← Product Code [MVM](/submissions/IM/subpart-f%E2%80%94immunological-test-systems/MVM) · K130053 # BIOPLEX 2200 CELIAC IGA AND IGG KITS ON THE BIOPLEX 2200 SYSTEM, BIOPLEX 2200 CELIAC IGA AND IGG CALIBRATOR SETS, AND BI (K130053) _Bio-Rad Laboratories, Inc. · MVM · Sep 19, 2013 · Immunology · SESE_ **Canonical URL:** https://fda.innolitics.com/submissions/IM/subpart-f%E2%80%94immunological-test-systems/MVM/K130053 ## Device Facts - **Applicant:** Bio-Rad Laboratories, Inc. - **Product Code:** [MVM](/submissions/IM/subpart-f%E2%80%94immunological-test-systems/MVM.md) - **Decision Date:** Sep 19, 2013 - **Decision:** SESE - **Submission Type:** Traditional - **Regulation:** 21 CFR 866.5660 - **Device Class:** Class 2 - **Review Panel:** Immunology ## Intended Use The BioPlex 2200 Celiac IgA kit is an in vitro multiplex flow immunoassay intended for the semi-quantitative detection of IgA autoantibodies to deamidated gliadin peptide (DGP) and tissue Transglutaminase (tTG) in human serum. In conjunction with clinical findings and other diagnostic tests, the test system is used as an aid in the diagnosis of Celiac Disease (gluten-sensitive enteropathy). The BioPlex 2200 Celiac IgA kit is intended for use with the Bio-Rad BioPlex 2200 System. The BioPlex 2200 Celiac IgG kit is an in vitro multiplex flow immunoassay intended for the semi-quantitative detection of IgG autoantibodies to deamidated gliadin peptide (DGP) and tissue Transglutaminase (tTG) in human serum. In conjunction with clinical findings and other diagnostic tests, the test system is used as an aid in the diagnosis of Celiac Disease (gluten-sensitive enteropathy). The BioPlex 2200 Celiac IgG kit is intended for use with the Bio-Rad BioPlex 2200 System. ## Device Story The BioPlex 2200 Celiac IgA and IgG kits are multiplexed, bead-based immunoassays designed for use on the automated BioPlex 2200 System. The device uses dyed paramagnetic beads coated with recombinant tTG and DGP antigens. Patient serum is incubated with these beads; captured antibodies are detected using phycoerythrin-conjugated anti-human IgA or IgG. The system uses flow cytometry to identify bead populations and measure fluorescence intensity (RFI). Internal standard and serum verification beads monitor system performance and sample addition. Results are calculated via a 4-parameter logistic (4-PL) curve from a 5-point calibration. The device is used in clinical laboratories to aid in diagnosing Celiac disease by identifying specific autoantibodies. It provides semi-quantitative results in U/mL, which clinicians use alongside other diagnostic findings to support a Celiac diagnosis. ## Clinical Evidence Clinical study evaluated 319 specimens (156 biopsy-confirmed Celiac, 163 non-Celiac controls). Anti-tTG IgA sensitivity 94.9%, specificity 98.8%. Anti-DGP IgA sensitivity 87.2%, specificity 96.9%. Anti-tTG IgG sensitivity 44.2%, specificity 95.7%. Anti-DGP IgG sensitivity 84.6%, specificity 96.9%. Method comparison against predicate ELISAs showed high total agreement (94.5% for IgA, 89.5% for IgG). ## Technological Characteristics Multiplexed flow immunoassay using dyed paramagnetic microbeads coated with recombinant tTG and DGP antigens. Detection via phycoerythrin-conjugated anti-human IgA/IgG. Automated system (BioPlex 2200) uses fluorescence detection. Calibrated using 5-point curves (4-PL). Includes internal standard and serum verification beads. Preservatives: ProClin 300, sodium benzoate, sodium azide. ## Regulatory Identification A multiple autoantibodies immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoantibodies (antibodies produced against the body's own tissues) in serum and other body fluids. Measurement of multiple autoantibodies aids in the diagnosis of autoimmune disorders (disease produced when the body's own tissues are injured by autoantibodies). ## Predicate Devices - Quanta Lite™ h-tTG IgA (k011566) - Quanta Lite™ Gliadin IgA II (k052143) - Quanta Lite™ h-tTG IgG (k011570) - Quanta Lite™ Gliadin IgG II (k052142) ## Submission Summary (Full Text) > This content was OCRed from public FDA records by [Innolitics](https://innolitics.com). If you use, quote, summarize, crawl, or train on this content, cite Innolitics at https://innolitics.com. > > Innolitics is a medical-device software consultancy. We help companies design, build, and clear FDA-regulated software and AI/ML devices, including [a 510(k)](https://innolitics.com/services/510ks/), [a De Novo](https://innolitics.com/services/regulatory/), [a SaMD](https://innolitics.com/services/end-to-end-samd/), [an AI/ML medical device](https://innolitics.com/services/medical-imaging-ai-development/), or [an FDA regulatory strategy](https://innolitics.com/services/regulatory/). {0}------------------------------------------------ K.130053 SEP 1 9 2013 # BioPlex® 2200 Celiac IgA and IgG 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 of 510(k) safety and effectiveness information provides detail as a basis for a determination of substantial equivalence for the BioPlex® 2200 Celiac IgA and IgG kits. ### 510(k) Number: K130053 ### Summary Preparation Date: August 14, 2013 August 14, 2013 ### Applicant: Bio-Rad Laboratories ### Contact: Juang Wang Regulatory Affairs Representative Purpose for Submission: New Device ### Measurand: IgA and IgG antibodies to tissue Transglutaminase (tTG) IgA and IgG antibodies to deamidated gliadin peptide (DGP) ### Type of Test: Semi-Quantitative multiplexed flow, bead-based immunoassay ### Proprietary and Established Names: BioPlex® 2200 Celiac IgA kit BioPlex® 2200 Celiac IgG kit BioPlex® 2200 Celiac IgA Calibrator Set BioPlex® 2200 Celiac IgA Control Set BioPlex® 2200 Celiac IgG Calibrator Set BioPlex® 2200 Celiac IgG Control Set {1}------------------------------------------------ ### Regulatory Information: | Product Code | Classification | Regulation Section | Panel | |--------------------------------------------------------------------|----------------|-----------------------------------------------------------------------------------|----------------------------| | Autoantibodies,
endomysial(tissue
transglutaminase)
(MVM) | Class II | 21 CFR § 866.5660,
Multiple autoantibodies
immunological test system. | Immunology (82) | | Antibodies, Gliadin
(MST) | Class II | 21 CFR § 866.5750,
Radioallergosorbent
(RAST) immunological test
system. | Immunology (82) | | Calibrator, Multi-Analyte
Mixture
(JIX) | Class II | 21 CFR § 862.1150 -
Calibrator | Clinical Chemistry
(75) | | Multi-Analyte Controls
All kinds(assayed)
(JJY) | Class I | 21 CFR § 862.1660 -
Quality Control Material
(Assayed and Unassayed) | Clinical Chemistry
(75) | ### Intended Use: - l . Intended use(s): BioPlex® 2200 Celiac IgA Kit The BioPlex 2200 Celiac IgA kit is an in vitro multiplex flow immunoassay intended for the semi-quantitative detection of IgA autoantibodies to deamidated gliadin peptide (DGP) and tissue Transglutaminase (tTG) in human serum. In conjunction with clinical findings and other diagnostic tests, the test system is used as an aid in the diagnosis of Celiac Disease (gluten-sensitive enteropathy). The BioPlex 2200 Celiac IgA kit is intended for use with the Bio-Rad BioPlex 2200 System. # BioPlex®2200 Celiac IgG Kit The BioPlex 2200 Celiac IgG kit is an in vitro multiplex flow immunoassay intended for the semi-quantitative detection of IgG autoantibodies to deamidated gliadin peptide (DGP) and tissue Transglutaminase (tTG) in human serum. In conjunction with clinical findings and other diagnostic tests, the test system is used as an aid in the diagnosis of Celiac Disease (gluten-sensitive enteropathy). The BioPlex 2200 Celiac IgG kit is intended for use with the Bio-Rad BioPlex 2200 System. {2}------------------------------------------------ # BioPlex® 2200 Celiac IgA Calibrator Set The BioPlex 2200 Celiac IgA Calibrator Set is intended for the calibration of the BioPlex 2200 Celiac IgA Reagent Pack. ### BioPlex® 2200 Celiac IgG Calibrator Set The BioPlex 2200 Celiac IgG Calibrator Set is intended for the calibration of the BioPlex 2200 Celiac IgG Reagent Pack. # BioPlex® 2200 Celiac IgA Control Set The BioPlex 2200 Celiac IgA Control Set is intended for use as an assayed quality control to monitor the overall performance of the BioPlex 2200 Instrument and BioPlex 2200 Celiac IgA Reagent Pack in the clinical laboratory. The performance of the BioPlex Celiac IgA Control Set has not been established with any other antitissue Transglutaminase (tTG) and anti-Deamidated Gliadin Peptide IgA assays. ### BioPlex® 2200 Celiac IgG Control Set The BioPlex 2200 Celiac IgG Control Set is intended for use as an assayed quality control to monitor the overall performance of the BioPlex 2200 Instrument and BioPlex 2200 Celiac IgG Reagent Pack in the clinical laboratory. The performance of the BioPlex Celiac IgG Control Set has not been established with any other antitissue Transglutaminase (tTG) and anti-Deamidated Gliadin Peptide IgG assays. - 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 ### Device Description: BioPlex® 2200 Celiac IgA and IgG kits include the following components: - . One (1) 10 mL vial of Bead Set, containing dyed beads coated with recombinant antigens; an Internal Standard bead (ISB), a Serum Verification bead (SVB) and IgA Verification Bead (AVB) (in Celiac IgA only), in MOPS (3-[N-Morpholino] propanesulfonic acid) buffer supplemented with Glycerol and protein stabilizer (bovine and porcine). ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) are added as preservatives. {3}------------------------------------------------ - One (1) 5 mL vial of Conjugate, containing phycoerythrin conjugated murine . monoclonal anti-human IgA or IgG and phycoerythrin conjugated sheep anti-human . FXIII in MOPS (3-N-Morpholino] propanesulfonic acid) buffer supplemented with bovine protein stabilizers. ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) are added as preservatives. - . One (1) 10 mL vial of Sample Diluent, containing bovine and murine protein stabilizers in triethanolamine buffer. ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) are added as preservatives. BioPlex 2200 Celiac IgA and IgG Calibrator Sets contain nine (9) 0.5 mL vials of human antibodies to tTG and DGP in a buffer supplemented with protein stabilizer (porcine for IgA and porcine/human for IgG) with ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) as preservatives. BioPlex 2200 Celiac IgA and IgG Control Sets contain four (4) 1.5 mL vials of Positive Controls of human antibodies to tTG or DGP and two vials of Negative Controls in a human serum matrix made from defibrinated plasma; and, in a human serum matrix made from defibrinated plasma with ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) as preservatives. Additional materials required but not supplied include BioPlex 2200 Sheath Fluid containing Phosphate Buffered Saline (PBS) with ProClin 300 (<0.03%), and sodium azide (<0.1%) as preservatives; and BioPlex 2200 Wash Solution containing Phosphate Buffered Saline (PBS) and Tween 20 with ProClin 300 (≤0.03%) and sodium azide (<0.1%) as preservatives. ### Substantial Equivalence Information: - l. Predicate device name(s): INOVA QUANTA Lite h-tTG IgA INOVA QUANTA Lite h-tTG IgG INOVA QUANTA Lite Gliadin IgA II INOVA QUANTA Lite Gliadin IgG II - 2. Predicate 510(k) number(s): K011566, K011570, K052143, K052142 {4}------------------------------------------------ # 3. Comparison with predicate: | | Similarities | | | |------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Item | Device:
BioPlex 2200 Celiac IgA and IgG Kits | Predicate:
INOVA QUANTA Lite h-tTG IgA and IgG Kits | Predicate:
INOVA QUANTA Lite Gliadin IgA II and IgG II Kits | | Intended Use | Semi-quantitative detection of IgA and IgG autoantibodies to deamidated gliadin peptide (DGP) and tissue Transglutaminase (tTG) in human serum. In conjunction with clinical findings and other diagnostic tests, the test system is used as an aid in the diagnosis of Celiac Disease (gluten-sensitive enteropathy). | Semi-quantitative detection of IgA and IgG autoantibodies to tissue Transglutaminase (endomysium) in human serum. Detection of these antibodies is an aid in diagnosis of certain gluten sensitive enteropathies such as celiac disease and dermatitis herpetiformis. | Semi-quantitative detection of gliadin IgA and IgG antibodies in human serum. The presence of gliadin antibodies can be used in conjunction with clinical findings and other laboratory tests to aid in the diagnosis of celiac disease. | | Sample Type | Serum | Serum | Serum | | Assay Type | Semi-quantitative | Same | Same | | Analyte Detected | Human IgA or IgG antibodies to human tissue transglutaminase | Same | Not Applicable | | | Human IgA or IgG antibodies to deamidated gliadin peptide | Not Applicable | Same | | Item | Differences | | | | | Device:
BioPlex 2200 Celiac IgA and
IgG Kits | Predicate:
INOVA QUANTA Lite
h-tTG IgA and IgG
Kits | Predicate:
INOVA QUANTA Lite
Gliadin IgA II and IgG
II Kits | | Assay Technology | Automated multiplex flow
immunoassay | Manual, microtiter
plate format,
Enzyme-linked
Immunosorbent assay
(ELISA) | Manual, microtiter
plate format,
Enzyme-linked
Immunosorbent assay
(ELISA) | | Capture Antigen | Recombinant tTG protein
and DGP antigen | Native human tissue
transglutaminase (h-
tTG) | Synthetic,
deamidated peptide | | Conjugate | Phycoerythrin conjugated | Goat anti-human IgG | Goat anti-human IgG | · {5}------------------------------------------------ | Differences | | | | | | | | | |-----------------------------|--------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------|-------------------------------------------------------------------------|--|--|--|--|--| | ltem | Device:
BioPlex 2200 Celiac IgA and
IgG Kits | Predicate:
INOVA QUANTA Lite
h-tTG IgA and IgG
Kits | Predicate:
INOVA QUANTA Lite
Gliadin IgA II and IgG
II Kits | | | | | | | Antibody | murine monoclonal anti-
human IgA or IgG | or IgA HRP-
conjugated antibody
solution | or IgA HRP-
conjugated antibody
solution | | | | | | | Substrate | None | Tetramethylbenzidine
(TMB) Chromogen | Tetramethylbenzidine
(TMB) Chromogen | | | | | | | Signal Detection | Fluorescence | Color, read at 450nm | Color, read at 450nm | | | | | | | Solid Phase | Antigen-coated paramagnetic
microbead reagent | Antigen-coated solid
phase microtiter
wells | Antigen-coated solid
phase microtiter wells | | | | | | | Calibrator(s) | 5 levels of Calibrator for both
IgA and IgG | 1 level of Low
Positive control | l level of Low
Positive control | | | | | | | Control | One Negative and one
Positive Control | One Negative, one
low Positive, and one
high Positive
Controls | One Negative, one
low Positive, and one
high Positive
Controls | | | | | | | Assay range | Anti-tTG:
IgA: 0.5 to 250 U/mL
lgG: 0.8 to 250 U/mL
Anti-DGP:
IgA: 0.2 to 250 U/mL
IgG: 0.4 to 250 U/mL | Not Applicable | Not Applicable | | | | | | | Calibrators and
Controls | Sold Separately | Kit components | Kit components | | | | | | | Quantitation | Results are determined from
a standard calibration curve
utilizing a 4-PL (4-Parameter
Logistic) curve fitting. | One point calculation
from the OD of the
low positive control. | One point calculation
from the OD of the
low positive control. | | | | | | | Instrumentation | Bio-Rad BioPlex 2200
system | Spectrophotometer | Spectrophotometer | | | | | | ## Standard/Guidance Document Referenced (if applicable): CEN 13640:2002, Stability Testing of In Vitro Diagnostic Reagents EP05-A2, Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline, Second Edition. EP06-A, Evaluation of Linearity of Quantitative Measurement, Approved Guideline, Second Edition. EP07-A2, Interference Testing in Clinical Chemistry, Approved Guideline, Second Edition {6}------------------------------------------------ EP09-A2IR, Method Comparison and Bias Estimation Using Patient Samples: Approved Guideline, Second Edition (Interim Revision). EP12-A2, User Protocol for Evaluation of Oualitative test Performance, Approved Guideline, Second Edition. EP14-A2, Evaluation of Matrix Effects, Approved Guideline, Second Edition EP15-A2, User Verification of Performance for Precision and Trueness, Approved Guideline, Second Edition. EP17-A, Protocols for Determination of Limits of Detection and Limits of Quantification. Approved Guideline. ### Test Principle: The BioPlex 2200 Celiac IgA and IgG kits use a multiplexed micro particle bead-based immunoassay for the semi-quantitative detection of IgA and IgG antibodies to tissue Transglutaminase and deamidated gliadin peptide in human serum using the Luminex flow cytometry technology. In the BioPlex Celiac IgA and IgG assays, two (2) different populations of dyed beads are coated with antigens associated with celiac disease (recombinant antigens). The BioPlex 2200 System combines an aliquot of patient sample, sample diluent, and bead set reagent into a reaction vessel. The mixture is incubated at 37°C. After a wash cycle, anti-human IgA or IgG antibody conjugated to phycoerythrin (PE) is added to the dved beads and this mixture is incubated at 37°C. The excess conjugate is removed in another wash cycle, and the beads are re-suspended in wash buffer. The bead mixture then passes through the detector. The identity of the dyed beads is determined by the fluorescence of the dyes, and the amount of antibody captured by the antigen is determined by the fluorescence of the attached PE. Raw data is calculated in relative fluorescence intensity (RFI). Two additional dyed beads, Internal Standard Bead (ISB) and Serum Verification Bead (SVB) are present in each reaction mixture to verify detector response and the addition of serum to the reaction vessel. An additional control bead, IgA Verification Bead (AVB) has been included for the Celiac IgA kit to flag results for samples with deficient IgA levels. Refer to the BioPlex 2200 System Operation Manual for more information on internal quality control beads. The system is calibrated using a set of nine (9) distinct calibrator vials, supplied separately by Bio-Rad Laboratories. For anti-tTG, five (5) vials, representing five (5) different antibody concentrations, are used for semi-quantitative calibration for anti-tTG IgA and IgG. For anti-DGP, five (5) vials, representing five (5) different antibody concentrations, are used for semiquantitative calibration for anti-DGP IgA and IgG. The result for each of these antibodies is expressed in Units/mL (U/mL). ### Performance Characteristics (if/when applicable): - 1. Analytical performance: - a. Precision/Reproducibility: {7}------------------------------------------------ Precision testing of the BioPlex® 2200 Celiac IgA and IgG kits on the BioPlex® 2200 instrument was performed in accordance with CLSI EP5-A2. A serum panel consisting of samples spanning the measuring range was assayed in replicate twice daily over 20 days (n=80). One positive and one negative control were included. The data were analyzed for within-run, between-run, between-day, and total precision and the mean U/mL, standard deviation and percent coefficient of variation are summarized below: | Sample Type | N | Mean U/mL | Within Run | | Between Run | | Between Day | | Total Precision | | |------------------|----|-----------|------------|------|-------------|------|-------------|------|-----------------|-------| | | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | High Negative | 80 | 6.3 | 0.4 | 6.2% | 0.2 | 3.3% | 0.4 | 6.3% | 0.6 | 9.4% | | High Negative | 80 | 6.2 | 0.4 | 6.1% | 0.0 | 0.0% | 0.4 | 5.8% | 0.5 | 8.4% | | Near Cutoff | 80 | 12.5 | 0.9 | 6.9% | 0.0 | 0.0% | 1.1 | 8.4% | 1.4 | 10.9% | | Near Cutoff | 80 | 15.7 | 0.8 | 5.2% | 0.7 | 4.4% | 0.8 | 5.0% | 1.3 | 8.5% | | Low Positive | 80 | 36.5 | 2.3 | 6.4% | 1.2 | 3.4% | 2.6 | 7.1% | 3.7 | 10.2% | | Low Positive | 80 | 38.4 | 2.2 | 5.8% | 0.0 | 0.0% | 3.0 | 7.8% | 3.7 | 9.7% | | Mid Positive | 80 | 108.4 | 6.3 | 5.8% | 3.4 | 3.1% | 7.9 | 7.3% | 10.7 | 9.9% | | Mid Positive | 80 | 94.8 | 6.5 | 6.8% | 0.0 | 0.0% | 5.3 | 5.6% | 8.4 | 8.9% | | High Positive | 80 | 210.9 | 10.9 | 5.2% | 0.0 | 0.0% | 11.8 | 5.6% | 16.1 | 7.6% | | High Positive | 80 | 227.7 | 8.2 | 3.6% | 5.5 | 2.4% | 9.0 | 3.9% | 13.3 | 5.8% | | Negative Control | 80 | 1.3 | 0.1 | 5.6% | 0.0 | 0.0% | 0.1 | 5.1% | 0.1 | 7.5% | | Positive Control | 80 | 65.7 | 2.4 | 3.7% | 1.1 | 1.7% | 1.6 | 2.5% | 3.1 | 4.7% | # BioPlex® 2200 Celiac IgA – Anti-fTG ### BioPlex® 2200 Celiac IgA - Anti-DGP | Sample Type | N | Mean
U/mL | Within Run | | Between Run | | Between Day | | Total Precision | | |------------------|----|--------------|------------|------|-------------|------|-------------|------|-----------------|-------| | | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | High Negative | 80 | 8.3 | 0.5 | 6.4% | 0.0 | 0.0% | 0.7 | 8.9% | 0.9 | 11.0% | | High Negative | 80 | 4.1 | 0.3 | 6.7% | 0.3 | 6.5% | 0.2 | 4.6% | 0.4 | 10.4% | | Near Cutoff | 80 | 16.1 | 0.7 | 4.6% | 0.3 | 2.0% | 0.7 | 4.2% | 1.1 | 6.5% | | Near Cutoff | 80 | 15.3 | 0.9 | 6.0% | 0.0 | 0.0% | 0.5 | 3.3% | 1.1 | 6.8% | | Low Positive | 80 | 34.5 | 1.8 | 5.1% | 0.0 | 0.0% | 1.0 | 2.9% | 2.0 | 5.9% | | Low Positive | 80 | 33.9 | 2.0 | 5.8% | 0.7 | 1.9% | 2.1 | 6.3% | 3.0 | 8.8% | | Mid Positive | 80 | 96.8 | 6.1 | 6.3% | 0.0 | 0.0% | 2.8 | 2.9% | 6.7 | 6.9% | | Mid Positive | 80 | 98.5 | 4.6 | 4.7% | 0.0 | 0.0% | 5.7 | 5.8% | 7.3 | 7.5% | | High Positive | 80 | 185.5 | 8.4 | 4.5% | 0.0 | 0.0% | 12.2 | 6.6% | 14.8 | 8.0% | | High Positive | 80 | 200.3 | 9.3 | 4.6% | 0.0 | 0.0% | 10.9 | 5.5% | 14.3 | 7.2% | | Negative Control | 80 | 2.0 | 0.2 | 7.5% | 0.0 | 1.7% | 0.1 | 6.8% | 0.2 | 10.3% | | Positive Control | 80 | 60.5 | 2.0 | 3.4% | 0.8 | 1.3% | 2.5 | 4.1% | 3.3 | 5.4% | ### BioPlex® 2200 Celiac IgG - Anti-tTG | Sample Type | N | Mean
U/mL | Within Run | | Between Run | | Between Day | | Total Precision | | |---------------|----|--------------|------------|------|-------------|------|-------------|-------|-----------------|-------| | High Negative | 80 | 7.7 | 0.5 | 6.2% | 0.3 | 3.3% | 0.7 | 9.4% | 0.9 | 11.7% | | High Negative | 80 | 9.3 | 0.5 | 5.6% | 0.5 | 5.7% | 1.2 | 13.2% | 1.4 | 15.4% | | Near Cutoff | 80 | 14.6 | 0.9 | 5.8% | 0.4 | 2.6% | 1.2 | 8.4% | 1.5 | 10.5% | Page 8 of 21 {8}------------------------------------------------ | Near Cutoff | 80 | 15.7 | 1.3 | 8.1% | 0.0 | 0.0% | 0.8 | 4.9% | 1.5 | 9.5% | |------------------|----|-------|------|------|-----|------|------|------|------|------| | Low Positive | 80 | 33.3 | 1.4 | 4.1% | 1.3 | 3.9% | 1.5 | 4.5% | 2.4 | 7.2% | | Low Positive | 80 | 34.0 | 1.3 | 3.8% | 0.0 | 0.0% | 1.5 | 4.4% | 2.0 | 5.8% | | Mid Positive | 80 | 101.9 | 3.6 | 3.5% | 2.1 | 2.1% | 4.4 | 4.3% | 6.1 | 5.9% | | Mid Positive | 80 | 133.3 | 4.9 | 3.6% | 0.3 | 0.2% | 6.4 | 4.8% | 8.1 | 6.1% | | High Positive | 80 | 172.3 | 6.3 | 3.7% | 0.0 | 0.0% | 5.8 | 3.4% | 8.5 | 5.0% | | High Positive | 80 | 207.9 | 11.0 | 5.3% | 0.0 | 0.0% | 16.0 | 7.7% | 19.5 | 9.4% | | Negative Control | 80 | 1.9 | 0.08 | 4.5% | 0.1 | 3.3% | 0.1 | 6.1% | 0.2 | 8.3% | | Positive Control | 80 | 55.6 | 1.5 | 2.8% | 0.9 | 1.7% | 2.1 | 3.8% | 2.8 | 5.0% | ### BioPlex® 2200 Celiac IgG – Anti-DGP | Sample Type | | Mean | | Within Run | Between Run | | Between Day | | Total Precision | | |------------------|----|--------|------|------------|-------------|------|-------------|------|-----------------|-------| | | N | U/mL | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | High Negative | 80 | 10.1 | 0.4 | 4.0% | 0.2 | 2.2% | 0.4 | 3.9% | 0.6 | 6.0% | | High Negative | 80 | 11.6 | 0.6 | 5.0% | 0.6 | 4.8% | 0.8 | 6.8% | 1.1 | 9.7% | | Near Cutoff | 80 | 15.0 | 0.4 | 2.4% | 0.3 | 2.3% | 0.6 | 3.9% | 0.8 | 5.1% | | Near Cutoff | 80 | 16.6 | 0.8 | 4.9% | 0.5 | 3.1% | 0.9 | 5.3% | 1.3 | 7.8% | | Low Positive | 80 | 32.7 | 1.7 | 5.3% | 0.0 | 0.0% | 2.3 | 7.0% | 2.9 | 8.8% | | Low Positive | 80 | 33.2 | 1.1 | 3.3% | 0.0 | 0.0% | 0.9 | 2.6% | 1.4 | 4.2% | | Mid Positive | 80 | 89.7 | 4.1 | 4.6% | 2.6 | 2.9% | 6.1 | 6.8% | 7.8 | 8.7% | | Mid Positive | 80 | ਰੇੜ 'ਚ | 4.1 | 4.4% | 0.0 | 0.0% | 4.5 | 4.8% | 6.1 | 6.5% | | High Positive | 80 | 164.6 | 4.8 | 2.9% | 3.0 | 1.8% | 4.8 | 2.9% | 7.4 | 4.5% | | High Positive | 80 | 194.6 | ರಿ.4 | 4.8% | 2.9 | 1.5% | 10.6 | 5.5% | 14.5 | 7.4% | | Negative Control | 80 | 2.8 | 0.3 | 10.2% | 0.0 | 0.0% | 0.1 | 4.5% | 0.3 | 11.2% | | Positive Control | 80 | 57.6 | 1.6 | 2.7% | 1.4 | 2.4% | 2.2 | 3.9% | 3.0 | 5.3% | ### CLSI EP15-A2 Reproducibility Precision and reproducibility was also evaluated in accordance with CLSI EP15-A2 guideline "User Verification of Performance for Precision and Trueness, Vol 25, No 17". A different serum panel consisting of samples spanning the measuring range were assayed in 4 replicates per run, one run per day over 5 days (n=20). One positive and one negative control were included. The data were analyzed for within-run, between run (day), and total precision and the mean U/mL, standard deviation and percent coefficient of variation are summarized below: | | | Grand | Within Run | | | Between Run
(Day) | | Total Precision | |--------------|----|----------------|------------|------|------|----------------------|--------|-----------------| | Panel Member | N | Mean
(U/mL) | SD | %CV | SD | %CV | SD | %CV | | Negative | 20 | 6.1 | 0.27 | 4.4% | 0.00 | 0.0% | 0.27 | 4.4% | | Negative | 20 | 6.3 | 0.26 | 4.1% | 0.16 | 2.5% | 0.30 | 4.8% | | Near Cutoff | 20 | 12.2 | 0.47 | 3.8% | 0.29 | 2.3% | 0.55 | 4.5% | | Near Cutoff | 20 | 15.7 | 0.67 | 4.3% | 0.27 | 1.7% | 0.72 | 4.6% | | Low Positive | 20 | 38.5 | 2.25 | 5.8% | 0.75 | 2.0% | 2.37 · | 6.2% | | Low Positive | 20 | 40.1 | 3.02 | 7.5% | 0.00 | 0.0% | 3.02 | 7.5% | ### BioPlex® 2200 Celiac IgA - Anti-tTG IgA Page 9 of 21 {9}------------------------------------------------ | | | Grand
| Within Run | | | Between Run
(Day) | Total Precision | | |------------------|----|----------------|------------|------|------|----------------------|-----------------|------| | Panel Member | N | Mean
(U/mL) | SD | %CV | SD | %CV | SD | %CV | | Medium Positive | 20 | 84.9 | 3.86 | 4.6% | 0.48 | 0.6% | 3.89 | 4.6% | | Medium Positive | 20 | 111.9 | 4.39 | 3.9% | 5.01 | 4.5% | 6.66 | 6.0% | | High Positive | 20 | 207.9 | 9.67 | 4.6% | 0.00 | 0.0% | 9.67 | 4.6% | | High Positive | 20 | 219.1 | 7.88 | 3.6% | 3.84 | 1.8% | 8.77 | 4.0% | | Negative Control | 20 | 1.2 | 0.06 | 4.8% | 0.00 | 0.0% | 0.06 | 4.8% | | Positive Control | 20 | 67.4 | 4.02 | 6.0% | 0.00 | 0.0% | 4.02 | 6.0% | # BioPlex® 2200 Celiac IgA - Anti-DGP IgA . . - | BioPlex® 2200 Celiac IgA – Anti-DGP IgA | | | | | | | | | |-----------------------------------------|----|----------------|------------|------|------|----------------------|-----------------|------| | | | Grand | Within Run | | | Between Run
(Day) | Total Precision | | | Panel Member | N | Mean
(U/mL) | SD | %CV | SD | %CV | SD | %CV | | Negative | 20 | 4.3 | 0.15 | 3.6% | 0.00 | 0.0% | 0.15 | 3.6% | | Negative | 20 | 8.0 | 0.37 | 4.7% | 0.00 | 0.0% | 0.37 | 4.7% | | Near Cutoff | 20 | 15.4 | 0.39 | 2.5% | 0.11 | 0.7% | 0.41 | 2.6% | | Near Cutoff | 20 | 16.4 | 0.67 | 4.1% | 0.00 | 0.0% | 0.67 | 4.1% | | Low Positive | 20 | 32.4 | 1.25 | 3.8% | 0.00 | 0.0% | 1.25 | 3.8% | | Low Positive | 20 | 33.8 | 1.32 | 3.9% | 0.00 | 0.0% | 1.32 | 3.9% | | Medium Positive | 20 | 91.3 | 3.00 | 3.3% | 0.95 | 1.0% | 3.15 | 3.4% | | Medium Positive | 20 | 93.1 | 4.07 | 4.4% | 2.70 | 2.9% | 4.89 | 5.2% | | High Positive | 20 | 169.2 | 8.61 | 5.1% | 0.00 | 0.0% | 8.61 | 5.1% | | High Positive | 20 | 187.8 | 6.41 | 3.4% | 0.00 | 0.0% | 6.41 | 3.4% | | Negative Control | 20 | 2.0 | 0.11 | 5.6% | 0.06 | 3.0% | 0.13 | 6.3% | | Positive Control | 20 | 55.4 | 2.27 | 4.1% | 0.52 | 0.9% | 2.33 | 4.2% | # BioPlex® 2200 Celiac IgG - Anti-tTG IgG | Panel Member | N | Grand Mean (U/mL) | Within Run | | Between Run (Day) | | Total Precision | | |------------------|----|-------------------|------------|------|-------------------|------|-----------------|------| | | | | | | | | | | | | | | SD | %CV | SD | %CV | SD | %CV | | Negative | 20 | 7.1 | 0.23 | 3.2% | 0.16 | 2.3% | 0.28 | 3.9% | | Negative | 20 | 7.2 | 0.27 | 3.8% | 0.21 | 2.9% | 0.34 | 4.8% | | Near Cutoff | 20 | 12.7 | 0.50 | 3.9% | 0.24 | 1.9% | 0.55 | 4.4% | | Near Cutoff | 20 | 14.8 | 0.66 | 4.5% | 0.26 | 1.7% | 0.71 | 4.8% | | Low Positive | 20 | 30.2 | 0.87 | 2.9% | 0.47 | 1.5% | 0.99 | 3.3% | | Low Positive | 20 | 31.4 | 0.85 | 2.7% | 0.60 | 1.9% | 1.04 | 3.3% | | Medium Positive | 20 | 94.2 | 2.93 | 3.1% | 2.70 | 2.9% | 3.99 | 4.2% | | Medium Positive | 20 | 123.2 | 5.72 | 4.6% | 2.32 | 1.9% | 6.18 | 5.0% | | High Positive | 20 | 164.1 | 8.81 | 5.4% | 5.46 | 3.3% | 10.36 | 6.3% | | High Positive | 20 | 189.7 | 8.58 | 4.5% | 4.75 | 2.5% | 9.81 | 5.2% | | Negative Control | 20 | 1.8 | 0.18 | 9.8% | 0.00 | 0.0% | 0.18 | 9.8% | | Positive Control | 20 | 51.5 | 1.44 | 2.8% | 0.53 | 1.0% | 1.53 | 3.0% | - {10}------------------------------------------------ | | | Grand | Within Run | | | Between Run
(Day) | Total Precision | | |------------------|----|----------------|------------|------|------|----------------------|-----------------|------| | Panel Member | N | Mean
(U/mL) | SD | %CV | SD | %CV | SD | %CV | | Negative | 20 | 9.7 | 0.25 | 2.5% | 0.34 | 3.5% | 0.42 | 4.3% | | Negative | 20 | 11.0 | 0.37 | 3.4% | 0.00 | 0.0% | 0.37 | 3.4% | | Near Cutoff | 20 | 14.3 | 0.52 | 3.6% | 0.26 | 1.8% | 0.58 | 4.0% | | Near Cutoff | 20 | 15.9 | 0.52 | 3.3% | 0.29 | 1.8% | 0.59 | 3.7% | | Low Positive | 20 | 30.6 | 1.01 | 3.3% | 0.31 | 1.0% | 1.06 | 3.5% | | Low Positive | 20 | 32.5 | 1.45 | 4.5% | 0.71 | 2.2% | 1.61 | 5.0% | | Medium Positive | 20 | 88.8 | 2.65 | 3.0% | 0.64 | 0.7% | 2.72 | 3.1% | | Medium Positive | 20 | 91.4 | 3.59 | 3.9% | 3.36 | 3.7% | 4.92 | 5.4% | | High Positive | 20 | 172.5 | 4.57 | 2.7% | 2.45 | 1.4% | 5.19 | 3.0% | | High Positive | 20 | 202.8 | 5.62 | 2.8% | 4.86 | 2.4% | 7.43 | 3.7% | | Negative Control | 20 | 2.7 | 0.23 | 8.6% | 0.01 | 0.3% | 0.23 | 8.6% | | Positive Control | 20 | 54.1 | 1.36 | 2.5% | 0.65 | 1.2% | 1.51 | 2.8% | BioPlex® 2200 Celiac IgG – Anti-DGP IgG ### Lot-to-lot Reproducibility The lot-to-lot reproducibility study was conducted to evaluate the variation among the lots of the reagent kit. Serum samples spanning the assay range were tested with three reagent lots on three BioPlex 2200 instruments in replicates of 10 for two runs. Each lot mean (U/mL) was calculated using data 60 points for each patient serum sample (ten replicates, three instruments, two runs per instrument). The lot to lot grand mean (U/mL), standard deviation and %CV of the anti-DGP and anti-tTG IgA samples were calculated for each of the samples. | | | | Within Run | | Between Run | | Between Instrument/
Operator | | Between Lot | | Total | | |-----------|--------------|------|------------|-------|-------------|------|---------------------------------|-------|-------------|-------|-------|--| | Sample ID | Mean
U/mL | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | | SCePr14 | 7.2 | 0.35 | 4.8% | 0.54 | 7.5% | 0.00 | 0.0% | 0.76 | 10.5% | 1.00 | 13.8% | | | SCePr09 | 7.5 | 0.29 | 3.9% | 0.52 | 6.9% | 0.00 | 0.0% | 0.87 | 11.6% | 1.06 | 14.1% | | | SCePr02 | 14.6 | 0.54 | 3.7% | 1.31 | 9.0% | 0.00 | 0.0% | 1.43 | 9.8% | 2.01 | 13.8% | | | SCePr16 | 18.3 | 0.68 | 3.7% | 1.64 | 9.0% | 0.00 | 0.0% | 1.32 | 7.2% | 2.22 | 12.1% | | | SCePr19 | 39.5 | 2.28 | 5.8% | 2.72 | 6.9% | 0.73 | 1.9% | 5.66 | 14.3% | 6.72 | 17.0% | | | SCePr17 | 39.7 | 1.65 | 4.2% | 2.65 | 6.7% | 0.00 | 0.0% | 2.56 | 6.4% | 4.03 | 10.2% | | | SCePr08 | 86.0 | 3.97 | 4.6% | 5.60 | 6.5% | 0.00 | 0.0% | 10.55 | 12.3% | 12.58 | 14.6% | | | SCePr06 | 107.7 | 3.98 | 3.7% | 9.35 | 8.7% | 0.00 | 0.0% | 11.35 | 10.5% | 15.23 | 14.1% | | | SCePr07 | 196.6 | 9.12 | 4.6% | 15.30 | 7.8% | 0.00 | 0.0% | 19.19 | 9.8% | 26.18 | 13.3% | | | SCePr18 | 213.3 | 8.83 | 4.1% | 18.58 | 8.7% | 0.00 | 0.0% | 13.43 | 6.3% | 24.57 | 11.5% | | BioPlex® 2200 Celiac IgA – Anti-tTG IgA {11}------------------------------------------------ | | | | Within Run | | Between Run | | Between
Instrument/
Operator | | Between Lot | | Total | | |-----------|-------|------|------------|-------|-------------|------|------------------------------------|-------|-------------|-------|-------|--| | | Mean | | | | | | | | | | | | | Sample ID | U/mL | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | | SCePr09 | 1.5 | 0.10 | 6.3% | 0.16 | 10.3% | 0.00 | 0.0% | 0.00 | 0.0% | 0.19 | 12.1% | | | SCePr16 | 4.6 | 0.22 | 4.8% | 0.42 | 9.2% | 0.08 | 1.6% | 0.05 | 1.0% | 0.48 | 10.6% | | | SCePr14 | 15.4 | 0.67 | 4.3% | 1.03 | 6.6% | 0.00 | 0.0% | 0.52 | 3.4% | 1.33 | 8.6% | | | SCePr07 | 16.3 | 0.60 | 3.7% | 0.76 | 4.6% | 0.00 | 0.0% | 0.44 | 2.7% | 1.06 | 6.5% | | | SCePr03 | 33.3 | 1.22 | 3.7% | 1.54 | 4.6% | 0.00 | 0.0% | 1.60 | 4.8% | 2.54 | 7.6% | | | SCePr01 | 35.7 | 1.71 | 4.8% | 2.00 | 5.6% | 0.00 | 0.0% | 1.78 | 5.0% | 3.18 | 8.9% | | | SCePr22 | 99.3 | 4.38 | 4.4% | 5.87 | 5.9% | 0.00 | 0.0% | 6.67 | 6.7% | 9.90 | 10.0% | | | SCePr06 | 102.0 | 3.44 | 3.4% | 5.05 | 4.9% | 0.00 | 0.0% | 9.16 | 9.0% | 11.01 | 10.8% | | | SCePr05 | 188.9 | 6.86 | 3.6% | 11.88 | 6.3% | 0.00 | 0.0% | 15.81 | 8.4% | 20.93 | 11.1% | | | SCePr18 | 207.0 | 8.11 | 3.9% | 13.87 | 6.7% | 0.00 | 0.0% | 12.66 | 6.1% | 20.46 | 9.9% | | # BioPlex® 2200 Celiac IgA - Anti-DGP IgA # BioPlex® 2200 Celiac IgG – Anti-tTG IgG | | | | Within Run | | Between Run | | Between
Instrument/
Operator | | Between Lot | | Total | | |-----------|--------------|-------|------------|-------|-------------|------|------------------------------------|-------|-------------|-------|-------|------| | Sample ID | Mean
U/mL | SD | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | SCePr14 | 3.9 | 0.25 | 0.58 | 15.0% | 0.00 | 0.0% | 0.78 | 20.1% | 1.00 | 25.9% | 0.25 | 6.5% | | SCePr20 | 8.8 | 0.57 | 1.11 | 12.5% | 0.11 | 1.2% | 0.00 | 0.0% | 1.25 | 14.2% | 0.57 | 6.5% | | SCePr16 | 15.9 | 0.85 | 0.97 | 6.1% | 0.00 | 0.0% | 0.61 | 3.9% | 1.43 | 9.0% | 0.85 | 5.4% | | SCePr10 | 17.6 | 1.00 | 2.33 | 13.3% | 0.00 | 0.0% | 0.00 | 0.0% | 2.54 | 14.5% | 1.00 | 5.7% | | SCePr01 | 34.2 | 1.71 | 1.78 | 5.2% | 0.83 | 2.4% | 0.00 | 0.0% | 2.60 | 7.6% | 1.71 | 5.0% | | SCePr09 | 34.7 | 1.76 | 1.89 | 5.4% | 0.00 | 0.0% | 3.11 | 9.0% | 4.04 | 11.7% | 1.76 | 5.1% | | SCePr13 | 76.2 | 4.09 | 5.90 | 7.7% | 0.00 | 0.0% | 6.43 | 8.4% | 9.64 | 12.6% | 4.09 | 5.4% | | SCePr15 | 120.6 | 7.17 | 8.99 | 7.5% | 0.00 | 0.0% | 14.11 | 11.7% | 18.20 | 15.1% | 7.17 | 5.9% | | SCePr06 | 178.2 | 11.02 | 19.07 | 10.7% | 0.00 | 0.0% | 0.00 | 0.0% | 22.02 | 12.4% | 11.02 | 6.2% | | SCePr04 | 180.0 | 14.98 | 10.88 | 6.0% | 0.00 | 0.0% | 27.05 | 15.0% | 32.78 | 18.2% | 14.98 | 8.3% | ・ # BioPlex® 2200 Celiac IgG - Anti-DGP IgG · | BioPlex® 2200 Celiac IgG - Anti-DGP IgG | | | | | | | | | | | | | |-----------------------------------------|--------------|------------|------|-------------|-------|------------------------------------|------|-------------|-------|-------|-------|--| | | | Within Run | | Between Run | | Between
Instrument/
Operator | | Between Lot | | Total | | | | Sample ID | Mean
U/mL | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | | SCePr06 | 2.2 | 0.16 | 7.1% | 0.37 | 16.8% | 0.00 | 0.0% | 0.00 | 0.0% | 0.40 | 18.2% | | | SCePr07 | 10.2 | 0.59 | 5.7% | 0.56 | 5.5% | 0.20 | 2.0% | 0.42 | 4.1% | 0.94 | 9.2% | | | SCePr15 | 14.8 | 0.70 | 4.7% | 0.72 | 4.9% | 0.00 | 0.0% | 0.36 | 2.4% | 1.06 | 7.2% | | | SCePr10 | 15.5 | 0.83 | 5.3% | 0.92 | 6.0% | 0.00 | 0.0% | 0.92 | 5.9% | 1.54 | 9.9% | | | SCePr04 | 31.6 | 2.27 | 7.2% | 1.87 | 5.9% | 0.00 | 0.0% | 0.00 | 0.0% | 2.94 | 9.3% | | | SCePr14 | 32.6 | 1.45 | 4.5% | 1.27 | 3.9% | 0. ਰੋਰੇ | 3.0% | 1.98 | 6.1% | 1.27 | 3.9% | | | SCePr23 | 87.8 | 6.03 | 6.9% | 7.48 | 8.5% | 1.50 | 1.7% | 0.92 | 1.0% | 9.77 | 11.1% | | | SCePr13 | 92.1 | 4.82 | 5.2% | 5.86 | 6.4% | 0.00 | 0.0% | 2.14 | 2.3% | 7.88 | 8.6% | | | SCePr01 | 165.1 | 9.96 | 6.0% | 8.50 | 5.1% | 0.00 | 0.0% | 18.97 | 11.5% | 23.05 | 14.0% | | Page 12 of 21 {12}------------------------------------------------ # b. Linearity/assay reportable range: Three low and high Celiac anti-fTG IgA or IgG and anti-DGP IgA or IgG positive patient serum samples were tested to demonstrate linearity. These samples were diluted with immunodepleted serum according to CLSI EP06-A. Each sample and dilution was evaluated in replicates of four using one Celiac IgA and IgG lot on one instrument. Linear and polynomial regression analysis of Celiac IgA and IgG recovery vs. sample dilution was performed to determine if the dilution curves exhibit statistically significant non-linear regression based on the CLSI guideline EP06-A. | Celiac Assays | Sample | Conc. (U/mL) | Slope | Intercept | r2 | Dilution range (U/mL) | |---------------|--------|--------------|--------|-----------|--------|-----------------------| | Anti-tTG IgA | 1 | 58.4 | 1.0001 | -0.0016 | 0.9996 | 0.3 - 58.4 | | | 2 | 68.7 | 1.0000 | 0.0011 | 0.9991 | 0.3 - 68.7 | | | 3 | 61.8 | 1.0000 | 0.0005 | 0.9989 | 0.3 - 61.8 | | | 4 | 263.0 | 1.0000 | 0.0016 | 0.9963 | 0.3 - 263.0 | | | 5 | 235.1 | 1.0000 | 0.0016 | 0.9997 | 0.3 - 235.1 | | | 6 | 254.1 | 1.0000 | -0.0037 | 0.9984 | 0.3 - 254.1 | | Anti-DGP IgA | 1 | 60.9 | 1.0000 | 0.0006 | 0.9992 | 0.0 - 60.9 | | | 2 | 55.0 | 1.0001 | -0.0016 | 0.9988 | 0.0 - 55.0 | | | 3 | 72.6 | 1.0000 | -0.0011 | 0.9969 | 0.0 - 72.6 | | | 4 | 223.1 | 1.0000 | 0.0000 | 0.9997 | 0.0 - 223.1 | | | 5 | 206.2 | 1.0000 | 0.0015 | 0.9987 | 0.0 - 206.2 | | | 6 | 242.6 | 1.0000 | 0.0010 | 0.9995 | 0.0 - 242.6 | | Anti-tTG IgG | 1 | 67.5 | 1.0000 | 0.0011 | 0.9999 | 0.5 - 67.5 | | | 2 | 68.0 | 1.0000 | -0.0006 | 0.9995 | 0.5 - 68.0 | | | 3 | 46.7 | 1.0000 | -0.0010 | 0.9977 | 0.5 - 46.7 | | | 4 | 259.9 | 1.0000 | -0.0050 | 0.9984 | 0.3 - 259.9 | | | 5 | 232.7 | 1.0000 | -0.0001 | 0.9987 | 0.3 - 232.7 | | | 6 | 253.2 | 1.0000 | -0.0017 | 0.9972 | 0.3 - 253.2 | | Anti-DGP IgG | 1 | 48.5 | 1.0000 | 0.0016 | 0.9997 | 0.0 - 48.5 | | | 2 | 57.9 | 1.0000 | -0.0005 | 0.9989 | 0.0 - 57.9 | | | 3 | 49.2 | 1.0000 | 0.0000 | 0.9982 | 0.0 - 49.2 | | | 4 | 192.9 | 1.0000 | -0.0017 | 0.9985 | 0.0 - 192.9 | | | 5 | 183.4 | 1.0000 | 0.0000 | 0.9992 | 0.0 - 183.4 | | | 6 | 238.6 | 1.0000 | 0.0017 | 0.9984 | 0.0 - 238.6 | The regression parameters (slope, intercept and r2) of the observed values vs. nredicted values are show below | BioPlex 2200 Celiac Assay | Assay Measuring Range | |---------------------------|-----------------------| | Anti-tTG IgA | 0.5 to 250 U/mL | | Anti-DGP IgA | 0.2 to 250 U/mL | | Anti-tTG IgG | 0.8 to 250 U/mL | | Anti-DGP IgG | 0.4 to 250 U/mL | Traceability, Stability, Expected values (controls, calibrators, or methods): C. {13}------------------------------------------------ ### Traceability: There is no international or certified reference material available for Celiac antitTG and anti-DGP IgA and IgG. The calibrators are assigned relative arbitrary units (U/mL). ### Value Assignment: The calibrators are manufactured independently from the controls, and are stabilized with <0.3% ProClin® 300, <0.1% sodium benzoate, and <0.1% sodium azide. Calibrator assignment is established for matched lots of BioPlex® 2200 Celiac IgA or IgG kit and calibrators using a master set of calibrators as reference and replicate analyses on multiple BioPlex® 2200 instruments. The BioPlex® 2200 Celiac IgA or IgG Reagent Kit is calibrated using a set of five (5) distinct calibrators for anti-tTG and anti-DGP IgG or IgA. which are used to establish points of reference for determining the presence of anti-tTG and anti-DGP IgA or IgG in human specimens. The negative control is manufactured to give negative results with values below the cut-off for each assay. The positive control is manufactured to give positive results with values above the assay cut-off for each assay. ### Stability: Stability studies have been performed to support the following claims: ### Calibrator and Control: BioPlex® 2200 Celiac IgA and IgG Control and Calibrator Sets: Calibrator Open Vial Stability (2 to 8°C). 60 days from first opening: Control Open Vial Stability (2 to 8°C), 60 days from first opening; Calibration Curve On-board Stability, 30 days; Calibrators and Controls Real Time Stability (2 to 8°C), 9 months; labeled as until expiration date. ### Kit Stability: BioPlex® 2200 Celiac IgA and IgG Kit: Real Time (unopened) Kit Stability, 9 months or until the date of expiration when stored unopened on the instrument or at 2 to 8°C; The Open kit claim is 60 days. ### Serum Sample Stability: Sample stability studies were also performed: Sample stability fresh (2 to 8°C), 7 days; Sample stability frozen (-20 or -80°C), 8 months; Sample Freeze-thaw (-20 or -80℃), up to 3-freeze thaw cycles acceptable. ### d. Detection limit: The results of LoQ, LoD and LoB are summarized in the table below. | BioPlex® 2200 Celiac
Assay | LoQ
(U/mL) | LoD
(U/mL) | LoB
(U/mL) | |-------------------------------|---------------|---------------|---------------| | Anti-tTG IgA | 0.5 | 0.5 | 0.4 | | Anti-DGP IgA | 0.2 | 0.1 | 0.0 | {14}------------------------------------------------ | BioPlex® 2200 Celiac
Assay | LoQ
(U/mL) | LoD
(U/mL) | LoB
(U/mL) | |-------------------------------|---------------|---------------|---------------| | Anti-tTG IgG | 0.8 | 0.8 | 0.6 | | Anti-DGP IgG | 0.4 | 0.1 | 0.0 | e. Analytical specificity: An interfering substances study was conducted to evaluate the potential interference of specific endogenous and exogenous substances with the BioPlex® 2200 Celiac IgA and IgG kits according to CLSI EP7-A2. No interference was observed with any of the substances tested, except anti-tTG IgA and IgG at hemolysate level above hemoglobin concentration of 63 mg/dL. The substances and the maximum levels tested are shown in the table below: | Substance | Concentration | |--------------------------|---------------| | Hemolysate | <500 mg/dL* | | Bilirubin (unconjugated) | <20 mg/dL | | Bilirubin (conjugated) | <30 mg/dL | | Triglycerides | <3300 mg/dL | | Total Protein | <12 g/dL | | Cholesterol | <500 mg/dL | | Red Blood Cells | <0.4% (v/v) | | Gamma-Globulin | << g/dL | | Beta-Carotene | <0.6 mg/dL | | Ascorbic Acid | <3 mg/dL | | EDTA | < 800 mg/dL | *for anti-DGP IgA and IgG only ### Cross-Reactivity: A cross-reactivity study was performed to determine if patient serum samples from individuals with various disease states and other potentially interfering factors interfere with test results from the BioPlex® 2200 Celiac IgA or IgG kit. Samples from individuals with known disease states for potential cross reactivity listed in the table below were evaluated with the BioPlex® 2200 Celiac IgA or IgG kit. The table below shows the number (N) of samples containing potential cross reactants as disease state evaluated by the BioPlex® Celiac IgA and IgG kits. The cross reactivity was obtained as the positivity rate from the ratio of the number of samples scored positive by the BioPlex® Celiac IgA and IgG assays to the total number of cross reactant samples evaluated. {15}------------------------------------------------ | | N | BioPlex 2200 | | | | | | | | |---------------------------------|-----|--------------|-----------------|--------------|-----------------|--------------|-----------------|--------------|-----------------| | Cross Reactive
Disease State | | Anti-tTG IgA | | Anti-DGP IgA | | Anti-tTG IgG | | Anti-DGP IgG | | | | | Pos (+) | %
Positivity | Pos (+) | %
Positivity | Pos (+) | %
Positivity | Pos (+) | %
Positivity | | Chronic Active Hepatitis | 20 | 0 | 0% | 0 | 0% | 5 | 25% | 0 | 0% | | Crohn's Disease | 23 | 1 | 4% | 1 | 4% | 0 | 0% | 0 | 0% | | Diabetes Mellitus Type 1 | 20 | 0 | 0% | 0 | 0% | 0 | 0% | 0 | 0% | | Gastritis | 20 | 0 | 0% | 0 | 0% | 0 | 0% | 0 | 0% | | Graves'/Hashimoto's Disease | 23 | 0 | 0% | 1 | 4% | 0 | 0% | 1 | 4% | | Irritable Bowel Syndrome | 15 | 0 | 0% | 0 | 0% | 0 | 0% | 0 | 0% | | Pernicious Anemia | 6 | 0 | 0% | 0 | 0% | 0 | 0% | 0 | 0% | | Primary Biliary Cirrhosis | 19 | 0 | 0% | 0 | 0% | 0 | 0% | 2 | 11% | | Rheumatoid Arthritis | 20 | 1 | 5% | 2 | 10% | 1 | 5% | 1 | 5% | | Scleroderma | 21 | 0 | 0% | 1 | 5% | 0 | 0% | 1 | 5% | | Sjögren's Syndrome | 24 | 0 | 0% | 1 | 4% | 1 | 4% | 1 | 4% | | Syphilis | 10 | 0 | 0% | 1 | 10% | 0 | 0% | 0 | 0% | | Systemic Lupus Erythematosus | 30* | 0 | 0% | 0 | 0% | 0 | 0% | 0 | 0% | | Ulcerative Colitis | 22 | 0 | 0% | 0 | 0% | 0 | 0% | 0 | 0% | * 1 sample exhibited repeated instrument error flags and was excluded from the anti-tTG IgA and IgG data analysis #### f. Assay cut-off: The assay cutoff values were determined by performing Receiver Operator Characteristic (ROC) analysis using clinically diagnosed serum samples. The study to determine the Celiac IgA or IgG assay cutoff was comprised of two patient serum sample groups -123 samples from patients diagnosed with celiac disease and 112 from non-celiac or other rheumatic disease control donors. The assay cut-off values were confirmed by testing 298 samples from apparently healthy donors. A cutoff of 15.0 U/mL for anti-tTG IgA or IgG and for anti-DGP IgA or IgG was established by optimizing for clinical accuracy. #### 2. Comparison studies: ### a. Method comparison with predicate device: The performance of the BioPlex® 2200 Celiac IgA and IgG kits was evaluated including 156 patients diagnosed with celiac disease 163 patients with other Page 16 of 21 {16}------------------------------------------------ rheumatic or non-CD disease control, 11 celiac IgA deficient patients, and 16 dermatitis herpetiformis (DH) patients. Results from all patient serum samples of both the new and the predicate immunoassays are compared. Results are summarized in the tables below: ### All Retrospective Patient Serum Samples BioPlex 2200 Celiac IgA BioPlex 2200 Celiac IgG | | | Predicate IgA EIA Kit | | | |--------------|----------|-----------------------|----------|-------| | | | Positive | Negative | Total | | anti-tTG IgA | Positive | 163 | 0 | 163 | | | Negative | 6 | 176 | 182 | | | Total | 169 | 176 | 345* | | anti-DGP IgA | Positive | 144 | 13 | 157 | | | Negative | 4 | 185 | 189 | | | Total | 148 | 198 | 346 | * 1 sample exhibited repeated instrument error flags and was excluded from the data analysis · Anti-tTG IgA Positive Agreement (95% CI) = 96.4% (163/169) (92.5 - 98.4%) Anti-tTG IgA Negative Agreement (95% Cl) = 100% (176/176) (97.9 - 100%) Anti-tTG IgA Total Agreement (95% CI) = 98.3% (339/345) (96.3 - 99.2%) Anti-DGP IgA Positive Agreement (95% CI) = 97.3% (144/148) (93.3 - 98.9%) Anti-DGP IgA Negative Agreement (95% CI) = 93.4% (185/198) (89.1 - 96.1%) Anti-DGP IgA Total Agreement (95% CI) = 95.1% (329/346) (92.3 - 96.9%) | | | Predicate IgG EIA Kit | | | |--------------|----------|-----------------------|----------|-------| | | | Positive | Negative | Total | | anti-tTG IgG | Positive | 56 | 36 | 92 | | | Negative | 4 | 249 | 253 | | | Total | 60 | 285 | 345* | | anti-DGP IgG | Positive | 140 | 17 | 157 | | | Negative | 10 | 179 | 189 | | | Total | 150 | 196 | 346 | * 1 sample exhibited repeated instrument error flags and was excluded from the data analysis Anti-tTG IgG Positive Agreement (95% CI) = 93.3% (56/60) (84.1-97.4%) Anti-tTG IgG Negative Agreement (95% C1) = 87.4% (249/285) (83.0 - 90.7%) Anti-tTG IgG Total Agreement (95% CI) = 88.4% (305/345) (84.6-91.4%) Anti-DGP IgG Positive Agreement (95% CI) = 93.3% (140/150) (88.2 - 96.3%) Page 17 of 21 {17}------------------------------------------------ Anti-DGP IgG Negative Agreement (95% Cl) = 91.3% (179/196) (86.6 - 94.5%) Anti-DGP IgG Total Agreement (95% CI) = 92.2% (319/346) (88.9 - 94.6%) Results from values in the measuring range and 10% of diluted total samples of both the new and the predicate immunoassays are compared. Results are summarized in the tables below: # Results within the measuring range and 10% of diluted serum samples BioPlex 2200 Celiac IgA | | | Predicate IgA EIA Kit | | | |----------------------------------|----------|-----------------------|----------|-------| | | | Positive | Negative | Total | | anti-tTG IgA
(0.5 - 250 U/mL) | Positive | 97 | 0 | 97 | | | Negative | 6 | 108 | 114 | | | Total | 103 | 108 | 211 | | anti-DGP IgA
(0.2 - 250 U/mL) | Positive | 129 | 13 | 142 | | | Negative | 4 | 164 | 168 | | | Total | 133 | 177 | 310 | Anti-tTG IgA Positive Agreement (95% CI) = 94.2% (97/103) (87.9 - 97.3%) Anti-tTG IgA Negative Agreement (95% Cl) = 100% (108/108) (96.6 - 100%) Anti-tTG IgA Total Agreement (95% CI) = 97.2% (205/211) (93.9 - 98.7%) Anti-DGP IgA Positive Agreement (95% CI) = 97.0% (129/133) (92.5 - 98.8%) Anti-DGP IgA Negative Agreement (95% Cl) = 92.7% (164/177) (87.8 - 95.7%) Anti-DGP IgA Total Agreement (95% CI) = 94.5% (293/310) (91.4 - 96.5%) BioPlex 2200 Celiac IgG | | | Predicate IgG EIA Kit | | | |----------------------------------|----------|-----------------------|----------|-------| | | | Positive | Negative | Total | | anti-tTG IgG
(0.8 - 250 U/mL) | Positive | 54 | 36 | 90 | | | Negative | 4 | 194 | 198 | | | Total | 58 | 230 | 288 | | anti-DGP IgG
(0.4 - 250 U/mL) | Positive | 140 | 17 | 157 | | | Negative | 9 | 82 | 91 | | | Total | 149 | 99 | 248 | Anti-tTG IgG Positive Agreement (95% CI) = 93.1% (54/58) (83.6 - 97.3%) Anti-tTG IgG Negative Agreement (95% Cl) = 84.3% (194/230) (79.1 - 88.5%) Anti-tTG IgG Total Agreement (95% CI) = 86.1% (248/288) (81.6 - 89.6%) Anti-DGP IgG Positive Agreement (95% Cl) = 94.0% (140/149) (88.9 - 96.8%) Anti-DGP IgG Negative Agreement (95% CI) = 82.8% (82/99) (74.2 -- 89.0%) Anti-DGP IgG Total Agreement (95% CI) = 89.5% (222/248) (85.1 - 92.7%) Page 18 of 21 {18}------------------------------------------------ - b. Matrix comparison: Not Applicable- Serum only. #### 3. Clinical studies: - a. Clinical Sensitivity and specificity: The clinical studies involved testing 319 serum specimens including 163 non-Celiac disease control patients and 156 diagnosed Celiac disease patients. The BioPlex® 2200 Celiac IgA and IgG sensitivity and specificity are shown below: | | | Clinical Diagnosis | | | |--------------|----------|--------------------|----------|-------| | | | Positive | Negative | Total | | anti-tTG IgA | Positive | 148 | 2 | 150 | | | Negative | 8 | 160 | 168 | | | Total | 156 | 162 | 318* | | anti-DGP IgA | Positive | 136 | 5 | 141 | | | Negative | 20 | 158 | 178 | | | Total | 156 | 163 | .319 | BioPlex 2200 Celiac IgA * 1 sample exhibited repeated instrument error flags and was excluded from the data analysis Anti-tTG IgA Sensitivity (95% Cl) = 94.9% (148/156) (90.2 - 97.4%) Anti-tTG IgA Specificity (95% CI) = 98.8% (160/162) (95.6 - 99.7%) Anti-DGP IgA Sensitivity (95% CI) = 87.2% (136/156) (81.0 - 91.5%) Anti-DGP IgA Specificity (95% CI) = 96.9% (158/163) (93.0 - 98.7%) | | | Clinical Diagnosis | | | |--------------|----------|--------------------|----------|-------| | | | Positive | Negative | Total | | anti-tTG IgG | Positive | 69 | 7 | 76 | | | Negative | 87 | 155 | 242 | | | Total | 156 | 162 | 318* | | anti-DGP IgG | Positive | 132 | 5 | 137 | | | Negative | 24 | 158 | 182 | | | Total | 156 | 163 | 319 | BioPlex 2200 Celiac IgG * 1 sample exhibited repeated instrument error flags and was excluded from the data analysis Anti-tTG IgG Sensitivity (95% CI) = 44.2% (69/156) (36.7 - 52.1%) Page 19 of 21 {19}------------------------------------------------ Anti-tTG IgG Specificity (95% CI) = 95.7% (155/162) (91.4 - 97.9%) Anti-DGP IgG Sensitivity (95% CI) = 84.6% (132/156) (78.1 - 89.4%) Anti-DGP IgG Specificity (95% CI) = 96.9% (158/163) (93.0 - 98.7%) Serum samples from patients previously diagnosed with celiac disease and IgA deficiency (N=11) were run on the BioPlex 2200 Celiac IgG kits and the # positive samples is shown in the table below: | Patient Group | # Positive Anti-tTG IgG (%) | # Positive Anti-DGP IgG (%) | |---------------------------------------|-----------------------------------|-----------------------------------| | Celiac Disease with
IgA deficiency | 9 (81.8%)
95% CI: 52.3 – 94.9% | 8 (72.7%)
95% CI: 43.4 – 90.3% | The positive rates of the BioPlex Celiac IgA and IgG in each of disease category are shown below. | Disease Category | Number Enrolled | Anti-tTG IgA | Anti-DGP IgA | Anti-tTG IgG | Anti-DGP IgG | |--------------------------------|-----------------|----------------|----------------|---------------|----------------| | Celiac Disease | 156 | 148
(94.9%) | 136
(87.2%) | 69
(44.2%) | 132
(84.6%) | | Apparently Healthy Subject | 300 | 0
(0.0%) | 5
(1.7%) | 2
(0.7%) | 5
(1.7%) | | IgA Deficiency | 11 | 0
(0.0%) | 0
(0.0%) | 9
(81.8%) | 8
(72.7%) | | Dermatitis Herpetiformis | 16 | 13
(81.3%) | 16
(100%) | 7
(43.8%) | 12
(75.0%) | | Chronic Active Hepatitis | 10 | 0
(0%) | 0
(0%) | 5 (50%) | 0
(0%) | | Crohn's Disease | 13 | 1
(7.7%) | 0
(0%) | 0
(0%) | 0
(0%) | | Diabetes Mellitus Type 1 | 10 | 0
(0%) | 0
(0%) | 0
(0%) | 0
(0%) | | Gastritis | 10 | 0
(0%) | 0
(0%) | 0
(0%) | 0
(0%) | | Graves'/Hashimoto's
Disease | 13 | 0
(0%) | 1
(7.7%) | 0
(0%) | 0
(0%) | | Irritable Bowel Syndrome | 15 | 0
(0%) | 0
(0%) | 0
(0%) | 0
(0%) | | Pernicious Anemia | 6 | 0
(0%) | 0
(0%) | 0
(0%) | 0
(0%) | | Primary Biliary Cirrhosis | 9 | 0
(0%) | 0
(0%) | 0
(0%) | 2
(22.2%) | | Rheumatoid Arthritis | 10 | 1
(10%) | 1
(10%) | 1
(10%) | 1
(10%) | | Scleroderma | 11 | 0
(0%) | 1
(9.1%) | 0
(0%) | 1
(9.1%) | | Sjögren's Syndrome | 14 | 0
(0%) | 1
(7.1%) | 1
(7.1%) | 1
(7.1%) | | Systemic Lupus Erythematosus | 20* | 0
(0%) | 0
(0%) | 0
(0%) | 0
(0%) | | Ulcerative Colitis | 12 | 0
(0%) | 0
(0%) | 0
(0%) | 0
(0%) | Page 20 of 21 {20}------------------------------------------------ | Disease Category | Number | Anti-tTG | Anti-DGP Anti-tTG Anti-DGP | | | |------------------|----------|----------|--------------------------------|------|------| | | Enrolled | IgA · | IgA | lgG | IgG | | Syphilis | 10 | | | | | | | | (0%) | (10%) | (0%) | (0%) | * 1 sample exhibited repeated instrument error flags and was excluded from the anti-tTG IgG and IgA data analysis - c. Other clinical supportive data (when a. and b. are not applicable): Refer to Method Comparison - 4. Clinical cut-off: See Assay Cutoff - 5. Expected values/Reference range: Three hundred serum samples from apparently healthy donors including 139 males ranging in age from <1 to 94 and 161 females ranging in age from 5 to 101 were tested with BioPlex® 2200 Celiac IgA and IgG kits. The number of positive and mean value of the BioPlex® Celiac IgA and IgG results are shown below. | Assay | N (%Positive) | Mean (U/mL) | |--------------|---------------|-------------| | anti-tTG IgA | 0 (0.0%) | 0.6 | | anti-DGP IgA | 5 (1.7%) | 2.2 | | anti-tTG IgG | 2 (0.7%) | 2.3 | | anti-DGP IgG | 5 (1.7%) | 1.7 | Results of <15.0 U/mL for anti-tTG IgA or IgG and for anti-DGP IgA or IgG are reported as negative and results >15.0 U/mL for anti-tTG IgA or IgG and for anti-DGP IgA or IgG are reported as positive. Each laboratory should establish its own reference range pertinent to their specific patient populations. ### Instrument Name: The BioPlex 2200 System, software version 4.1 {21}------------------------------------------------ ### DEPARTMENT OF HEALTH & HUMAN SERVICES Image /page/21/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized eagle with three wavy lines extending from its body, representing the department's commitment to health and well-being. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular pattern around the eagle symbol. The logo is simple, clean, and easily recognizable. Public Health Service Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002 September 19, 2013 BIO-RAD LABORATORIES, INC. C/O DR. JUANG WANG REGULATORY AFFAIRS REPRESENTATIVE 5500 EAST 2ND ST. BENICIA, CA 94510 Re: K130053 Trade/Device Name: BioPlex® 2200 Celiac IgA kit BioPlex® 2200 Celiac lgG kit BioPlex® 2200 Celiac IgA Calibrator Set BioPlex® 2200 Celiac IgG Calibrator Set BioPlex® 2200 Celiac IgA Control Set BioPlex® 2200 Celiac IgG Control Set Regulation Number: 21 CFR 866.5660 Regulation Name: Multiple Autoantibodies Immunological Test System Regulatory Class: II Product Code: MVM, MST, JIX, JJX Dated: August 22, 2013 Received: August 23, 2013 Dear Dr. Wang: We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. 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 additiona… --- **Source:** [https://fda.innolitics.com/submissions/IM/subpart-f%E2%80%94immunological-test-systems/MVM/K130053](https://fda.innolitics.com/submissions/IM/subpart-f%E2%80%94immunological-test-systems/MVM/K130053) **Published by [Innolitics](https://innolitics.com)** — a medical-device software consultancy. We help companies design, build, and clear FDA-regulated software and AI/ML devices. If you're preparing [a 510(k)](https://innolitics.com/services/510ks/), [a De Novo](https://innolitics.com/services/regulatory/), [a SaMD](https://innolitics.com/services/end-to-end-samd/), [an AI/ML medical device](https://innolitics.com/services/medical-imaging-ai-development/), or [an FDA regulatory strategy](https://innolitics.com/services/regulatory/), [get in touch](https://innolitics.com/contact). **Cite:** Innolitics at https://innolitics.com