QUANTA PLEX CELIAC IGA PROFILE

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY A. 510(k) Number: k063818 B. Purpose for Submission: New Device C. Measurand: IgA Anti-human tissue transglutaminase (htTG) antibody IgA Anti-deaminated gliadin peptide (DGP) antibody D. Type of Test: Semi-quantitative ELISA E. Applicant: INOVA Diagnostics, Inc. F. Proprietary and Established Names: QUANTA Plex™ Celiac IgA Profile G. Regulatory Information: 1. Regulation section: 21 § CFR 866.5660 Multiple autoantibodies immunological test system 21 § CFR 866.5750 Radioallergosorbent (RAST) immunological test system 2. Classification: II 3. Product code: MVM, Autoantibodies, Endomysial (Tissue Transglutaminase) MST, Antibodies, Gliadin Panel: Immunology 82 H. Intended Use: 1. Intended use(s): The QUANTA Plex™ Celiac IgA Profile is a fluorescent immunoassay for the semi-quantitative detection of IgA anti-human tissue transglutaminase (htTG) and anti-deaminated gliadin peptide (DGP) antibodies, and the detection of an insufficient amount of human serum IgA. The presence of these antibodies in conjunction with other laboratory and clinical findings is an aid in the diagnosis of the gluten sensitive enteropathy celiac disease. Insufficient IgA indicates that there is not enough IgA to allow detection of IgA anti-htTG or anti-DGP. 2. Indication(s) for use: Same as intended use. 3. Special conditions for use statement(s): For prescription use only. 4. Special instrument requirements: Luminex™ laser flow analyzer (Luminex 100 and 200) Luminex™ Integrated System (IS) software program I. Device Description: {1} Each device contains the following: polystyrene microwell plate, 12 (1x8) microwell strips with holder, each microwell contains 4 different colored beadsets and each beadset is coated either purified htTG, DGP, anti-IgA, or IgA; positive and negative controls, Celiac calibrators (human serum IgA antibodies to htTG and DGP antigens); HRP sample diluent; fluorescent-labeled IgA conjugate (goat anti-human IgA alpha chain specific) and conjugate diluent. # J. Substantial Equivalence Information: 1. Predicate device name(s): QUANTA LiteTM htTG IgA QUANTA LiteTM Gliadin IgA II 2. Predicate 510(k) number(s): k011566 (IgA) k052143 (IgA) 3. Comparison with predicate: | Similarities | | | | | --- | --- | --- | --- | | Item | New Device | Predicate Device | | | | QUANTA PlexTM Celiac IgA Profile | QUANTA LiteTM htTG IgA | QUANTA LiteTM Gliadin IgA II | | Antigen | Recombinant htTG and purified synthetic deaminated gliadin peptide | Recombinant htTG | Purified synthetic deaminated gliadin peptide | | Measurement | Semi-quantitative | Same | Same | | Sample | Serum | Same | Same | | Positive and Negative Control | Pre-diluted human serum. Ready to use. | Same | Same | | Sample volume required | 5 μL | Same | Same | | Diluent | HRP sample diluent | Same | Same | | Differences | | | | | --- | --- | --- | --- | | Item | Device | Predicate | | | | QUANTA PlexTM Celiac IgA Profile | QUANTA LiteTM htTG IgA | QUANTA LiteTM Gliadin IgA II | | Intended use | For the semi-quantitative detection of IgA antibodies to recombinant human htTG and synthetic DGP in human serum | For the semi-quantitative detection of IgA antibodies to recombinant human htTG in human serum | for the semi-quantitative detection of IgA antibodies to synthetic DGP in human serum | | Indications for Use | Aid in the diagnosis of both IgA | Aid in the diagnosis of celiac | Aid in the diagnosis of celiac | {2} | Differences | | | | | --- | --- | --- | --- | | Item | Device | Predicate | | | | sufficient and IgA deficient celiac disease | disease and dermatitis herpetiformis | disease | | Technology | Flow cytometer based | ELISA | ELISA | | Assay Format | Multiplexed | Individual analytes | Individual analytes | | Assay Platform | 96 microtiter wells with four differently color-coded sets of antigen coated microspheres | 96 well microtiter plates coated with specific antigen | 96 well microtiter plates coated with specific antigen | | Conjugate | Fluorescent Goat anti-human IgA (alpha chain specific) | Horseradish Peroxidase, Goat anti-human IgA | Horseradish Peroxidase, Goat anti-human IgA | | Assay washing step | None | Two steps | Two steps | | Reading | Luminometer | Spectrophotometer | Spectrophotometer | | Detection Method | Fluorescence | Colorimetric | Colorimetric | | Cut-off | 20.0 LU | 20.0 units | 20.0 units | | Dynamic range | 20 to >20,000 FU | 200 to 300 | 200 to 300 | K. Standard/Guidance Document Referenced (if applicable): None referenced. L. Test Principle: Recombinant htTG and synthetic DGP, anti-IgA and IgA are coupled to different fluorescently 'colored' beadsets. These beadsets are mixed together and put into wells of a microwell plate under conditions that will preserve the antigens in their reactive state. Pre-diluted controls, a serum free control and diluted patient sera are added to separate microwells. If specific antibodies are present, they will bind to the antigen specific beads and free IgA will bind to the anti-IgA beads. Then an anti-human IgA fluorescent conjugated is added to each microwell. A second incubation allows the anti-human IgA fluorescent conjugate to bind to any patient antibodies that are bound to the antigen coated beads or the anti-IgA bead, and the IgA bead. The samples are then analyzed by the Luminex™ flow analyzer. The flow analyzer can distinguish each beadset based on the fluorescent spectrum as well as measure the fluorescent intensity of the conjugate on each bead. The fluorescent intensity on the bead is proportional to the amount of bound conjugate, which in turn is proportional to the amount of patient antibodies bound to the antigen coated beads or IgA bound to the anti-IgA bead. Each antibody can be semi-quantitated by comparing the fluorescent intensity of the patient sample with that of the corresponding Calibrator. Comparing the fluorescent intensities of the anti-IgA bead and the IgA bead can {3} identify the selective IgA-deficient serum as described in the Calculation of Results. These two control beads can also be used to ensure that false negative results due to operator error such as no patient sample added to the well are detected. ## M. Performance Characteristics (if/when applicable): ### 1. Analytical performance: #### a. Precision/Reproducibility: The intra-assay precision was determined by testing eighteen serum samples ten times on a single assay for two different lot numbers of microsphere beads. The inter-assay precision was determined by testing twenty serum samples six times for six days. Results are summarized below. | Intra-assay | | | | | | --- | --- | --- | --- | --- | | | Anti-htTG IgA | Anti-DGP IgA | Anti-IgA | IgA | | Mean LU | 3.0 – 332.0 | 3.0 – 69.0 | 4.7 – 20.6 | 9.5 – 27.9 | | SD | 0.8 – 13.8 | 1.2 – 7.0 | 0.2 – 1.4 | 0.3 – 1.2 | | CV% | 2.2 – 9.3 | 3.2 – 12.0 | 2.1 – 7.4 | 1.1 – 5.1 | | Inter-assay | | | | | | Mean LU | 2.0 – 484.0 | 3.0 – 99.0 | 0.9 – 22.1 | 9.9 – 29.1 | | SD | 0.4 – 17.2 | 0.8 – 10.2 | 0.2 – 1.3 | 0.2 – 2.5 | | CV% | 0.5 – 11.2 | 2.2 – 17.4 | 1.0 – 21.7 | 1.3 – 9.8 | #### b. Linearity/assay reportable range: Not applicable. #### c. Traceability, Stability, Expected values (controls, calibrators, or methods): There are no reference standards for htTG and DGP. The positive and negative controls are prepared in-house and arbitrary units are assigned during the development process. Stability: The expiration date claims are one year for the QUANTA Plex™ Celiac IgA Profile kit and three months for the reconstituted IgA conjugate. #### d. Detection limit: The detection limit was determined by testing serially diluted low positive patient serum. Values did not decrease below 4 LU on further dilutions. The anti-htTG or anti-DGP antibodies were detectable at 4 LU. #### e. Analytical specificity: Interference by endogenous substances: A total of six serum samples were tested. One sample was used for each of the following interferents: 1000 mg/dL hemoglobin, 29.7 mg/dL bilirubin, 369 mg/dL cholesterol, 1016 mg/dL triglycerides and 5 µg/mL IgA on equal volumes of positive anti-htTG and anti-DGP. No or negligible interference was observed. The package insert states that grossly hemolyzed, lipemic, microbially contaminated, heat-treated samples or specimens containing visible particulate should not be used in this assay. Crossreactivity with other autoantibodies: The QUANTA Plex™ Celiac IgA {4} Profile was tested with 63 sera consisting of 10 RF and/or anti-CCP, 11 ANA IFA positive and 42 infectious disease samples positive for HCV, HSV, CMV, Toxoplasmosis, rubella or parvovirus. All samples were negative with the QUANTA Plex™ Celiac IgA Profile. f. Assay cut-off: The cut-off value of 20 LU for the assay was established from 278 asymptomatic blood donors. Age and gender were available for 38 samples and unavailable for the remaining 240 samples. The assay specificity was 99.6% (277/278). The cut-off value of arbitrary LU was chosen for the continuity of INOVA products. The value of the fluorescence that was assigned 20 LU was based on a non-parametric statistical analysis of the data. 2. Comparison studies: a. Method comparison with predicate: Testing was performed on 954 samples (278 asymptomatic blood donors, 29 Celiac Disease (CD) IgA sufficient patients, and 647 samples with other disease states or conditions. The 647 samples includes: 53 GI and liver diseases; 63 infectious and rheumatic diseases; 10 autoimmune thyroid diseases; 4 SLE; 22 RA; 18 1st degree CD relative; 39 defined IgA samples; 44 CD [32 GFD, 7 suspected and 5 selected IgA deficient]; and 394 samples with no CD clinical symptoms but may or may not have antibodies to CD. The comparative study on anti-htTG IgA had a Positive Percent Agreement of 89.2% (174/195); Negative Percent Agreement 99.7% (757/759) and Overall Agreement 97.6% (931/954) (refer to table below). | Anti-htTG IgA | QUANTA Lite™ htTG IgA (Elisa) | | | | | --- | --- | --- | --- | --- | | | | Positive | Negative | Total | | QUANTA Plex™ Celiac IgA Profile (Luminex) | Positive | 174 | 2 | 176 | | | Negative | 21 | 757 | 778 | | | Total | 195 | 759 | 954 | The comparative study on anti-DGP IgA had a Positive Percent Agreement of 81.4% (136/167); Negative Percent Agreement of 99.5% (783/787) and Overall Agreement of 96.3% (919/954) (refer to table below). | Anti-DGP IgA | QUANTA Lite™ Gliadin IgA II (Elisa) | | | | | --- | --- | --- | --- | --- | | | | Positive | Negative | Total | | QUANTA Plex™ Celiac IgA Profile (Luminex) | Positive | 136 | 4 | 140 | | | Negative | 31 | 783 | 814 | | | Total | 167 | 787 | 954 | Quanta Plex Celiac IgA on Luminex 200 and IS.v.2.3 software For this study, 96 samples were tested and results were compared to the Luminex 100. Results were as follows: {5} 6 | Anti-htTG IgA | Luminex 100 | | | | | --- | --- | --- | --- | --- | | | | positive | negative | Total | | Luminex 200 | positive | 40 | 0 | 40 | | | negative | 0 | 56 | 56 | | | Total | 40 | 56 | 96 | Positive percent agreement 100% (40/40) Negative percent agreement 100% (56/56) Overall percent Agreement 100% (96/96) | Anti-DGP IgA | Luminex 100 | | | | | --- | --- | --- | --- | --- | | | | positive | negative | Total | | Luminex 200 | positive | 23 | 0 | 23 | | | negative | 1 | 72 | 73 | | | Total | 24 | 72 | 96 | Positive percent agreement 96% (23/24) Negative percent agreement 100% (72/72) Overall percent Agreement 99% (95/96) b. Matrix comparison: Both assays use serum as the matrix. 3. Clinical studies: a. Clinical Sensitivity and Specificity: The study consisted of the following: 29 samples from active CD IgA sufficient patients and 494 samples not diagnosed with CD. The 494 samples include the following: 278 asymptomatic blood donors; 53 GI/liver disease; 42 infectious diseases (positive for HCV, HSV, CMV, Toxoplasmosis, rubella, or parvovirus); 11 ANA IFA Positive; 10 RF and/or anti-CCP Positive; 93 autoimmune diseases (10 thyroid disease, 4 SLE, 22 RA, 18 1st degree CD relative, 39 defined IgA sufficient CD patients); and 7 suspected CD. The clinical study on anti-htTG IgA were as follows: clinical sensitivity was 93.1% (27/29); clinical specificity was 97.8% (483/494) and overall agreement was 97.5% (510/523) (refer to table below). | Anti-htTG IgA | Diagnosis | | | | | --- | --- | --- | --- | --- | | | | Positive (CD with sufficient IgA and not on GFD) | Negative (Not diagnosed with CD) | Total | | QUANTA Plex™ Celiac IgA Profile (Luminex) | Positive | 27 | 11** | 38 | | | Negative | 2* | 483 | 485 | | | Total | 29 | 494 | 523 | * Both samples were Elisa anti-htTG IgA negative. ** Ten of these samples were Elisa htTG IgA positive {6} The clinical study on anti-DGP IgA were as follows: clinical sensitivity was 65.5% (18/29); clinical specificity was 97.6% (482/494) and overall agreement was 95.6% (500/523) (refer to table below). | Anti-DGP IgA | Diagnosis | | | | | --- | --- | --- | --- | --- | | | | Positive | Negative | Total | | QUANTA Plex™ Celiac IgA Profile (Luminex) | Positive | 18 | 12** | 31 | | | Negative | 11* | 482 | 492 | | | Total | 29 | 494 | 523 | *Ten of these samples were Elisa anti-DGP IgA negative. **Ten of these samples were Elisa anti-DGP IgA positive. b. Other clinical supportive data (when a .is not applicable): Not applicable. 4. Clinical cut-off: Same as assay cut-off. 5. Expected values/Reference range: Expected values in the normal population should be negative. N. Proposed Labeling: The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10. O. Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision.