BioPlex 2200 ToRC IgM, BioPlex 2200 ToRC IgM Calibrator Set, BioPlex 2200 ToRC IgM Control Set
Device Facts
| Record ID | K170509 |
|---|---|
| Device Name | BioPlex 2200 ToRC IgM, BioPlex 2200 ToRC IgM Calibrator Set, BioPlex 2200 ToRC IgM Control Set |
| Applicant | Bio-Rad Laboratories |
| Product Code | PUQ · Microbiology |
| Decision Date | May 19, 2017 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.3510 |
| Device Class | Class 2 |
Intended 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.
Indications for Use
Indicated for qualitative detection of IgM antibodies to Toxoplasma gondii, Rubella, and CMV in human serum and plasma to aid in diagnosis of current or recent infection in individuals suspected of having these diseases, including women of child-bearing age.
Regulatory Classification
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
Related Devices
- K092587 — BIOPLEX 2200 RUBELLA & CMV IGM KIT ON THE BIOPLEX 2200 MULTI ANALYTE DETECTION SYSTEM · Bio-Rad Laboratories · Dec 3, 2010
Submission Summary (Full Text)
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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 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.
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
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.
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: Multiplex<br>flow immunoassay,<br><i>T. gondii</i> , Rubella,<br>CMV IgM | Class II | 21 CFR § 866.3510 – Rubella virus<br>Serological Agents | Microbiology |
| JIT, Calibrator,<br>Secondary | Class II | 21 CFR §862.1150 –<br>Calibrator | Clinical Chemistry |
| JJY, Multi-Analyte<br>Controls, All Kinds<br>(Assayed) | Class I | 21 CFR §862.1660 –<br>Quality Control<br>Material (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.
1. 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 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.
- 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
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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.
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.
## Substantial Equivalence Information:
- 1. Predicate device name(s): BioPlex 2200 Rubella and CMV IgM Kit, K092587 bioMeriéux, Inc. VIDAS® TOXO IgM, K923166
- 2. Comparison with predicate: Toxoplasma gondii IgM Predicate Device: VIDAS® TOXO IgM
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| Device Similarities | | |
|---------------------|-------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------|
| Characteristics | New Device<br>BioPlex 2200 ToRC IgM | Predicate Device<br>bioMeriéux, Inc.,<br>VIDAS® TOXO IgM<br>K923166 |
| Intended Use | Qualitative detection of<br>IgM antibodies to<br>Toxoplasma gondii (T.<br>gondii), Rubella and<br>Cytomegalovirus (CMV) | Qualitative detection of<br>anti-Toxoplasma gondii<br>antibodies |
| Indications for Use | To be used as an aid in<br>the diagnosis of a<br>current or recent T.<br>gondii, Rubella and/or<br>CMV infection | To be used as an aid in<br>the diagnosis of acute,<br>recent, or reactivated<br>Toxoplasma gondii<br>infection |
| Measured Analyte | IgM antibodies to<br>Toxoplasma gondii (T.<br>gondii), Rubella and<br>Cytomegalovirus<br>(CMV) | Same |
| Signal Detection | Fluorescence | Same |
| Device Differences | | |
|------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------|
| Characteristics | New Device<br>BioPlex 2200 ToRC IgM | Predicate Device<br>bioMerieux, Inc.,<br>VIDAS® TOXO IgM<br>K923166 |
| Sample Handling<br>Process | Automated | Manual |
| Assay Technology | Automated multiplex<br>flow immunoassay | Automated enzyme<br>linked<br>fluorescent<br>immunoassay (ELFA) |
| Solid Phase | Antigen-coated<br>paramagnetic | Antibody Coated Solid<br>Phase Receptacle |
| Device Differences | | |
| Characteristics | New Device<br>BioPlex 2200 ToRC IgM | Predicate Device<br>bioMeriéux, Inc.,<br>VIDAS® TOXO IgM<br>K923166 |
| | microbeads | (SPR®) |
| Reagent Integral<br>Storage | On-board or in<br>refrigerator at 2-8°C | No reagent storage |
| Enzyme Conjugate | Phycoerythrin<br>conjugated | Alkaline phosphatase<br>conjugated |
| Unit of Measure | AI | RFV |
| Calibrator(s) | 2 calibrator levels (sold<br>separately) | Single calibrator<br>included with the kit |
| Reagent Pack<br>Calibration<br>Frequency | Every 30 days | Every 14 days |
| Control Intended<br>Use | Use as an assayed<br>quality control to<br>monitor the overall<br>performance of the<br>BioPlex 2200 Instrument<br>and BioPlex 2200 ToRC<br>IgM Reagent Pack in<br>the clinical laboratory | Intended for use as<br>assayed quality control<br>samples to monitor the<br>accuracy and precision<br>of the assay |
| Instrumentation | Bio-Rad BioPlex 2200<br>System | Vitek®<br>ImmunoDiagnostic<br>Assay System,VIDAS®<br>System |
| Sample Matrix | Serum and Plasma<br>(EDTA and Heparin) | Serum |
| Sample Size | 3 µL | 100 µL |
| Device Similarities | | |
| Characteristics | New Device<br>BioPlex 2200 ToRC<br>IgM | Predicate Device<br>BioPlex 2200 Rubella<br>and CMV IgM Kit<br>K092587 |
| Intended Use | Qualitative detection of<br>IgM antibodies to<br>Toxoplasma gondii (T.<br>gondii), Rubella and<br>Cytomegalovirus<br>(CMV) | Same |
| Indications for<br>Use | To be used as an aid in<br>the diagnosis of a<br>current or recent T.<br>gondii, Rubella and/or<br>CMV infection, in<br>individuals suspected<br>of having one of the<br>respective disease<br>states including women<br>of child bearing age. | Same |
| Measured<br>Analyte | IgM antibodies to<br>Toxoplasma gondii (T.<br>gondii), Rubella and<br>Cytomegalovirus<br>(CMV) | Same |
| Assay<br>Technology | Automated multiplex<br>flow immunoassay | Same |
| Test Principle | Multiplexed flow<br>immunoassay | Same |
| Solid Phase | Antigen-coated<br>paramagnetic<br>microbeads | Same |
| Sample<br>Handling/Process | Automated | Same |
| Reagent Integral<br>Storage | On-board or in<br>refrigerator at 2-8°C | Same |
| Signal Detection | Fluorescence | Same |
| | Device Similarities | |
| Characteristics | New Device<br>BioPlex 2200 ToRC<br>IgM | Predicate Device<br>BioPlex 2200 Rubella<br>and CMV IgM Kit<br>K092587 |
| Unit of Measure | AI | Same |
| Reagent Pack<br>Calibration<br>Frequency | Every 30 days | Same |
| Control Intended<br>Use | Use as an assayed<br>quality control to<br>monitor the overall<br>performance of the<br>BioPlex 2200<br>Instrument and BioPlex<br>2200 ToRC IgM<br>Reagent Pack in the<br>clinical laboratory | Same |
| Instrumentation | Bio-Rad BioPlex 2200<br>System | 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 Device<br>BioPlex 2200 ToRC<br>IgM | Predicate Device<br>BioPlex 2200 Rubella<br>and CMV IgM Kit<br>K092587 | | | | | | |
| Conjugate | Phycoerythrin-<br>conjugated murine<br>monoclonal anti-human<br>IgM antibody and<br>phycoerythrin-<br>conjugated murine<br>monoclonal anti-human<br>FXIII antibody | Phycoerythrin-<br>conjugated donkey<br>polyclonal anti-human<br>IgM antibody and<br>phycoerythrin-<br>conjugated murine<br>monoclonal anti-<br>human FXIII antibody | | | | | | |
| Sample Matrix | Serum and Plasma | Serum, potassium<br>EDTA or Sodium | | | | | | |
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| Device Differences | | |
|--------------------|------------------------------------------|------------------------------------------------------------------------|
| Characteristics | New Device<br>BioPlex 2200 ToRC<br>IgM | Predicate Device<br>BioPlex 2200 Rubella<br>and CMV IgM Kit<br>K092587 |
| | (EDTA and Heparin) | Heparin plasma |
| Calibrator(s) | 2 calibrator levels (sold<br>separately) | 3 calibrator levels<br>(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 IgM<br>Panel Members | N | Mean<br>(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<br>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 Toxo IgM – CLSI EP5-A3 Precision
{12}------------------------------------------------
| | | | | Within Run | | Between Run | | Between-Day | | Total | |
|-------------------|------------------------------|----|--------------|------------|-------|-------------|------|-------------|------|-------|-------|
| Sample Type | Rubella IgM<br>Panel Members | N | Mean<br>(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<br>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 Rubella IgM – CLSI EP5-A3 Precision
{13}------------------------------------------------
| | | | Within Run | | Between Run | | Between-Day | | Total | | |
|-------------------|--------------------------|----|--------------|-------|-------------|-------|-------------|-------|-------|-------|-------|
| Sample Type | CMV IgM Panel<br>Members | N | Mean<br>(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<br>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% |
## 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 Panel<br>Member | Mean<br>(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) |
|---------------------|------------|
| Negative<br>Control | 0.0 - 0.5 |
| Positive Control | 2.0 - 2.6 |
## Kit Stability
{16}------------------------------------------------
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.
| | <i>T. gondii</i> 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.
| Matrix<br>Comparison | N | BioPlex<br>ToRC IgM<br>Assay | Slope<br>(95% CI) | Intercept<br>(95% CI) | Correlation<br>(r) |
|---------------------------------|----|------------------------------|------------------------|--------------------------|--------------------|
| K3 EDTA vs.<br>Serum | 54 | <i>T. gondii</i> IgM | 1.02<br>(0.99 to 1.06) | -0.05<br>(-0.12 to 0.02) | 0.994 |
| K3 EDTA vs.<br>Serum | 53 | Rubella IgM | 1.03<br>(1.00 to 1.06) | -0.02<br>(-0.08 to 0.05) | 0.994 |
| K3 EDTA vs.<br>Serum | 60 | CMV IgM | 1.01<br>(0.97 to 1.05) | -0.02<br>(-0.10 to 0.05) | 0.989 |
| Lithium<br>Heparin vs.<br>Serum | 54 | <i>T. gondii</i> IgM | 1.02<br>(0.98 to 1.06) | -0.04<br>(-0.12 to 0.04) | 0.992 |
| | 51 | Rubella IgM | 1.03<br>(1.00 to 1.06) | -0.02<br>(-0.08 to 0.04) | 0.995 |
| | 60 | CMV IgM | 0.98<br>(0.93 to 1.02) | 0.00<br>(-0.09 to 0.09) | 0.985 |
| Sodium<br>Heparin vs.<br>Serum | 54 | <i>T. gondii</i> IgM | 1.03<br>(0.99 to 1.06) | -0.05<br>(-0.12 to 0.02) | 0.994 |
| | 51 | Rubella IgM | 1.01<br>(0.97 to 1.05) | -0.01<br>(-0.09 to 0.06) | 0.993 |
| | 59 | CMV IgM | 0.99<br>(0.96 to 1.03) | -0.04<br>(-0.11 to 0.04) | 0.991 |
#### 3. Clinical studies:
a. Clinical Sensitivity:
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## 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
| | <i>T. gondii</i> 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.
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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 (+)<br>% Agreement<br>95% CI | Neg (-)<br>% Agreement<br>95% CI |
| T. gondii IgM | Pregnant Women | Pos (+) | 0 | 0 | 0 | 0 | N/A | 98.0%<br>(196/200)<br>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%<br>(481/494)<br>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%<br>(198/198)<br>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%<br>(4/10)*<br>16.8 to 68.7% | 99.6%<br>(498/500)<br>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%<br>(8/16)**<br>28.0 to 72.0% | 100.0%<br>(183/183)<br>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%<br>(20/36)***<br>39.6 to 70.5% | 98.6%<br>(480/487)<br>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
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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 for<br><i>T. gondii</i> , Rubella or CMV<br>IgM | Pos (+…
