(247 days)
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.
The BioPlex 2200 Celiac IgA Calibrator Set is intended for the calibration of the BioPlex 2200 Celiac IgA Reagent Pack.
The BioPlex 2200 Celiac IgG Calibrator Set is intended for the calibration of the BioPlex 2200 Celiac IgG Reagent Pack.
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.
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.
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.
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.
Here's an analysis of the acceptance criteria and study findings for the BioPlex® 2200 Celiac IgA and IgG kits, based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance:
The document doesn't explicitly state numerical acceptance criteria in a dedicated section with pass/fail thresholds. Instead, it presents performance characteristics from various studies. For this table, I will present the reported performance, which implicitly serves as the evidence that the device meets acceptable levels for these metrics. The "Target Performance" column will reflect typical expectations for such assays or the performance shown by the predicate devices where applicable.
Performance Characteristics for BioPlex® 2200 Celiac IgA and IgG Kits
| Performance Metric | Specific Assay | Reported Device Performance (Mean %CV/Values) | Target Performance (Implicit/Predicate) |
|---|---|---|---|
| Precision/Reproducibility (CLSI EP5-A2, 20 days, n=80 per sample type) | Anti-tTG IgA | Total Precision: 7.5% - 10.9% CV | Within generally accepted ranges for immunoassays (typically <15-20% CV for quantitative assays, with controls often <10% CV) |
| Anti-DGP IgA | Total Precision: 5.4% - 11.0% CV | ||
| Anti-tTG IgG | Total Precision: 5.0% - 15.4% CV | ||
| Anti-DGP IgG | Total Precision: 4.2% - 11.2% CV | ||
| Precision/Reproducibility (CLSI EP15-A2, 5 days, n=20 per sample type) | Anti-tTG IgA | Total Precision: 4.4% - 7.5% CV | Similar to above |
| Anti-DGP IgA | Total Precision: 3.6% - 6.3% CV | ||
| Anti-tTG IgG | Total Precision: 3.0% - 9.8% CV | ||
| Anti-DGP IgG | Total Precision: 2.8% - 8.6% CV | ||
| Lot-to-lot Reproducibility (3 lots, 3 inst., 2 runs, 10 replicates, n=60) | Anti-tTG IgA | Total Precision: 10.2% - 17.0% CV | Typically <20% CV for lot-to-lot variability for quantitative assays |
| Anti-DGP IgA | Total Precision: 6.5% - 12.1% CV | ||
| Anti-tTG IgG | Total Precision: 8.3% - 25.9% CV | ||
| Anti-DGP IgG | Total Precision: 7.2% - 18.2% CV | ||
| Linearity/Reportable Range | Anti-tTG IgA | 0.5 to 250 U/mL (slopes ~1.000, r² ~0.99) | Demonstrated linearity across the claimed measuring range. |
| Anti-DGP IgA | 0.2 to 250 U/mL (slopes ~1.000, r² ~0.99) | ||
| Anti-tTG IgG | 0.8 to 250 U/mL (slopes ~1.000, r² ~0.99) | ||
| Anti-DGP IgG | 0.4 to 250 U/mL (slopes ~1.000, r² ~0.99) | ||
| Detection Limit | Anti-tTG IgA | LoQ: 0.5 U/mL, LoD: 0.5 U/mL, LoB: 0.4 U/mL | Typically, LoQ > LoD > LoB, and values should be analytically sound for the intended use. |
| Anti-DGP IgA | LoQ: 0.2 U/mL, LoD: 0.1 U/mL, LoB: 0.0 U/mL | ||
| Anti-tTG IgG | LoQ: 0.8 U/mL, LoD: 0.8 U/mL, LoB: 0.6 U/mL | ||
| Anti-DGP IgG | LoQ: 0.4 U/mL, LoD: 0.1 U/mL, LoB: 0.0 U/mL | ||
| Clinical Sensitivity | Anti-tTG IgA | 94.9% (148/156) (95% CI: 90.2 - 97.4%) | High sensitivity for aid in diagnosis of Celiac Disease. |
| Anti-DGP IgA | 87.2% (136/156) (95% CI: 81.0 - 91.5%) | ||
| Anti-tTG IgG | 44.2% (69/156) (95% CI: 36.7 - 52.1%) | ||
| Anti-DGP IgG | 84.6% (132/156) (95% CI: 78.1 - 89.4%) | ||
| Clinical Specificity | Anti-tTG IgA | 98.8% (160/162) (95% CI: 95.6 - 99.7%) | High specificity for aid in diagnosis of Celiac Disease. |
| Anti-DGP IgA | 96.9% (158/163) (95% CI: 93.0 - 98.7%) | ||
| Anti-tTG IgG | 95.7% (155/162) (95% CI: 91.4 - 97.9%) | ||
| Anti-DGP IgG | 96.9% (158/163) (95% CI: 93.0 - 98.7%) | ||
| Method Comparison (Predicate Device) | Anti-tTG IgA (Positive Agreement) | 96.4% (163/169) (95% CI: 92.5 - 98.4%) | High agreement with legally marketed predicate devices. |
| Anti-tTG IgA (Negative Agreement) | 100% (176/176) (95% CI: 97.9 - 100%) | ||
| Anti-tTG IgA (Total Agreement) | 98.3% (339/345) (95% CI: 96.3 - 99.2%) | ||
| Anti-DGP IgA (Positive Agreement) | 97.3% (144/148) (95% CI: 93.3 - 98.9%) | ||
| Anti-DGP IgA (Negative Agreement) | 93.4% (185/198) (95% CI: 89.1 - 96.1%) | ||
| Anti-DGP IgA (Total Agreement) | 95.1% (329/346) (95% CI: 92.3 - 96.9%) | ||
| Anti-tTG IgG (Positive Agreement) | 93.3% (56/60) (95% CI: 84.1-97.4%) | ||
| Anti-tTG IgG (Negative Agreement) | 87.4% (249/285) (95% CI: 83.0 - 90.7%) | ||
| Anti-tTG IgG (Total Agreement) | 88.4% (305/345) (95% CI: 84.6-91.4%) | ||
| Anti-DGP IgG (Positive Agreement) | 93.3% (140/150) (95% CI: 88.2 - 96.3%) | ||
| Anti-DGP IgG (Negative Agreement) | 91.3% (179/196) (95% CI: 86.6 - 94.5%) | ||
| Anti-DGP IgG (Total Agreement) | 92.2% (319/346) (95% CI: 88.9 - 94.6%) |
2. Sample Size Used for the Test Set and Data Provenance:
-
Precision/Reproducibility:
- CLSI EP5-A2: 24 samples (serum panel consisting of samples spanning the measuring range), tested in replicate twice daily over 20 days (n=80 replicates per sample type).
- CLSI EP15-A2: 12 samples (serum panel consisting of samples spanning the measuring range), tested in 4 replicates per run, one run per day over 5 days (n=20 replicates per sample type).
- Lot-to-lot Reproducibility: Unspecified number of serum samples covering the assay range, tested with three reagent lots on three instruments, 10 replicates for two runs per lot (60 points per sample ID for calculation).
-
Linearity/Reportable Range: 3 low positive and 3 high positive patient serum samples for each antibody (anti-tTG IgA/IgG, anti-DGP IgA/IgG).
-
Analytical Specificity (Interference): Test substances (e.g., Hemolysate, Bilirubin, Triglycerides). No specific number of samples given for the interference study, but the substances were tested at specified concentrations.
-
Analytical Specificity (Cross-Reactivity): 244 samples from individuals with various disease states (e.g., Chronic Active Hepatitis, Crohn's Disease, Diabetes Mellitus Type 1, Rheumatoid Arthritis). The number of samples for each disease state ranged from 6 to 30.
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Assay Cut-off Determination: 123 samples from patients with diagnosed celiac disease and 112 from non-celiac/rheumatic disease control donors. Confirmed with 298 samples from apparently healthy donors.
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Method Comparison with Predicate Device:
- All samples: 156 patients with celiac disease, 163 patients with other rheumatic or non-CD disease control, 11 celiac IgA deficient patients, and 16 Dermatitis Herpetiformis (DH) patients. Total: 346-347 samples (1 sample excluded in some analyses).
- Samples within measuring range (and 10% diluted): Varying totals per assay, e.g., anti-tTG IgA (211), anti-DGP IgA (310), anti-tTG IgG (288), anti-DGP IgG (248).
-
Clinical Studies (Sensitivity and Specificity): 319 serum specimens, including 163 non-Celiac disease control patients and 156 diagnosed Celiac disease patients.
-
Expected Values/Reference Range: 300 serum samples from apparently healthy donors (139 males, 161 females, age <1 to 101).
Data Provenance (Retrospective/Prospective, Country of Origin):
The document states that the method comparison study used "all retrospective patient serum samples." Similarly, the clinical sensitivity and specificity study involved clinical diagnosis of Celiac patients and non-Celiac controls. For cross-reactivity and expected values, these also appear to be retrospective collections of patient/donor samples. There is no explicit mention of the country of origin for the data.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts:
The document does not explicitly state the number of experts or their specific qualifications (e.g., "radiologist with 10 years of experience") for establishing the ground truth.
For the "clinical diagnosis," it refers to "patients previously diagnosed with celiac disease" and "non-Celiac disease control patients." This implies that the diagnosis was established through standard clinical practice by relevant healthcare professionals (e.g., gastroenterologists) using a combination of clinical findings, biopsy, and other diagnostic tests, which is a common approach for establishing ground truth in diagnostic assay validation. The specific expertise used for these previous diagnoses is not detailed.
4. Adjudication Method for the Test Set:
The document does not describe a specific adjudication method like "2+1" or "3+1" for determining ground truth in the patient samples. The ground truth appears to be based on "clinical diagnosis" or categorization of "apparently healthy donors" and specific disease states. While this is a form of ground truth, it wasn't established through a formal adjudication process (e.g., blinded review by multiple experts with a tie-breaker).
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done, If So, What Was the Effect Size of How Much Human Readers Improve With AI vs Without AI Assistance:
This question is not applicable. The device is an in vitro diagnostic immunoassay for semi-quantitative detection of antibodies, not an imaging device or AI system that assists human "readers" in interpreting images. Therefore, an MRMC study or an assessment of human reader improvement with AI assistance is not relevant to this product.
6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done:
This question is not directly applicable in the typical sense of AI algorithms. The BioPlex® 2200 system performs the assay and provides numerical results (U/mL) without human interpretive input for the final measurement. The "standalone" performance is essentially the output of the instrument-reagent system. The "clinical sensitivity and specificity" study (Section 3) represents the standalone performance of the assay compared to clinical diagnosis.
7. The Type of Ground Truth Used (Expert Consensus, Pathology, Outcomes Data, etc.):
The ground truth used for performance evaluation can be categorized as:
- Clinical Diagnosis: For the clinical sensitivity and specificity studies, the ground truth for "Celiac disease patients" and "non-Celiac disease control patients" was based on their clinical diagnosis. This typically involves a combination of symptoms, serology, and duodenal biopsy.
- Predicate Device Results: For the method comparison study, the predicate immunoassay results served as the comparison point for agreement. While not a "true" ground truth in a clinical sense, it acts as a reference for establishing substantial equivalence.
- Disease State Classification: For cross-reactivity studies, samples were from individuals with "known disease states."
- Apparently Healthy Donors: For establishing expected values and confirming cut-offs, samples were from "apparently healthy donors."
8. The Sample Size for the Training Set:
The document does not explicitly describe a separate "training set" in the context of machine learning (AI). This is a diagnostic immunoassay system. The development of the assay (e.g., antigen selection, calibrator assignment, and establishment of linearity) inherently involves a "training" or development phase, but it's not described as a discrete "training set" in the AI sense.
However, the "Assay cut-off" section mentions that the cut-off values were determined using "123 samples from patients diagnosed with celiac disease and 112 from non-celiac or other rheumatic disease control donors." This set of samples was used to perform ROC analysis to establish the cut-off, which is a critical parameter for classifying results and could be considered analogous to a development or "training" set for defining the decision boundary of the assay. This was then "confirmed by testing 298 samples from apparently healthy donors."
9. How the Ground Truth for the Training Set Was Established:
As discussed in point 8, the "training set" for establishing the assay cut-off involved:
- Clinical Diagnosis: For the 123 celiac disease patients and 112 non-celiac/rheumatic disease control donors, the ground truth was their established clinical diagnosis. This implies diagnosis by medical professionals using standard diagnostic criteria.
- Healthy Status: For the 298 apparently healthy donors, the ground truth was their healthy status, likely determined through health screenings or self-reporting.
The specific "how" (e.g., detailed diagnostic reports, biopsy confirmation) and the expertise involved in these diagnoses are not explicitly detailed in the document beyond "clinically diagnosed."
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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
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Regulatory Information:
| Product Code | Classification | Regulation Section | Panel |
|---|---|---|---|
| Autoantibodies,endomysial(tissuetransglutaminase)(MVM) | Class II | 21 CFR § 866.5660,Multiple autoantibodiesimmunological test system. | Immunology (82) |
| Antibodies, Gliadin(MST) | Class II | 21 CFR § 866.5750,Radioallergosorbent(RAST) immunological testsystem. | Immunology (82) |
| Calibrator, Multi-AnalyteMixture(JIX) | Class II | 21 CFR § 862.1150 -Calibrator | Clinical Chemistry(75) |
| Multi-Analyte ControlsAll 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.
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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.
-
- Indication(s) for use: Same as Intended Use
-
- Special conditions for use statement(s): For prescription use only
-
- 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.
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- 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
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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 andIgG Kits | Predicate:INOVA QUANTA Liteh-tTG IgA and IgGKits | Predicate:INOVA QUANTA LiteGliadin IgA II and IgGII Kits | |
| Assay Technology | Automated multiplex flowimmunoassay | Manual, microtiterplate format,Enzyme-linkedImmunosorbent assay(ELISA) | Manual, microtiterplate format,Enzyme-linkedImmunosorbent assay(ELISA) |
| Capture Antigen | Recombinant tTG proteinand DGP antigen | Native human tissuetransglutaminase (h-tTG) | Synthetic,deamidated peptide |
| Conjugate | Phycoerythrin conjugated | Goat anti-human IgG | Goat anti-human IgG |
·
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| Differences | ||||||||
|---|---|---|---|---|---|---|---|---|
| ltem | Device:BioPlex 2200 Celiac IgA andIgG Kits | Predicate:INOVA QUANTA Liteh-tTG IgA and IgGKits | Predicate:INOVA QUANTA LiteGliadin IgA II and IgGII Kits | |||||
| Antibody | murine monoclonal anti-human IgA or IgG | or IgA HRP-conjugated antibodysolution | or IgA HRP-conjugated antibodysolution | |||||
| Substrate | None | Tetramethylbenzidine(TMB) Chromogen | Tetramethylbenzidine(TMB) Chromogen | |||||
| Signal Detection | Fluorescence | Color, read at 450nm | Color, read at 450nm | |||||
| Solid Phase | Antigen-coated paramagneticmicrobead reagent | Antigen-coated solidphase microtiterwells | Antigen-coated solidphase microtiter wells | |||||
| Calibrator(s) | 5 levels of Calibrator for bothIgA and IgG | 1 level of LowPositive control | l level of LowPositive control | |||||
| Control | One Negative and onePositive Control | One Negative, onelow Positive, and onehigh PositiveControls | One Negative, onelow Positive, and onehigh PositiveControls | |||||
| Assay range | Anti-tTG:IgA: 0.5 to 250 U/mLlgG: 0.8 to 250 U/mLAnti-DGP:IgA: 0.2 to 250 U/mLIgG: 0.4 to 250 U/mL | Not Applicable | Not Applicable | |||||
| Calibrators andControls | Sold Separately | Kit components | Kit components | |||||
| Quantitation | Results are determined froma standard calibration curveutilizing a 4-PL (4-ParameterLogistic) curve fitting. | One point calculationfrom the OD of thelow positive control. | One point calculationfrom the OD of thelow positive control. | |||||
| Instrumentation | Bio-Rad BioPlex 2200system | 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
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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):
-
- 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 | MeanU/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 | MeanU/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 | MeanU/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 | BetweenInstrument/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 | BetweenInstrument/Operator | Between Lot | Total | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample ID | MeanU/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 | BetweenInstrument/Operator | Between Lot | Total | ||||||||
| Sample ID | MeanU/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
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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 CeliacAssay | 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 CeliacAssay | 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 ReactiveDisease 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
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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 withIgA 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'sDisease | 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 -
- Clinical cut-off:
See Assay Cutoff
- Clinical cut-off:
-
- 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.
- Expected values/Reference range:
| 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 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
{22}------------------------------------------------
Page 2-Dr. Juang Wang
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 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 Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 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 Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm.
Sincerely yours,
Maria M. Chan -S
Maria M. Chan, Ph.D. Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
{23}------------------------------------------------
Indications for Use
510(k) Number (if known) K130053
Device Name BioPlex® 2200 Celiac IgA Kit
Indications for Use (Describe)
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 Transglutaminasc (1TG) in human serum. In conjunction with clinical findings and other diagnostic tests, the test system is used as an aid in the disease (gluten-sensitive enteropathy).
The BioPlex 2200 Celiac IgA kit is intended for use with the Bio-Rad BioPlex 2200 System.
Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 807 Subpart C)
PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.
FOR FDA USE ONLY
Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)
Maria M.
1995
FORM FDA 3881 (6/13)
Form Approved: OMB No. 0910-0120 Expiration Date: December 31, 2013 See PRA Statement on last page.
{24}------------------------------------------------
Indications for Use
510(k) Number (if known) K130053
Device Name
BioPlex® 2200 Celiac IgA Control Set
Indications for Use (Describe)
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 the BioPlex 2200 Celiac IgA Reagent Pack in the clinical laboratory. The performance of the BioPlex 2200 Celiac IgA Control Set has not been established with any other anti-tissue Transglutaminase (ITG) and anti-deamidated gliadin peptide IgA assays.
Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)
[] Over-The-Counter Use (21 CFR 807 Subpart C)
PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.
FOR FDA USE ONLY
Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)
Maria M. Chan -S
FDA
FORM FDA 3881 (6/13)
PSC Publishing Services (301) 443-6740 EP
Form Approved: OMB No. 0910-0120 Expiration Date: December 31, 2013 See PRA Statement on last page.
{25}------------------------------------------------
Indications for Use
510(k) Number (if known) K130053
Device Name
BioPlex® 2200 Celiac IgA Calibrator Set
Indications for Use (Describe)
The BioPlex 2200 Celiac IgA Calibrator Set is intended for the BioPlex 2200 Celiac IgA Reagent Pack.
Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)
_ Over-The-Counter Use (21 CFR 807 Subpart C)
PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.
FOR FDA USE ONLY
Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)
Maria M. Chan -
EXA
FORM FDA 3881 (6/13)
Form Approved: OMB No. 0910-0120 Expiration Date: December 31, 2013 See PRA Statement on last page.
PSC Publishing Services (3D) 443-6740 EF
{26}------------------------------------------------
Indications for Use
510(k) Number (if known) K130053
Device Name
BioPlex® 2200 Celiac IgG Control Set
Indications for Use (Describe)
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 the BioPlex 2200 Celiac IgG Reagent Pack in the clinical laboratory. The performance of the BioPlex 2200 Celiac IgG Control Set has not been established with any other anti-tissue Transglutaminase (ITG) and anti-deamidated gliadin peptide IgG assays.
Type of Use (Select one or both, as applicable) .
| X Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 807 Subpart C)
PLEASE DO NOT WRITE BELOW THIS LINE – CONTINUE ON A SEPARATE PAGE IF NEEDED.
FOR FDA USE ONLY
Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)
Maria M. Chan -
FDA
FORM FDA 3881 (6/13)
Form Approved: OMB No. 0910-0120 Expiration Date: December 31, 2013 See PRA Statement on last page.
{27}------------------------------------------------
Indications for Use
510(k) Number (if known) K130053
Device Name
BioPlex® 2200 Celiac IgG Calibrator Set
Indications for Use (Describe)
The BioPlex 2200 Celiac IgG Calibrator Set is intended for the BioPlex 2200 Celiac IgG Reagent Pack.
Type of Use (Select one or both, as applicable)
|x| Prescription Use (Part 21 CFR 801 Subpart D)
[ ] Over-The-Counter Use (21 CFR 807 Subpart C)
PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.
FOR FDA USE ONLY
Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)
Maria M. Chan
FDA
FORM FDA 3881 (6/13)
Form Approved: OMB No. 0910-0120 Expiration Date: December 31, 2013 See PRA Statement on last page.
{28}------------------------------------------------
Indications for Use
510(k) Number (if known) K130053
Device Name BioPlex® 2200 Celiac IgG Kit
Indications for Use (Describe)
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 (1TG) in human serum. In conjunction with clinical findings and other diagnostic tests, the test system is used as an aid in the disease (gluter-sensitive enteropathy).
The BioPlex 2200 Celiac IgG kit is intended for use with the Bio-Rad BioPlex 2200 System.
Type of Use (Select one or both, as applicable)
| > Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 807 Subpart C)
PLEASE DO NOT WRITE BELOW THIS LINE -- CONTINUE ON A SEPARATE PAGE IF NEEDED.
FOR FDA USE ONLY
Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)
Maria M. Char
FDA
FORM FDA 3881 (6/13)
PSC Publishing Services (3(1) 443-6740
§ 866.5660 Multiple autoantibodies immunological test system.
(a)
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).(b)
Classification. Class II (performance standards).