The BioPlex 2200 ToRC IgM kit is a multiplex flow immunoassay intended for the qualitative detection of IgM antibodies to Toxoplasma 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 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 Description

BioPlex ToRC IgM Reagent Pack includes the following components:

One (1) 10 mL vial, containing dyed beads coated with lysates of T. gondii, Rubella 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 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 for rubella and human disease state plasma derived antibodies for 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 set contains two (2) 1.5 mL Positive Control serum vials, containing human disease state plasma derived IgM antibodies to T. gondii and CMV, and HuCAL® recombinant IgM antibodies to Rubella 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 defibrinted 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%).

Additional materials required but not supplied include BioPlex 2200 Sheath Fluid containing Phosphate Buffered Saline (PBS), ProClin 300 (0.03%) and sodium azide (<0.1%) as preservatives; and BioPlex 2200 Wash Solution containing Phosphate Buffered Saline (PBS) and Tween 20, ProClin 300 (0.03%) and sodium azide (<0.1%) as preservatives.

AI/ML Overview

Here's an analysis of the provided text to extract information about the acceptance criteria and the study proving the device's performance, as requested.

The provided text describes the performance characteristics of the BioPlex 2200 ToRC IgM kit, which is a multiplex flow immunoassay for detecting IgM antibodies to Toxoplasma gondii, Rubella, and Cytomegalovirus (CMV).

Acceptance Criteria and Reported Device Performance

The acceptance criteria are not explicitly stated as a section titled "Acceptance Criteria" with pass/fail metrics. Instead, the document presents various analytical and clinical performance studies, and the results of these studies implicitly represent the device's acceptable performance. For the purpose of this response, I will infer the acceptance criteria from the reported performance, particularly where quantitative results are presented.

1. Table of Acceptance Criteria and Reported Device Performance

Performance Metric Category	Specific Metric (Inferred Acceptance Criteria)	Reported Device Performance	Comments
Analytical Performance	Precision/Reproducibility
	Within-run Precision	See tables below	Measured as SD for AI < 0.8 and %CV for AI > 0.8. Generally low %CVs (e.g., 3.7% to 14.2% for Negative samples, 3.8% to 7.2% for Positives).
	Between-run Precision	See tables below	Generally low %CVs across analytes and sample types.
	Between-day Precision	See tables below	Generally low %CVs across analytes and sample types.
	Total Reproducibility (Across Sites)	See tables below	Generally low %CVs (e.g., Toxo IgM Total %CV for high positive is 10.7%).
Analytical Specificity	Cross-Reactivity (Percent Negative Agreement)		Assessed by testing against various potential cross-reactants. Primarily 100% negative agreement, with few exceptions (e.g., Rubella IgM with Hypergamma-globulinemia IgM: 20/21 negative; Rubella IgM with Parvovirus B19 IgM: 13/14 negative; VZV IgM with T. gondii IgM and CMV IgM: 12/13 negative for both).
	Interfering Substances	No significant interference observed.	Tested substances include Hemoglobin, Bilirubin, Cholesterol, Red Blood Cells, Gamma Globulin, Triglycerides, Beta Carotene, Protein, Ascorbic Acid, Sodium Heparin, Lithium Heparin, EDTA.
Clinical Performance	Method Comparison (Prospective Samples)		Comparison against commercially available predicate devices.
	T. gondii IgM (Pregnant Women)	Positive Agreement: N/A, Negative Agreement: 98.0% (196/200) CI 95.0-99.2%	High negative agreement.
	T. gondii IgM (Test Ordered)	Positive Agreement: N/A, Negative Agreement: 97.4% (481/494) CI 95.6-98.5%	High negative agreement.
	Rubella IgM (Pregnant Women)	Positive Agreement: N/A, Negative Agreement: 100.0% (198/198) CI 98.1-100.0%	Excellent negative agreement.
	Rubella IgM (Test Ordered)	Positive Agreement: 40.0% (4/10) CI 16.8-68.7%, Negative Agreement: 99.6% (498/500) CI 98.6-99.9%	Lower positive agreement for test-ordered samples, but very high negative agreement. Note: 10 samples considered positive by predicate, 5 negative and 1 equivocal by BioPlex.
	CMV IgM (Pregnant Women)	Positive Agreement: 50.0% (8/16) CI 28.0-72.0%, Negative Agreement: 100.0% (183/183) CI 97.9-100.0%	Lower positive agreement for pregnant women, but excellent negative agreement. Discrepant samples were confirmed negative by another FDA-cleared device.
	CMV IgM (Test Ordered)	Positive Agreement: 55.6% (20/36) CI 39.6-70.5%, Negative Agreement: 98.6% (480/487) CI 97.1-99.3%	Lower positive agreement for test-ordered samples, but high negative agreement. Discrepant samples were confirmed negative by another FDA-cleared device.
	Method Comparison (Retrospective Samples - Presumptive Positive)		Comparison against commercially available predicate devices.
	T. gondii IgM	Positive Agreement: 97.1% (203/209) CI 93.9-98.7%	High positive agreement.
	Rubella IgM	Positive Agreement: 98.0% (96/98) CI 92.9-99.4%	High positive agreement.
	CMV IgM	Positive Agreement: 98.5% (198/201) CI 95.7-99.5%	High positive agreement.
Clinical Supportive Data	*Correlation with CDC Evaluation Panels (T. gondii* IgM)**	Positive Agreement: 100.0%, Negative Agreement: 100.0%	Excellent agreement with CDC reference sera.
	Seroconversion Testing	Qualitative agreement with predicate devices for seroconversion panels.	Demonstrates expected seroconversion patterns.
	IgM Specificity (DTT Treatment)	High reduction in IgM activity (very low % recovery - typically <3%).	Confirms specificity for IgM antibodies.

Precision Data Tables (as presented in the document):

BioPlex 2200 Toxo IgM – CLSI EP5-A3 Precision

Sample Type	T. gondii IgM Panel Members	N	Mean (AI)	SD	%CV	SD	%CV	SD	%CV	SD	%CV
				Within Run		Between Run		Between Day		Total
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%

BioPlex 2200 Rubella IgM – CLSI EP5-A3 Precision

Sample Type	Rubella IgM Panel Members	N	Mean (AI)	SD	%CV	SD	%CV	SD	%CV	SD	%CV
				Within Run		Between Run		Between-Day		Total
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%

BioPlex 2200 CMV IgM – CLSI EP5-A3 Precision

Sample Type	CMV IgM Panel Members	N	Mean (AI)	SD	%CV	SD	%CV	SD	%CV	SD	%CV
				Within Run		Between Run		Between-Day		Total
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 Data Tables (across 3 sites):

BioPlex 2200 Toxo IgM - CLSI EP15-A3 Reproducibility

Panel Member	Mean (AI)	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
		Within Run		Between Run		Between Day		Between Site		Total
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 - CLSI EP15-A3 Reproducibility

Panel Member	Mean (AI)	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
		Within Run		Between Run		Between Day		Between Site		Total
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 - CLSI EP15-A3 Reproducibility

CMV IgM Panel Member	Mean (AI)	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
		Within Run		Between Run		Between Day		Between Site		Total
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%

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

Precision/Reproducibility:
- CLSI EP5-A3 Precision: 80 data points per panel member for each analyte (T. gondii, Rubella, CMV) and each sample matrix (Serum, Potassium EDTA, Sodium Heparin, Lithium Heparin). This means 80 samples per specific condition.
- CLSI EP15-A3 Reproducibility: 120 replicates per panel member (4 replicates x 2 runs x 5 days x 3 sites). There were 6 panel members for each analyte. This equates to 720 samples per analyte across three sites for reproducibility.
Cross-Reactivity: At least 10 specimens per potential cross-reactant for each of the three antibody assays.
Matrix Comparison: A minimum of 40 sets of paired serum and plasma samples per analyte (N=51 to 60 for each analyte and matrix comparison).
Method Comparison (Prospective):
- 2129 prospective samples in total.
- ~700 samples per analyte (T. gondii, Rubella, CMV).
- 200 pregnant women samples for each analyte.
- Data Provenance: Samples were submitted for T. gondii, Rubella, or CMV testing. These were prospective samples collected at 3 U.S. clinical testing sites.
Method Comparison (Retrospective):
- 210 T. gondii IgM presumptive positive samples (134 female, 76 male).
- 101 Rubella IgM presumptive positive samples (44 female, 57 male).
- 213 CMV IgM presumptive positive samples (119 female, 94 male).
- Data Provenance: Presumed positive banked samples for ToRC IgM. Retrospective.
Correlation with CDC Evaluation Panels: 97 samples for T. gondii IgM (CDC Panel). Data provenance from CDC.
Seroconversion Testing: Commercially available seroconversion panels for T. gondii, Rubella, and CMV IgM.
IgM Specificity: 10 samples for each analyte (T. gondii, Rubella, CMV).

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

The document does not explicitly mention "experts" in the context of establishing ground truth for the test set. Instead, for clinical performance, the ground truth for comparative studies was established by:

Results from commercially available predicate T. gondii, Rubella, and CMV IgM assays.
In cases of discrepancy, another FDA-cleared device was used for confirmation (e.g., for CMV IgM method comparison).
For the CDC panel, the CDC's characterization of the samples served as the ground truth.

This implies that the "ground truth" relies on the established performance and regulatory clearance of existing in vitro diagnostic devices and reference panels, rather than expert human interpretation in the way it might for imaging studies. Healthcare professionals running these predicate assays are implied to be qualified clinical laboratory personnel, but no specific number or qualification of individual experts is provided as is typical for AI/imaging ground truth studies.

4. Adjudication Method for the Test Set

The document does not describe an adjudication method in the context of human readers reviewing results. For the comparative studies, discrepancies between the new device and the predicate device were investigated using another FDA-cleared device to confirm serological status. This serves as a form of adjudication by a third, independent, and validated method.

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. This is an in vitro diagnostic (IVD) device, not an AI for human-in-the-loop performance improvement study. The performance is assessed based on direct assay results (qualitative detection of antibodies), not on human reader performance. Therefore, an MRMC study or calculation of human reader improvement with AI assistance is not applicable and was not performed.

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

Yes. The entire performance evaluation for the BioPlex 2200 ToRC IgM kit is effectively a "standalone" assessment of the device's ability to qualitatively detect IgM antibodies. The device generates an Antibody Index (AI) result, which is then interpreted against a defined cut-off (e.g., >1.1 AI for positive, <0.9 AI for negative, 0.9-1.1 AI for equivocal). This is an algorithm-only performance in the context of an IVD, as it directly measures analytes in a sample.

7. The Type of Ground Truth Used

The primary ground truth used for method comparison studies was:

Results from established, commercially available, and presumably FDA-cleared predicate IVD devices.
For specific discrepant results, another FDA-cleared device was used, implying a reliance on a higher standard or independent confirmation through a validated method.
For T. gondii IgM, CDC reference sera panels were used, representing a type of reference standard/outcome data (serological status defined by reference lab/panel).
Seroconversion panels represent samples with known disease progression/antibody development over time, establishing dynamic ground truth.
DTT treatment for IgM specificity provides experimental evidence that the signal is specifically from IgM antibodies, using a chemical method to selectively inactivate IgM.

8. The Sample Size for the Training Set

The document does not explicitly describe a "training set" in the context of machine learning or AI. This is a traditional IVD device, and its performance is established through calibration and validation studies, not by training a machine learning model on a dataset.

However, the "Assay cut-off" section mentions: "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 samples for T. gondii, Rubella and CMV IgM, respectively, of which the serological status was determined from the predicate T. gondii, Rubella and CMV IgM assays."

This set of samples, used for cut-off optimization, somewhat resembles a "development set" or "tuning set" in machine learning, though it's integral to the assay's design and calibration rather than a separate training phase.

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

As noted in point 8, there isn't a "training set" in the AI/ML sense. For the samples used to establish the assay cut-off, the serological status (ground truth) was determined from the predicate T. gondii, Rubella, and CMV IgM assays. This indicates that commercially available and validated assays were used to define the true positive/negative status of these samples for optimizing the new device's cut-off.

Summary

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Public Health Service

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

July 25, 2017

BIO-RAD LABORATORIES ARLENE CARILLO REGULATORY AFFAIRS REPRESENTATIVE 5500 EAST SECOND STREET BENICIA, CA 94510

Re: K170509

Trade/Device Name: Bioplex 2200 ToRC IgM, Bioplex 2200 ToRC IgM Calibrator Set. Bioplex 2200 ToRC IgM Control Set Regulation Number: 21 CFR 866.3510 Regulation Name: Rubella virus serological reagents Regulatory Class: II Product Code: PUQ, JIX, JJX, LKQ, LFZ, LGD Dated: February 17, 2017 Received: February 21, 2017

Dear Ms. Carillo:

This letter corrects our substantially equivalent letter of May 19, 2017.

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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

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

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's 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 medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the

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electronic product radiation control provisions (Sections 531-542 of the Act): 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. 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 Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Steven R. Gitterman -S for

Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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510(k) Number (if known) K170509

Device Name BioPlex 2200 ToRC IgM Kit BioPlex 2200 ToRC IgM Calibrator Set BioPlex 2200 ToRC IgM Control Set

Indications for Use (Describe)

BioPlex 2200 ToRC IgM Kit

The BioPlex 2200 ToRC 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 ToRC IgM Calibrator Set

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

BioPlex 2200 ToRC IgM Control Set

Type of Use (Select one or both, as applicable)

|X Prescription Use (Part 21 CFR 801 Subpart D)

| | Over-The-Counter Use (21 CFR 801 Subpart C)

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BioPlex 2200 ToRC IgM 510(k) Summary

Bio-Rad Laboratories hereby submits this 510(k) in accordance with the requirements of SMDA 1990 and 21 CFR 807.92. This summary provides the information to support a determination of substantial equivalence for the BioPlex 2200 ToRC IgM kit.

510(k) Number: K170509

Summary Preparation Date: February 17, 2017

Applicant: Bio-Rad Laboratories

Contact: Arlene Carillo Regulatory Affairs Representative 5500 East Second Street Benicia, CA 94510

Purpose for Submission: New Device

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

Type of Test: Multiplexed Microparticle Immunoassay based on Luminex Technology

Proprietary and Established Names:

BioPlex 2200 ToRC IgM BioPlex 2200 ToRC IgM Calibrator Set BioPlex 2200 ToRC IgM Control Set

Product code	Classification	Regulation section	Panel
PUQ: Multiplexflow immunoassay,T. gondii , Rubella,CMV IgM	Class II	21 CFR § 866.3510 – Rubella virusSerological Agents	Microbiology
JIT, Calibrator,Secondary	Class II	21 CFR §862.1150 –Calibrator	Clinical Chemistry
JJY, Multi-AnalyteControls, All Kinds(Assayed)	Class I	21 CFR §862.1660 –Quality ControlMaterial (assayed	Clinical Chemistry

Regulatory Information:

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Product code	Classification	Regulation section	Panel
		and unassayed

The following is a list of regulation sections and product codes that are applicable to the individual analytes detected by the device subject of this submission.

21 CFR § 866.3780 – Toxoplasma gondii Serological Agents (Microbiology Panel: Class II), Product Code LGD, Enzyme Linked Immunoabsorbent Assay, Toxoplasma gondii 2. 21 CFR § 866.3510 - Rubella virus Serological Agents (Microbiology Panel: Class II), Product Code LFX, Enzyme Linked Immunoabsorbent Assay, Rubella 3. 21 CFR § 866.3175- Cytomegalovirus Serological Agents (Microbiology Panel: Class II), Product Code LFZ, Enzyme Linked Immunoabsorbent Assay, Cytomegalovirus and LKQ, Antibody IgM, if, Cytomegalovirus Virus

Intended Use:

1. Intended use(s):
  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 assay is not FDA cleared or approved for use in testing (screening) blood or plasma donors.

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

1. Indication(s) for use: Same as Intended Use
1. Special conditions for use statement(s): For prescription use only
1. Special instrument requirements: Bio-Rad BioPlex 2200 System

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Device Description:

BioPlex ToRC IgM Reagent Pack includes the following components:

One (1) 10 mL vial, containing dyed beads coated with lysates of T. gondii, Rubella 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.

Substantial Equivalence Information:

1. Predicate device name(s): BioPlex 2200 Rubella and CMV IgM Kit, K092587 bioMeriéux, Inc. VIDAS® TOXO IgM, K923166
1. Comparison with predicate: Toxoplasma gondii IgM Predicate Device: VIDAS® TOXO IgM

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Device Similarities
Characteristics	New DeviceBioPlex 2200 ToRC IgM	Predicate DevicebioMeriéux, Inc.,VIDAS® TOXO IgMK923166
Intended Use	Qualitative detection ofIgM antibodies toToxoplasma gondii (T.gondii), Rubella andCytomegalovirus (CMV)	Qualitative detection ofanti-Toxoplasma gondiiantibodies
Indications for Use	To be used as an aid inthe diagnosis of acurrent or recent T.gondii, Rubella and/orCMV infection	To be used as an aid inthe diagnosis of acute,recent, or reactivatedToxoplasma gondiiinfection
Measured Analyte	IgM antibodies toToxoplasma gondii (T.gondii), Rubella andCytomegalovirus(CMV)	Same
Signal Detection	Fluorescence	Same

Device Differences
Characteristics	New DeviceBioPlex 2200 ToRC IgM	Predicate DevicebioMerieux, Inc.,VIDAS® TOXO IgMK923166
Sample HandlingProcess	Automated	Manual
Assay Technology	Automated multiplexflow immunoassay	Automated enzymelinkedfluorescentimmunoassay (ELFA)
Solid Phase	Antigen-coatedparamagnetic	Antibody Coated SolidPhase Receptacle
Device Differences
Characteristics	New DeviceBioPlex 2200 ToRC IgM	Predicate DevicebioMeriéux, Inc.,VIDAS® TOXO IgMK923166
	microbeads	(SPR®)
Reagent IntegralStorage	On-board or inrefrigerator at 2-8°C	No reagent storage
Enzyme Conjugate	Phycoerythrinconjugated	Alkaline phosphataseconjugated
Unit of Measure	AI	RFV
Calibrator(s)	2 calibrator levels (soldseparately)	Single calibratorincluded with the kit
Reagent PackCalibrationFrequency	Every 30 days	Every 14 days
Control IntendedUse	Use as an assayedquality control tomonitor the overallperformance of theBioPlex 2200 Instrumentand BioPlex 2200 ToRCIgM Reagent Pack inthe clinical laboratory	Intended for use asassayed quality controlsamples to monitor theaccuracy and precisionof the assay
Instrumentation	Bio-Rad BioPlex 2200System	Vitek®ImmunoDiagnosticAssay System,VIDAS®System
Sample Matrix	Serum and Plasma(EDTA and Heparin)	Serum
Sample Size	3 µL	100 µL
Device Similarities
Characteristics	New DeviceBioPlex 2200 ToRCIgM	Predicate DeviceBioPlex 2200 Rubellaand CMV IgM KitK092587
Intended Use	Qualitative detection ofIgM antibodies toToxoplasma gondii (T.gondii), Rubella andCytomegalovirus(CMV)	Same
Indications forUse	To be used as an aid inthe diagnosis of acurrent or recent T.gondii, Rubella and/orCMV infection, inindividuals suspectedof having one of therespective diseasestates including womenof child bearing age.	Same
MeasuredAnalyte	IgM antibodies toToxoplasma gondii (T.gondii), Rubella andCytomegalovirus(CMV)	Same
AssayTechnology	Automated multiplexflow immunoassay	Same
Test Principle	Multiplexed flowimmunoassay	Same
Solid Phase	Antigen-coatedparamagneticmicrobeads	Same
SampleHandling/Process	Automated	Same
Reagent IntegralStorage	On-board or inrefrigerator at 2-8°C	Same
Signal Detection	Fluorescence	Same
	Device Similarities
Characteristics	New DeviceBioPlex 2200 ToRCIgM	Predicate DeviceBioPlex 2200 Rubellaand CMV IgM KitK092587
Unit of Measure	AI	Same
Reagent PackCalibrationFrequency	Every 30 days	Same
Control IntendedUse	Use as an assayedquality control tomonitor the overallperformance of theBioPlex 2200Instrument and BioPlex2200 ToRC IgMReagent Pack in theclinical laboratory	Same
Instrumentation	Bio-Rad BioPlex 2200System	Same

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Rubella and CMV IgM Predicate Device: BioPlex 2200 Rubella and CMV IgM Kit

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Device Differences
Characteristics	New DeviceBioPlex 2200 ToRCIgM	Predicate DeviceBioPlex 2200 Rubellaand CMV IgM KitK092587
Conjugate	Phycoerythrin-conjugated murinemonoclonal anti-humanIgM antibody andphycoerythrin-conjugated murinemonoclonal anti-humanFXIII antibody	Phycoerythrin-conjugated donkeypolyclonal anti-humanIgM antibody andphycoerythrin-conjugated murinemonoclonal anti-human FXIII antibody
Sample Matrix	Serum and Plasma	Serum, potassiumEDTA or Sodium

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Device Differences
Characteristics	New DeviceBioPlex 2200 ToRCIgM	Predicate DeviceBioPlex 2200 Rubellaand CMV IgM KitK092587
	(EDTA and Heparin)	Heparin plasma
Calibrator(s)	2 calibrator levels (soldseparately)	3 calibrator levels(sold separately)

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)

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, 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 then agitates the mixture at 37°C. In the second step, immobilized IgM is identified indirectly using a fluorescent 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 callbrator 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). Refer to the BioPlex 2200 System Operation Manual for more information about control beads. 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 in flow using a dual laser detector employing the same principles utilized by fluorescence activated cell sorters. Raw data are reported as relative fluorescent intensity (RFI).

Performance Characteristics (if/when applicable):

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1. Analytical performance:

a. Precision/Reproducibility:

Precision Studies:

Precision testing of the BioPlex 2200 ToRC IgM kit on the BioPlex 2200 instrument was performed in accordance with 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-day, and total reproducibility and the mean (AI), standard deviation (AI) and percent coefficient of variation (%CV) are summarized below. SD was evaluated for samples with AI values <0.8 and %CV was used for samples with AI values >0.8.

					Within Run		Between Run		Between Day		Total
Sample Type	T. gondii IgMPanel 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%
PotassiumEDTA	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%

BioPlex 2200 Toxo IgM – CLSI EP5-A3 Precision

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				Within Run		Between Run		Between-Day		Total
Sample Type	Rubella IgMPanel 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%
PotassiumEDTA	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%

BioPlex 2200 Rubella IgM – CLSI EP5-A3 Precision

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

			Within Run		Between Run		Between-Day		Total
Sample Type	CMV IgM PanelMembers	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%
PotassiumEDTA	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%

BioPlex 2200 CMV IgM – CLSI EP5-A3 Precision

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

Reproducibility Studies:

A reproducibility panel, consisting of six (6) panel members made using serum matrix and BioPlex ToRC IgM OC controls was prepared by Bio-Rad Laboratories. The serum sample panel includes 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 run x 5 days x 3 Sites = 120 replicates per panel member). The data were analyzed for withinrun (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. SD was evaluated for samples with AI values <0.8 and %CV was used for samples with AI values >0.8.

T. gondii IqM	Mean	Within Run		Between Run		Between Day		Between Site		Total
Panel Member	(AI)	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 Toxo IgM - CLSI EP15-A3 Reproducibility

BioPlex 2200 Rubella IgM - CLSI EP15-A3 Reproducibility

Rubella IgM	Mean	Within Run		Between Run		Between Day		Between Site		Total
Panel Member	(AI)	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 - CLSI EP15-A3 Reproducibility

CMV IgM PanelMember	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%

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

b. Linearity/assay reportable range:

Not Applicable.

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

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 and multianalyte T. gondii, Rubella CMV IgM calibrator is used to calibrate the T. gondii, Rubella and CMV IgM assays. The cut-off value and assignment of the calibrators are 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 negative control, multianalyte T. gondii, Rubella and CMV IgM. The positive controls contain known concentrations of human T. gondii, Rubella and CMV IgM, and are 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)
NegativeControl	0.0 - 0.5
Positive Control	2.0 - 2.6

Kit Stability

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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℃; 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℃), 60 days from first opening: Onboard Calibration Curve Stability, 30 days: Real Time Stability of calibrators and controls (2 to 8℃), 12 months; labeled as until expiration date; Calibrators and Controls Accelerated Stability (2 to 8℃), 18 months predicted. Calibrators and Controls Freeze-thaw (-20℃ or -70℃), 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℃) for up to 3 days and under refrigeration (2 - 8℃) for up to 7 days. For longer storage of samples, keep at -20°C or colder. Up to 5-freeze thaw cycles at -20℃ and -70℃ 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 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.

	T. gondii IgM				Rubella IgM				CMV IgM
Potential Cross Reactant	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

BioPlex 2200 ToRC IgM Cross Reactivity

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

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 kit according to CLSI EP7-A2 guideline.

No significant interference was observed in 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	≤ 6 mg/dL
Sodium Heparin	≤ 8000 units/dL
Lithium Heparin	≤ 8000 units/dL
EDTA (K2 and K3)	≤ 800 mg/dL

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

High dose hook effect: Not Applicable

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 samples for T. gondli, Rubella and CMV IgM, respectively, of which the serological status was determined from the predicate T. gondii, Rubella and CMV IgM assays. The assay will employ 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:

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.

MatrixComparison	N	BioPlexToRC IgMAssay	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
K3 EDTA vs.Serum	53	Rubella IgM	1.03(1.00 to 1.06)	-0.02(-0.08 to 0.05)	0.994
K3 EDTA vs.Serum	60	CMV IgM	1.01(0.97 to 1.05)	-0.02(-0.10 to 0.05)	0.989
LithiumHeparin 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
SodiumHeparin 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:

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

Not Applicable

b. Clinical Specificity: Not Applicable
c. Other clinical supportive data (when a. and b. are not applicable): Not Applicable
Clinical cut-off: 4. Not Applicable
ನ. 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. gondli, 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.

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

	T. gondii IgM		Rubella IgM		CMV IgM
Age	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%

*Since 2001 the incidence of Rubella in the US has been less than 10/1,000,000 population

Other Clinical Supportive Data: 6.

a. Method comparison with predicate device: Method comparison studies were performed following CLSI EP12-A2 guideline.

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

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 2129 prospective samples (~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 below.

Comparative Testing: Prospective

BioPlex 2200 ToRC IgM vs commercially available immunoassay for samples sent for T.
gondii, Rubella or CMV IgM testing

			BioPlex 2200 ToRC IgM Agreement
		Test Ordered		Pos (+)	Eqv	Neg (-)	Total	Pos (+)% Agreement95% CI	Neg (-)% Agreement95% CI
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
Commercially Available Immunoassay	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	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

10 samples were considered predive (7 positives and 3 equivocal). Of these 10 samples BioPlex Rubella (gM was negative for 5 of 10 and equivocal on 1 of 10. ** 2 out of the 16 CMV IgM pregnant women samples by predicate and negative by BioPex CMV (gM. 14 out of 16 were positive by predicate and BioPex results in these were as follows: 2 were negative and 8 were positive by Bioplex CMV (M. Of the 8 discrepart samples, 7 were negative and one equivoca by another FDA cleared device.

4 out of 36 CMV IgM test ordered samples were equive by BioPlex CMV IgM. 32 out of 36 were positive by predicate and BioPlex results in these were as follows: 1 was equive and 20 were positive by Bioplex CMV IgM. 13 of the 16 discrepant samples were confimed negative by another FDA cleared device

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

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 used for the comparative analysis. The characteristics of samples with presumptive positive status are shown below.

Comparative Testing: Retrospective

			BioPlex 2200 ToRC IgM vs commercially available immunoassay for T.
		gondii, Rubella or CMV IgM presumptive positive samples

			BioPlex 2200 ToRC IgM Agreement
		Presumptive Positive forT. gondii , Rubella or CMVIgM	Pos (+)	Eqv	Neg (-)	Total	Pos (+)% Agreement95% 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: b.

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.

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

CDC PanelT. gondiiIgM	BioPlex T. gondii IgM				PositiveAgreement	NegativeAgreement
	Pos (+)	Eq	Neg (-)	Total
Pos (+)	32	0	0	32	100.0%	100.0%
Neg (-)	0	0	65	65	89.3 to 100.0	94.4 to 100.0
Total	32	0	65	97

Characteristics of CDC T. gondii Reference Sera (N=97)

Seroconversion Testing: C.
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	VIDAS Toxo IgMIndex	BioPlex ToRC IgMAntibody Index (AI)
	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)
Antibody Systems	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)

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

Panel	Day	Rubella IgM
		BioPlex RC IgMAntibody Index (AI)	BioPlex ToRC IgMAntibody 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)

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

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

Panel	Day	CMV IgM
		BioPlex RC IgMAntibody Index (AI)	BioPlex ToRC IgMAntibody 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)

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

d. 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.

Sample ID	T. gondii IgM (AI) Before DTT	DTT Treatment Al(% 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%)

BioPlex ToRC IgM - T. gondii IgM Specificity

{25}------------------------------------------------

Sample ID	Rubella IgM (AI) Before DTT	DTT Treatment Al(% 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 – Rubella IgM Specificity

BioPlex ToRC IgM – CMV IgM Specificity

Sample ID	CMV IgM (Al) Before DTT	DTT Treatment Al(% 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%)

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.