(90 days)
The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is an in vitro diagnostic test for the detection and genotyping of the *2 and *3 CYP4502C9 genetic variants and the VKORC1 3673 (-1639) intronic variant in genomic deoxyribonucleic acid (DNA) obtained from EDTA-anticoagulated whole blood samples. The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is a qualitative assay for use in clinical laboratories upon prescription by the attending physician.
The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is indicated for use to identify individuals at risk for sensitivity to warfarin.
The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is an in vitro diagnostic device which utilizes proprietary film-based microarray technology combined with process automation, reagent management, and software technology for the detection and genotyping of the 2C92, 2C93, and VKORC1 3673 (-1639) mutations from EDTA-anticoagulated whole blood samples.
The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is comprised of the BioFilmChip™ Microarray, the Intellipac Reagent Module and the PCR Amplification Mix. The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin should be run using the AutoGenomics INFINITI Analyzer.
The BioFilmChip Microarray consists of a polyester film coated with proprietary multi-layer components designed for DNA analysis. The layers have been designed to provide a versatile surface to enhance test performance. There can be up to 240 spots per microarray with each spot representing a different allele. The microarrays are designed to be assay specific.
The Intellipac Reagent Module contains up to eight reservoirs that house the test reagents and has an integrated memory chip. Information on the reagent such as lot number, expiration date and volume usage, are archived in the memory.
The PCR Amplification Mix consists of the reagents needed for the PCR amplification step of the assay.
The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is based on the following processes:
(a) DNA extraction
(b) PCR amplification of purified DNA from human genomic DNA
(c) Labeling of the amplified product (allele specific primer extension)
(d) Hybridization of the labeled amplified product to a microarray by signature Tag/Capture probe hybridization under isothermal conditions.
(e) Scanning of the microarray
(f) Signal detection and analysis [determination of the 2C92, 2C93 and VKORC1 3673 (-1639) genotypes]
Steps (c) through (f) are automated by the INFINITI Analyzer.
The INFINITI Analyzer automates the 2C9 and VKORC1 assays and integrates all the discrete processes of sample (PCR amplicon) handling, reagent management, hybridization, detection, and results analysis. The assays are processed automatically and read by the built-in confocal microscope. Results are analyzed and presented as genotype calls.
Here's an analysis of the acceptance criteria and study details for the INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin, based on the provided text:
Acceptance Criteria and Device Performance
The document describes the performance characteristics without explicitly stating pre-defined "acceptance criteria" as pass/fail thresholds. Instead, it presents the "reported device performance" and implies that these results demonstrate the device's suitability. For the purpose of this analysis, I will synthesize the reported performance values as the de facto "acceptance criteria" that the device met.
1. Table of Acceptance Criteria and Reported Device Performance
| Performance Metric | Acceptance Criteria (Implied) | Reported Device Performance |
|---|---|---|
| Analytical Specificity | PCR primer, ASP primer, and capture probe specificity confirmed. | PCR primer specificity by amplicon size & sequencing. ASP primer specificity by correct calls with known genomic samples. Capture probe specificity by hybridization of correct oligo to known spot. |
| Limit of Detection (Analytical Sensitivity) | Minimum DNA concentration for correct calls. Recommended DNA concentration for reliable results. | Minimum DNA concentration for correct calls: 1 ng DNA. Recommended DNA concentration: 25 ng/µl (50 ng per test). Correct calls at 10 ng DNA. |
| Agreement with Bi-directional Sequencing (1st run) | High percentage agreement for *2, *3, and VKORC1 variants. | 2C9*2: 98.0% agreement (95% CI Lower Limit: 95.09%) |
| 2C9*3: 97.3% agreement (95% CI Lower Limit: 94.09%) | ||
| VKORC1 3673 (-1639): 98.0% agreement (95% CI Lower Limit: 95.09%) | ||
| Overall assay agreement. | Total for Assay: 97.8% agreement (95% CI Lower Limit: 96.86%) | |
| Agreement with Bi-directional Sequencing (Final Result - after re-run of no calls) | High percentage agreement, minimizing no calls and incorrect calls. | Total for Assay: 99.3% correct call rate (149/150 samples) for Table 2b (by Sample Type) |
| Assay Inter-Laboratory Reproducibility (Genotype Call Rate) | High percentage of correct genotype calls across sites, within each genotype, and overall. | 2C9*2 (Total): 99.07% (98.06% 95% CI Lower Limit) |
| 2C9*3 (Total): 97.22% (90.47% 95% CI Lower Limit) | ||
| VKORC1 3673 (-1639) GA (Total): 99.44% (98.08% 95% CI Lower Limit) | ||
| VKORC1 3673 (-1639) AA (Total): 97.22% (92.73% 95% CI Lower Limit) | ||
| VKORC1 3673 (-1639) GG (Total): 100.0% (98.08% 95% CI Lower Limit) | ||
| Total for Assay (Reproducibility): 98.99% correct calls (98.42% 95% CI Lower Limit) after 1st run; 99.96% after repeat. | ||
| Drug Interference | No interference from common interfering substances. | No interference from bilirubin (8mg/dl), cholesterol (70mg/dl), and heparin (133v/dl). |
| Sample Carry-Over | No detection of sample carry-over. | 100% correct genotype calls when positive sample followed by another positive or "No Template Control". |
| Assay Interference | No interference between this assay and other assays on the same instrument. | No interference when run simultaneously with INFINITI Assay for Factor II & Factor V. |
| Reagent Stability | Demonstrated shelf-life at specified storage conditions. | BioFilmChip Microarray: 12 months at RT (15-30°C). Intellipac Reagent: 12 months Refrigerated (2-8°C). Amplification Mix: 12 months Frozen (-10°C). |
2. Sample Sizes Used for the Test Set and Data Provenance
Sample Size for Test Set:
- Agreement with Bi-directional Sequencing:
- 150 patient samples tested in the initial comparison (first run data for 2C9*2, 2C9*3, VKORC1).
- Table 2b "by Sample Type" also shows 150 samples, implying these are the same samples analyzed by genotype.
- Inter-Laboratory Reproducibility:
- 7 genomic DNA samples and 5 whole blood samples (total 12 unique samples).
- Each unique sample was run in duplicate per day/operator for six days at each of 3 sites.
- Total tests: 12 samples * 2 replicates/day * 6 days * 3 sites = 432 tests per site, for a total of 1296 tests across all sites for the "first time run" and "final result".
Data Provenance:
- Agreement with Bi-directional Sequencing:
- Origin: Not explicitly stated, but "patient samples" were used. Given the FDA submission context, it's highly likely these were de-identified samples from the United States.
- Nature: The description "Each site tested its own patient samples" and that they were "from patients using or have used warfarin" suggests these were retrospective samples, collected from a patient population relevant to the intended use.
- Inter-Laboratory Reproducibility:
- Origin: Not explicitly stated, but likely from the United States given the submission.
- Nature: Controlled study using "seven genomic DNA samples and five whole blood samples." These would be prospective in the sense that they were specifically prepared and distributed for this reproducibility study, though the original source of the genetic material might have been retrospective.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications
The document does not mention the use of experts to establish ground truth.
For the Agreement with Bi-directional Sequencing study, the ground truth was established by:
- Bi-directional DNA sequencing. This is a recognized laboratory method for determining genetic sequences and is considered the gold standard for many genetic variants. Thus, the ground truth was established by laboratory method, not expert consensus.
- The document implies that the sequencing results were accepted as the definitive truth without the need for expert adjudication or review.
For the Inter-Laboratory Reproducibility study, the ground truth for the 7 genomic DNA samples and 5 whole blood samples was their known genotypes, which would have been previously determined, presumably also by a method like bi-directional sequencing.
4. Adjudication Method for the Test Set
The document does not describe an adjudication method involving multiple human reviewers for the test set.
- In the "Agreement with Bi-directional Sequencing" study, "bi-directional sequencing" served as the comparator/ground truth. The device results were directly compared to these sequencing results.
- For the inter-laboratory reproducibility study, device results from different sites and operators were compared to the known genotype of the samples.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
No MRMC comparative effectiveness study was done. This study assesses the improvement of human readers with AI assistance versus without AI assistance. The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is an in vitro diagnostic device for genotype detection, not an imaging or diagnostic support tool that assists human readers. Its output is a genotype call, not an interpretation that radiologists or other human experts would then refine or improve upon.
6. Standalone Performance Study (Algorithm Only)
Yes, a standalone performance study was done. The entire submission details the performance of the "INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin" as a standalone device. The device automates key steps and provides genotype calls:
- "Steps (c) through (f) are automated by the INFINITI Analyzer."
- "The INFINITI Analyzer automates the 2C9 and VKORC1 assays and integrates all the discrete processes of sample (PCR amplicon) handling, reagent management, hybridization, detection, and results analysis. The assays are processed automatically and read by the built-in confocal microscope. Results are analyzed and presented as genotype calls."
- The performance metrics (agreement with sequencing, reproducibility, LOD, specificity, etc.) are all measures of the algorithm's direct output.
There is no human-in-the-loop component described for its routine operation or performance evaluation.
7. Type of Ground Truth Used
The primary ground truth used for the performance studies was established laboratory method results, specifically bi-directional DNA sequencing. This method is considered highly accurate for determining DNA sequences and genetic variants.
8. Sample Size for the Training Set
The document does not specify a sample size for a training set. This is typical for in vitro diagnostic devices based on established molecular biology principles (PCR, hybridization, genotyping microarrays) where the design is more mechanistic and less dependent on machine learning models requiring extensive "training data" in the conventional sense. The "development" of the assay mentioned for analytical specificity would involve testing various known samples, but these are generally considered part of the assay development and validation rather than a distinct "training set" for an algorithm in the AI context.
9. How the Ground Truth for the Training Set Was Established
Since no explicit training set is detailed, the method for establishing its ground truth is also not described. If one were to consider the samples used during "assay development" as a form of training/optimization, their ground truth would also have been established by known genomic samples (e.g., cell lines with characterized genotypes, or samples sequenced by a gold-standard method).
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K073014
510(K) SUMMARY
。
| Submitted By | AutoGenomics, Inc.2251 Rutherford RoadCarlsbad, CA 92008, USATelephone: (760) 804-7378FAX: (760) 804-7382 | JAN 23 2008 | |
|---|---|---|---|
| Contact: Evelyn LopezVice President, Regulatory Affairs | |||
| Date Prepared: January 11, 2008 | |||
| Device Name | Trade or Proprietary Name: INFINITI 2C9 & VKORC1 Multiplex Assay for WarfarinCommon or Usual Name: 2C9 and VKORC1 Drug Metabolizing Enzyme Genotyping System | ||
| Regulations andProduct Codes | Regulations: 21CFR§862.3360 Drug Metabolizing Enzyme Genotyping Systems21CFR§864.7750 Prothrombin Time Test21CFR§862.2570 Instrumentation for Clinical Multiplex Test SystemsProduct Codes: ODW Cytochrome P450 2C9 (CYP450 2C9) Drug Metabolizing Enzyme Genotyping SystemODV Vitamin K epoxide reductase complex subunit 1 (VKORC1) Genotyping SystemNSU Instrumentation for Clinical Multiplex Test Systems | ||
| Predicate Device | (a) AmpliChip CYP450 Test for CYP2C19(b) INFINITI System Assay for Factor II & Factor V | ||
| Device Description | The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is an in vitro diagnostic device which utilizes proprietary film-based microarray technology combined with process automation, reagent management, and software technology for the detection and genotyping of the 2C92, 2C93, and VKORC1 3673 (-1639) mutations from EDTA-anticoagulated whole blood samples.The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is comprised of the BioFilmChip™ Microarray, the Intellipac Reagent Module and the PCR Amplification Mix. The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin should be run using the AutoGenomics INFINITI Analyzer.The BioFilmChip Microarray consists of a polyester film coated with proprietary multi-layer components designed for DNA analysis. The layers have been designed to provide a versatile surface to enhance test performance. There can be up to 240 spots per microarray with each spot representing a different allele. The microarrays are designed to be assay specific.The Intellipac Reagent Module contains up to eight reservoirs that house the test reagents and has an integrated memory chip. Information on the reagent such as lot number, expiration date and volume usage, are archived in the memory. |
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| The PCR Amplification Mix consists of the reagents needed for the PCR amplificationstep of the assay. | |
|---|---|
| The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is based on the followingprocesses:(a) DNA extraction(b) PCR amplification of purified DNA from human genomic DNA(c) Labeling of the amplified product (allele specific primer extension)(d) Hybridization of the labeled amplified product to a microarray by signatureTag/Capture probe hybridization under isothermal conditions.(e) Scanning of the microarray(f) Signal detection and analysis [determination of the 2C92, 2C93 and VKORC13673 (-1639) genotypes]Steps (c) through (f) are automated by the INFINITI Analyzer. | |
| The INFINITI Analyzer automates the 2C9 and VKORC1 assays and integrates all thediscrete processes of sample (PCR amplicon) handling, reagent management,hybridization, detection, and results analysis. The assays are processed automatically andread by the built-in confocal microscope. Results are analyzed and presented as genotypecalls. | |
| Intended Use | The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is an in vitro diagnostictest for the detection and genotyping of the *2 and *3 CYP4502C9 genetic variants andthe VKORC1 3673 (-1639) intronic variant in genomic deoxyribonucleic acid (DNA)obtained from EDTA-anticoagulated whole blood samples. The INFINITI 2C9 &VKORCI Multiplex Assay for Warfarin is a qualitative assay for use in clinicallaboratories upon prescription by the attending physician. |
| Indication for Use | The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is indicated for use toidentify individuals at risk for sensitivity to warfarin. |
| SubstantialEquivalence | Tables 1a and 1b provide a comparison of the technologicalcharacteristics of the INFINTI 2C9 & VKORC1 Multiplex Assay for Warfarin to those ofthe predicate devices• AmpliChip CYP450 Test for CYP2C19 (K043576) |
:
- . . INFINITI System Assay for Factor II & Factor V (K060564)
Table 1a
| Characteristics | PredicateAmpliChip CYP450 Test for CYP2C19 | Subject DeviceINFINITI 2C9 & VKORC1Multiplex Assay for Warfarin |
|---|---|---|
| Similarities | ||
| DNA sequence | Detects specific DNA sequences through recognition of DNA targets | Same |
| Technology | Microarray-based genotyping test for simultaneous detection(multiplex system) of DNA sequences | Same |
| Specimen Type | Purified DNA from human blood samples | Same |
| Reaction Conditions | Utilizes thermal cycling Utilizes target DNA amplification | Same |
| Assay Results | Assay signal results are interpreted by a software program Assay results are provided as genotype calls reported to the end user in a report format | Same |
| Differences | ||
| Gene | CYP2D6 and CYP2C19 | CYP2C9 and VKORC1 |
| Microarray substrate | Reactions occur on a single glass slide | Reactions occur on a single biofilmmicroarray chip |
| Number of alleles | 2 | 3 |
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Table 1b
| Predicate | Subject Device | |
|---|---|---|
| Characteristics | INFINITI System Assay forFactor II & Factor V | INFINITI 2C9 & VKORC1Multiplex Assay for Warfarin |
| Similarities | ||
| DNA sequence | Detects specific DNA sequences through recognition of DNA targets | Same |
| Technology | Microarray-based genotyping test for simultaneous detection(multiplex system) of DNA sequences | Same |
| Specimen Type | Purified DNA from human blood samples | Same |
| Reaction Conditions | Utilizes thermal cycling Utilizes target DNA amplification | Same |
| Microarray substrate | Reactions occur on a single biofilm microarray chip | Same |
| Assay Results | Assay signal results are interpreted by a software program Assay results are provided as genotype calls reported to the end user in a report format | Same |
| Differences | ||
| Gene | Factor II and Factor V | CYP2C9 and VKORC1 |
| Number of alleles | 2 | 3 |
Performance
The following are performance characteristics of the INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin:
Analytical Specificity
Studies related to specificity were conducted during assay development. PCR primer specificity was determined by amplicon size on a gel and sequencing the amplicon. ASP primer specificity was determined by the correct calls made by the assay using known genomic samples. Capture probe specificity was determined by hybridizing different oligos and demonstrating that correct oligo hybridizes to the known spot.
Limit of Detection (analytical sensitivity)
Serial dilutions (200, 100, 50, 25, 10, 1, 0.1ng DNA) were prepared from a known purified DNA sample. Each serial dilution was assayed three times using the INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin. The study established the minimum DNA concentration for the INFINITI System Assay for 2C9-VKORC1 to be Ing DNA. The recommended DNA concentration for the INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is 25ng/ul. The assay requires 2ul of DNA sample or the equivalent of 50ng per test.
In addition, the same study demonstrated that DNA concentrations of 100ng and 200ng, which were in excess of the recommended concentration (50ng), did not interfere with the INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin. The following table provides a summary of the results of the study.
| Sample | Run 1 | Run 2 | Run 3 | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| ng DNA | Genotype | Genotype Calls | Result | Genotype Calls | Result | Genotype Calls | Result | |||||||
| 2C9 | VKORCI3673 (-1639) | 2C9*2 | 2C9*3 | VKORCI3673 (-1639) | 2C9*2 | 2C9*3 | VKORCI3673 (-1639) | 2C9*2 | 2C9*3 | VKORCI3673 (-1639) | ||||
| 200 | W | H | W | W | H | Pass | W | W | H | Pass | W | W | H | Pass |
| 100 | W | H | W | W | H | Pass | W | W | H | Pass | W | W | H | Pass |
| 50 | W | H | W | W | H | Pass | W | W | H | Pass | W | W | H | Pass |
| 25 | W | H | W | W | H | Pass | W | W | H | Pass | W | W | H | Pass |
| 10 | W | H | W | W | H | Pass | W | W | H | Pass | W | W | H | Pass |
| 1 | W | H | W | W | H | Pass | W | W | H | Pass | W | W | H | Pass |
| 0.1 | W | H | No call | W | H | Fail | W | W | H | Pass | No call | W | H | Fail |
Limit of Detection
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Percent Agreement vs. Bi-directional Sequencing
The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin was compared to bi-directional sequencing as the comparator method. Three sites were used for the comparison studies. Each site tested its own patient samples with the INFINITI 2C9-VKORC1 Multiplex Assay for Warfarin. All samples used in the comparison studies at the three sites were from patients using or have used warfarin. Patient samples were de-identified to protect patient's identity. Each site performed the DNA extraction using a different extraction method. The results of the comparison studies demonstrated
98.0% agreement for 2C92 as compared with bi-directional sequencing on 1st run 97.3% agreement for 2C93 as compared with bi-directional sequencing on 15 run 98.0% agreement for VKORC1 3673 (-1639) as compared with bi-directional sequencing on 1st run
The results of the comparison studies are summarized in Table 2a and Table 2b.
Table 2a Agreement between INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin and Bi-Directional DNA Sequencing
| Genotype" | NumberTested | Replicatesper Sample | Number ofCorrectGenotype Calls" | Number ofIncorrect Calls | No Calls | Agreement | 95% One-SidedConfidenceLowerLimit |
|---|---|---|---|---|---|---|---|
| 2C9*2*1/*2 | 35 | 1 | 34 | 0 | 1 | 97.1% | 88.82% |
| 2C9*2*2/*2 | 2 | 1 | 2 | 0 | 0 | 100.0% | 50%d |
| 2C9*2*1/*1 | 113 | 1 | 111 | 0 | 2 | 98.2% | 94.91% |
| Total for 2C9*2 | 150 | 1 | 147 | 0 | 3 | 98.0% | 95.09% |
| 2C9*3*1/*3 | 19 | 1 | 19 | 0 | 0 | 100.0% | 80.45% |
| 2C9*3*3/*3 | 1 | 1 | 1 | 0 | 0 | 100.0% | 0%d |
| 2C9*3*1/*1 | 130 | 1 | 126 | 1c | 3 | 96.9% | 94.30% |
| Total for 2C9*3 | 150 | 1 | 146 | 1 | 3 | 97.3% | 94.09% |
| VKORCI3673 (-1639)GA | 63 | 1 | 62 | 0 | 1 | 98.4% | 93.74% |
| VKORCI3673(-1639)AA | 27 | 1 | 25 | 0 | 2 | 92.6% | 79.01% |
| VKORCI3673 (-1639)GG | 60 | 1 | 60 | 0 | 0 | 100.0% | 98.33% |
| Total forVKORCI3673 (-1639) | 150 | 1 | 147 | 0 | 3 | 98.0% | 95.09% |
| Total forAssay | 450 | 1 | 440 | 1 | 9 | 97.8% | 96.86% |
a Genotype determined through bi-directional DNA sequencing
b Calls produced on first run
් Initial INFINITI results (*1/1 for 2C92, *1/3 for 2C93 and GG for VKORC1 3673) did not match bi-directional sequence results (*1/1 for 2C92, *1/1 for 2C93 and GG for VKORC1 3673). The same INFINITI results were obtained on repeat run. Reason unknown.
d For sample sizes 1 and 2, and 100% agreement, SE(p2-p1)=0. Pure sample size correction for sample size 2 is 50% and for sample size 1 is 100%, therefore, One-Sided Confidence Lower Limits are 50% (n=2) and 0% (n=1).
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| Sample Description Genotype* | First Time Run | Final Resultd | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| 2C9 | VKORC13673 (-1639) | *2 | *3 | # Samples Tested | Samples with Genotype Calls made by INFINITIb | Samples with Correct Callsc | Samples with No Incorrect Calls | Samples with Genotype Calls made by INFINITIb | Samples with Correct Callsc | Samples with No Incorrect Calls | Correct Call Ratee (%) |
| *1/*1 | ]/] | ]/] | 20 | 18 | 18 | 2 | 20 | 20 | 0 | 90.0 | |
| *1/*1 | *1/*1 | *1/*1 | 34 | 34 | 34 | 0 | 34 | 34 | 0 | 100 | |
| *1/*1 | *1/*1 | *1/*1 | 43 | 43 | 42 | 0 | 43 | 42 | 1 | 97.7 (42/43) | |
| *1/*2 | *1/*1 | *1/*1 | 4 | 4 | 4 | 0 | 4 | 4 | 0 | 100 | |
| *1/*2 | *1/*1 | *1/*1 | 15 | 14 | 14 | 1 | 15 | 15 | 0 | 100 | |
| *1/*2 | *1/*1 | *1/*1 | 12 | 12 | 12 | 0 | 12 | 12 | 0 | 100 | |
| *1/*1 | *1/*3 | *1/*1 | 2 | 2 | 2 | 0 | 2 | 2 | 0 | 100 | |
| *1/*1 | *1/*3 | *1/*1 | 10 | 10 | 10 | 0 | 10 | 10 | 0 | 100 | |
| *1/*1 | *1/*3 | *1/*1 | 3 | 3 | 3 | 0 | 3 | 3 | 0 | 100 | |
| *2/*2 | *1/*1 | *1/*1 | 1 | 1 | 1 | 0 | 1 | 1 | 0 | 100 | |
| *2/*2 | *1/*1 | *1/*1 | 1 | 1 | 1 | 0 | 1 | 1 | 0 | 100 | |
| *1/*2 | *1/*3 | *1/*1 | 1 | 1 | 1 | 0 | 1 | 1 | 0 | 100 | |
| *1/*2 | *1/*3 | *1/*1 | 2 | 2 | 2 | 0 | 2 | 2 | 0 | 100 | |
| *1/*2 | *1/*3 | *1/*1 | 1 | 1 | 1 | 0 | 1 | 1 | 0 | 100 | |
| *1/*1 | *3/*3 | *1/*1 | 1 | 1 | 1 | 0 | 1 | 1 | 0 | 100 | |
| Total | 150 | 147 | 146 | 3 | 150 | 149 | 1 | 99.3 (149/150) |
able 2b Agreement between INFINITI 2C9 & VKORCI Multiplex Assay for Warfarin and Bi-Directional DNA Sequencing (by Sample Type
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Assay Inter-Laboratory Reproducibility
A three-site study was conducted to demonstrate the reproducibility of the INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin. The study involved three identical lots of the INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin. The sites ran identical samples comprised of seven genomic DNA samples and five whole blood samples. The sites were blinded to sample identity. Each site used a different DNA extraction method. At each site, each sample was run in duplicate per day/operator for six days. Three operators were required for each site. Results of the inter-laboratory reproducibility study are summarized in Tables 3a and 3b.
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| ble 3a Inter-Laboratory Reproducibility of the INFINITI 2C9 | First Time Run | KORC1 Multiplex Assay for Warfarin by Genotype ca | inal Result | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| enotyp | Gamples Tested | Tests Per Site | Site | Genotype Calls | 96 | |||||||||
| Correc Calls | Incorrect Calls | נקום קנטו | % Correc | 95% One-Side Confidence Lower Limit | Correct Calls | ncorrec . Calls | ות היום | Correc Calls | 5% One-Side Confidence Lower Limit | |||||
| { | 98 | 98 | 100 | 0 | 0 | 100 | ||||||||
| て #/ # これだと | క్ | 98 | ट | 9E | ਡ | ಲ್ಲಿ | 94.4 | 9 2018 | 0 | 0 | 100 | |||
| દ | 9 દ | 9 ද | 100 | 0 | 1 00 | |||||||||
| tota | 08 | 106 | 0 NO | 98.15 | 95.14% | 108 | 0 0 | 0 | 100 | 99.53% | ||||
| ਟ ไ | ਟ । | 100 | 0 | 0 | 1 (AC | |||||||||
| て #/ぴょ 2 #63 | l | 12 | ದ | ਟ ਦੀ ਨਾ | 11 | ျ l | 91.7 | ત્યાર્ટ | 0 | 0 | 100 | |||
| દ | ਨ। | 0 | 1 00 | 0 | 0 | 100 | ||||||||
| tota | 97 | ਟ ਵਿ | [ | 97.22 | 90.47% | 9 દ | 0 | 100 | 98.61% | |||||
| ໄ | 96 | r I | 99.0 | 96 | 00000 | 0 | 100 | |||||||
| 【 #/ # C #637 | 8 | 96 | ਟ | 96 | તેણ | 0 | 1 00 | 96 | 0 | 100 | ||||
| દ | 96 | 96 | 0 | 100 | 96 | 0 | 100 | |||||||
| ota | 288 | 287 | ໄ | 99.65 | 99.13 | 0 | 100 | 09.83 | ||||||
| otal for 2C9 | 432 | 428 | t | 99.07 | 98.06% | 2017 882 | 0 | 0 | 100 | 99.88 | ||||
| 24 | 100 | 0 | 0 | 100 | ||||||||||
| € * / ½ § * 63 | ಒಂದಿಗಳು ಒಂದು ಒಂದು ಮಾರ್ಥಿಗಳು ಒಂದು ಮಾರ್ಥಿಗಳು ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾರ್ಗ್ ಮಾಡಿ ಮಾರ್ಗ್ ಮಾಡಿ ಮಾರ್ಗ್ ಮಾಡಿ ಮಾರ್ಗ್ ಮಾಡಿ ಮಾರ್ಗ್ ಮಾಡಿ ಮಾರ್ಗ್ ಮಾಡಿ ಮಾರ್ಗ್ ಮಾಡಿ ಮಾರ್ಗ್ ಮಾಡಿ ಮಾರ್ | র ব | ਟ | ਦੇ ਇੱਕ | ਟੂਰ ਦੇ ਇੱਕ | ofic | 91.7 | 0 | 0 | I OO | ||||
| ది | 1 00 | 0 | 0 | 100 | ||||||||||
| tola | 70 | 0 N | 97.22 | 92.73% | 0 | 0 | 100 | 99.30% | ||||||
| ໄ | ਹੈ। ਇਹ | ਲ ਕ | 0 | 100 | 0 | 0 | 100 | |||||||
| E #16 # E#637 | ਟ | র ব | ত্র | ﺳﺎﺗﮫ ﮨﮯ | 100 | 지지만 지지자 지원 | 0 | 0 | 100 | |||||
| દુ | র্যা | ದ್ದ ಪ್ರಸಿದ್ದ ಮತ್ತು ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ ಮಾಡಿ | 05.8 | 0 0 | 0 | 100 | ||||||||
| tota | ਟ ( | 98.6 | 95.21% | 0 ﺍﻟﻤ | 100 | 99.30% | ||||||||
| 96 | SS | g I | 99.0 | 96 | 0 | 100 | ||||||||
| T #/ I * E #637 | 8 | 96 | ત્યાળ | ેણ | તેરે રે | վ մ | 99.0 | 96 | 0 0 | 0 0 | 100 | |||
| ഴു | 982 96 | 0 | 100 | 887 96 | 1 00 | |||||||||
| ota | 288 | 99.3 | 98.17% | 0 | 100 | |||||||||
| otal for 2C9 | 432 | 427 | 2000 | 98.84 | 97.72% | 432 | 0 0 | 0 0 | 1 00 | 99.83% 99.88% | ||||
| 60 | 09 | 100 | 09 | 0 | 100 | |||||||||
| רי | 09 | ਟ | 60 | રતે | 98.3 | 09 | 0 | 0 | 100 | |||||
| VKORCI 673 (-1639 GA | ర్థి మూలాలు కార్యాలయం గ్రామం నుండి 10 కి.మీ. దూరంలో ఉన్నాయి. ప్రాథమిక ఆరోగ్యం మూలాలు విద్యా శిశు సంరక్షణ కార్యాలయం విశాఖపట్నం మూలాలు విశాఖపట్నం మూలాలు విశాఖపట్నం క | 60 | 60 | 0 | 1 00 | 09 | 0 | 0 | 100 | |||||
| tota | 186 | 179 | l | 99.44 | 98.08% | 180 | 0 | 0 | 100 | 99 72% | ||||
| すことです。 あなたに | な | വില് വിവിവിച്ചിരിക്കുന്നതിന് എന്നിവരിക്കുന്നതിന്റെ വിവിധ | 100 | 0 | 0 | 100 | ||||||||
| VKORC1 673 (-1639) | ત્ત્વ | な | ন্ত | ਟ ਟ | 0 % 0 | 91.7 | 22222 | 0 0 | 00 | 100 | ||||
| ਦੀ ਇੱ | । ਡੇ ਟਿ | 1 OC | 100 | |||||||||||
| tota | ไฟ ไม | 97.22 | 92.73 | 0 | 00 | 100 | 99.30% | |||||||
| 60 | 65 | 98.3 | 09 | 0)0 | 100 | |||||||||
| VKORCI 673 (-1639) GG | S | 09 | പ | 09 | 60 | 0000000 | 0 | 100 | 60 | 0 | 100 | |||
| డు | 60 | 60 | ol- | 100 | 09 | 0 0 | 1 00 | |||||||
| tota | 08 I | 179 | 10 66 | 98.08% | 180 | 000 | 100 | 99.72% | ||||||
| otal for VKORC | 3673 (-1639 | 428 | র ব | 98.06% | 432 | 0 | 100 | 99.889 | ||||||
| ---------------------------------------------------------------------- | Final Result* | I | ||||||||||||
| SamplesTestedGenotype | Tests PerSite | Site | GenotypeCalls | CorrectCalls | ncorrectCalls | CallsNo | % CorrectCalls | ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Confidence | CorrectCalls | ncorrectCalls | CallsNo | CorrectCalls0/0 | 95% One-SideoConfidence | |
| 432 | 429 | 99.3 | ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ | 432 | 100 | Lower Limit 99.88% 99.88% | ||||||||
| Total per Site | ---------------------------------------------------J | 432 | 423 | 97.9 | 96.45% 99.31% | 432 | 100 | |||||||
| ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ | 43210-4-4-4-4-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6-6 | 43 T | ਰੇ 8 | 432 | 100 | 99.88% | ||||||||
| Total for AssaThe Call Company of Children And Children of Children Company of Children of | 1296 | 1283 | 13 | 98.99 | 98.42% | 296 | 100 | 99.96% |
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{7}------------------------------------------------
Results after 10 minutes of samples
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ults after one repeat of samples repeat OK
1: no results - ip sense ligude des bourne of was buffer on one sample; correct test result achieved on secon
.
Second test result: Correct, shown on first page.
Note: No calibration sample was attached.
c Site 3: one sample had no call; correct test result achieved on second test
| Genotype | 2C9 | *2 | *3 | VKORCI3673 (-16390 | #SamplesTested | Samples withGenotype Callsmade byINFINITIa | Samples withCorrectCallsb | Samples with NoCalls | Samples withIncorrectCalls | First Time Run |
|---|---|---|---|---|---|---|---|---|---|---|
| *1/*2 | *1/*3 | AA | 36 | 34 | 34 | 2 | 0 | |||
| *1/*1 | *1/*1 | GG | 36 | 36 | 36 | 0 | 0 | |||
| *2/*2 | *1/*1 | GA | 36 | 35 | 35 | 1 | 0 | |||
| *1/*1 | *3/*3 | GA | 36 | 35 | 35 | 1 | 0 |
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8
{8}------------------------------------------------
Drug Interference
Evaluation of potential interference from bilirubin, cholesterol, and heparin demonstrated that presence of these compounds in concentrations of 8mg/dl bilirubin, 70mg/dl cholesterol and 133v/dl heparin does not interfere with the INFINITI 2C9 & VKORC1 Multiplex Assay.
Sample Carry-Over
No sample carry-over was detected when 300ng of a positive sample was followed by 10ng of a second positive sample, and when 300ng of a positive sample was followed by a "No Template Control" or water. All genotype calls were 100% correct.
Assay Interference
Running the INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin and the INFINITI Assay for Factor II & Factor V on the same instrument did not affect the results of the assays, i.e., the INFINITI 2C9 & VKORC 1 Multiplex Assay for Warfarin did not affect the results of the INFINITI Assay for Factor II & Factor V, and vise versa.
Reagent Stability
| BioFilmChip Microaray: | 12 months at RT (15-30°C) |
|---|---|
| Intellipac Reagent: | 12 months Refrigerated (2-8°C) |
| Amplification Mix: | 12 months Frozen (-10°C) |
Conclusion
The above pre-clinical and clinical data support the safety and effectiveness of the INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin.
{9}------------------------------------------------
Image /page/9/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo features a stylized eagle with its wings spread, and the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" are arranged in a circular pattern around the eagle. The logo is black and white and appears to be a simple, clean design.
Public Health Service
Food and Drug Administration 2098 Gaither Road Rockville MD 20850
JAN 2 8 2008
Autogenomics, Inc Ms. Evelyn Lopez 2251 Rutherford Road Carlsbad, CA 92008
Re: K073014
Trade/Device Name: INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin Regulation Number: 21 CFR 862.3360 Regulation Name: Drug metabolizing enzyme genotyping system Regulatory Class: Class II Product Code: ODW, ODV, NSU Dated: October 23, 2007 Received: October 29, 2007
Dear Ms. Lopez:
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.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).
{10}------------------------------------------------
Page 2 -
This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.
If you desire specific information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (240) 276-0490. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). 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 (240) 276-3150 or at its Internet address at http://www.fda.gov/cdrh/industry/support/index.html.
Sincerely vours.
Jean M. Coopes, M.S., D.V.M.
Jean M. Cooper, M.S., D.V.M. Director Division of Chemistry and Toxicology Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health
Enclosure
{11}------------------------------------------------
Indications for Use
| 510(k) Number (if known): | K073014 |
|---|---|
| Device Name: | INFINITI 2C9 &VKORC1 Multiplex Assay for Warfarin |
| Indications For Use: | The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is an in vitro diagnostic test for the detection and genotyping of the *2 and *3 CYP4502C9 genetic variants and the VKORC1 3673 (-1639) intronic variant in genomic deoxyribonucleic acid (DNA) obtained from EDTA-anticoagulated whole blood samples. The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is a qualitative assay for use in clinical laboratories upon prescription by the attending physician.The INFINITI 2C9 & VKORC1 Multiplex Assay for Warfarin is indicated for use to identify individuals at risk for sensitivity to warfarin. |
| Prescription Use | X | AND/OR | Over-The-Counter Use |
|---|---|---|---|
| (Part 21 CFR 801 Subpart D) | (21 CFR 801 Subpart C) |
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)
Division Sign-Off
Office of In Vitro Diagnostic Device Evaluation and Safety
073016 510-1
Page 1 of ____________________________________________________________________________________________________________________________________________________________________
§ 862.3360 Drug metabolizing enzyme genotyping system.
(a)
Identification. A drug metabolizing enzyme genotyping system is a device intended for use in testing deoxyribonucleic acid (DNA) extracted from clinical samples to identify the presence or absence of human genotypic markers encoding a drug metabolizing enzyme. This device is used as an aid in determining treatment choice and individualizing treatment dose for therapeutics that are metabolized primarily by the specific enzyme about which the system provides genotypic information.(b)
Classification. Class II (special controls). The special control is FDA's guidance document entitled “Class II Special Controls Guidance Document: Drug Metabolizing Enzyme Genotyping Test System.” See § 862.1(d) for the availability of this guidance document.