)  |
|  Sample 7 | 4.0 | 0.0 (0.00%)  |
|  Sample 8 | 5.0 | 0.0 (0.00%)  |
|  Sample 9 | 9.0 | 0.2 (2.22%)  |
|  Sample 10 | 6.0 | 0.1 (1.67%)  |

BioPlex ToRC IgM – CMV IgM Specificity

|  Sample ID | CMV IgM (AI) Before DTT | DTT Treatment AI (% recovery)  |
| --- | --- | --- |
|  Sample 1 | 25.0 | 0.1 (0.40%)  |
|  Sample 2 | 11.0 | 0.2 (1.82%)  |
|  Sample 3 | 8.5 | 0.1 (1.18%)  |
|  Sample 4 | 20.0 | 0.5 (2.50%)  |
|  Sample 5 | 15.5 | 0.2 (1.29%)  |
|  Sample 6 | 18.5 | 0.2 (1.08%)  |
|  Sample 7 | 21.0 | 0.2 (0.95%)  |
|  Sample 8 | 6.0 | 0.0 (0.00%)  |
|  Sample 9 | 16.5 | 1.2 (7.27%)  |
|  Sample 10 | 31.5 | 0.7 (2.22%)  |

4. Clinical cut-off:

Not Applicable

5. Expected values/Reference range:

Expected values for the BioPlex 2200 ToRC IgM kit are presented by age and gender in the following tables for samples from pregnant women and patients sent to the lab for *T. gondii*, Rubella or CMV IgM testing. A total of two hundred (200) each of pregnant women samples sent to the lab for *T. gondii*, Rubella or CMV IgM testing and approximately five hundred (500) each of samples sent to the lab for *T. gondii*, Rubella or CMV IgM testing were tested.

{21}

BioPlex 2200 ToRC IgM Prevalence Results by Age and Gender: Samples from Pregnant Women Who Were Sent for T.gondii, Rubella, or CMV IgM Testing by Age Group

|  Age | T. gondii IgM |   | Rubella IgM |   | CMV IgM  |   |
| --- | --- | --- | --- | --- | --- | --- |
|   |  Pos/Total | % Prevalence | Pos/Total | % Prevalence | Pos/Total | % Prevalence  |
|  15-25 | 1/81 | 1.2% | 0/86 | 0.0% | 4/85 | 4.7%  |
|  26-35 | 2/98 | 2.0% | 0/93 | 0.0% | 4/100 | 4.0%  |
|  36-43 | 0/21 | 0.0% | 0/21 | 0.0% | 0/15 | 0.0%  |
|  Total | 3/200 | 1.5% | 0/200 | 0.0% | 8/200 | 4.0%  |

BioPlex 2200 ToRC IgM Prevalence Results by Age and Gender: Samples from Patients Who Were Sent for T.gondii, Rubella, or CMV IgM Testing

|  Age | Gender | T. gondii IgM |   | Rubella IgM |   | CMV IgM  |   |
| --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |  Pos/Total | % Prevalence | Pos/Total | % Prevalence | Pos/Total | % Prevalence  |
|  <1-90 | F | 5/357 | 1.4% | 3/357 | 0.8% | 14/325 | 4.3%  |
|   |  M | 5/137 | 3.6% | 2/155 | 1.3% | 9/198 | 4.5%  |
|  Total |   | 10/494 | 2.0% | 5/512 | 1.0% | 23/523 | 4.4%  |

N. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

---

**Source:** [https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/PUQ/K170509](https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/PUQ/K170509)

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