INFINITI CYP2C19 ASSAY

K101683 · Autogenomics, Incorporated · NTI · Oct 25, 2010 · Clinical Toxicology

Device Facts

• Record ID: K101683
• Device Name: INFINITI CYP2C19 ASSAY
• Applicant: Autogenomics, Incorporated
• Product Code: NTI · Clinical Toxicology
• Decision Date: Oct 25, 2010
• Decision: SESE
• Submission Type: Abbreviated
• Regulation: 21 CFR 862.3360
• Device Class: Class 2

Indications for Use

The INFINITI CYP2C19 Assay is an in vitro diagnostic test for the identification of a patient's CYP450 2C19 genotype in genomic deoxyribonucleic acid (DNA) obtained from EDTA-anticoagulated whole blood samples. The INFINITI CYP2C19 Assay is a qualitative assay for use in clinical laboratories upon prescription by the attending physician.

Device Story

The INFINITI CYP2C19 Assay is an in vitro diagnostic system for genotyping CYP450 2C19 (*2, *3, *17) from EDTA-anticoagulated whole blood. The process involves DNA extraction, PCR amplification, allele-specific primer extension, and hybridization to a BioFilmChip Microarray. The INFINITI Analyzer automates sample handling, reagent management, hybridization, and signal detection via a built-in confocal microscope. The system uses an Intellipac Reagent Module with an integrated memory chip for reagent tracking. Results are presented as genotype calls to assist clinicians in therapeutic strategy. The device is intended for use in clinical laboratories by trained personnel. It provides qualitative genetic information to guide clinical decision-making regarding drug metabolism, though it is not intended to predict specific drug response or non-response.

Clinical Evidence

Bench testing only. Performance evaluated via analytical specificity, limit of detection (LOD), and reproducibility studies. LOD established at 20ng DNA/test. Comparison to bi-directional sequencing (n=317) showed 100% agreement after repeat testing of 'no call' samples. Inter-laboratory reproducibility (n=685 total tests) demonstrated a 96.9% correct call rate across three sites, four operators, and four instruments.

Technological Characteristics

Microarray-based genotyping system. Components: BioFilmChip Microarray (polyester film with multi-layer components), Intellipac Reagent Module, and PCR Amplification Mix. Sensing principle: Signature Tag/Capture probe hybridization under isothermal conditions with confocal microscopy detection. Connectivity: Standalone analyzer. Software: Automated signal detection and genotype calling.

Indications for Use

Indicated for use as an aid to clinicians in determining therapeutic strategy for therapeutics metabolized by the CYP450 2C19 gene product, specifically *2, *3, and *17 alleles. Not indicated to predict drug response or non-response.

Regulatory Classification

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.

Special Controls

The special control is FDA's guidance document entitled "Class II Special Controls Guidance Document: Drug Metabolizing Enzyme Genotyping System."

*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.

Predicate Devices

• INFINITI Warfarin Assay (K073014)
• AmpliChip CYP450 Microarray (K043576)

Submission Summary (Full Text)

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# 510(K) SUMMARY

| Submitted By | AutoGenomics, Inc.<br>2980 Scott Street<br>Vista, CA 92081, USA<br>Telephone: [phone]<br>FAX: [phone]<br><br>Contact: Evelyn Lopez<br>Vice President, Regulatory Affairs<br><br>Date Prepared: October 22, 2010 | K101683<br>OCT 25 2010 |
|----------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------|
| Device Name | Trade or Proprietary Name: | INFINITI CYP2C19 Assay |
| | Common or Usual Name: | Cytochrome P450 CYP2C19 Test |
| Regulations and<br>Product Codes | Regulations: | 21CFR§862.3360 Drug Metabolizing Enzyme Genotyping Systems<br>21CFR§862.2570 Instrumentation for Clinical Multiplex Test Systems |
| | Classification: | Class II |
| | Product Codes: | NTI Drug Metabolizing Enzyme Genotyping System<br>NSU Instrumentation for Clinical Multiplex Test Systems |
| Predicate Device | (a) INFINITI Warfarin Assay (k073014)<br>(b) AmpliChip CYP450 Microarray (k043576) | |
| Device Description | The INFINITI CYP2C19 Assay is an <i>in vitro</i> diagnostic device which utilizes proprietary<br>film-based microarray technology combined with process automation, reagent<br>management, and software technology for the detection and genotyping of the 2C19 *2,<br>*3, and *17 mutations in genomic deoxyribonucleic acid (DNA) obtained from EDTA-<br>anticoagulated whole blood samples. | |
| | The INFINITI CYP2C19 Assay is comprised of the BioFilmChipTM Microarray, the<br>Intellipac Reagent Module and the PCR Amplification Mix. The INFINITI CYP2C19<br>Assay should be run using the AutoGenomics INFINITI Analyzer. | |
| | The BioFilmChip Microarray consists of a polyester film coated with proprietary multi-<br>layer components designed for DNA analysis. The layers have been designed to provide<br>a versatile surface to enhance test performance. There can be up to 240 spots per<br>microarray with each spot representing a different allele. The microarrays are designed to<br>be assay specific. | |
| | The Intellipac Reagent Module contains up four reservoirs that house the test reagents and<br>has an integrated memory chip. Information on the reagent such as lot number, expiration<br>date and remaining tests, are archived in the memory. | |
| | The PCR Amplification Mix consists of the reagents needed for the PCR amplification<br>step of the assay. | |

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The INFINITI CYP2C19 Assay 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 Steps (c) through (f) are automated by the INFINITI Analyzer. The INFINITI Analyzer automates the 2C19 assay and integrates all the discrete processes of sample (PCR amplicon) handling, reagent management, hybridization, and results The assays are processed automatically and read by the built-in confocal analysis. microscope. Results are analyzed and presented as genotype calls. Intended Use The INFINITI CYP2C19 Assay is an in vitro diagnostic test for the identification of a patient's CYP450 2C19 genotype in genomic deoxyribonucleic acid (DNA) obtained from EDTA-anticoagulated whole blood samples. The INFINITI CYP2C19 Assay is a qualitative assay for use in clinical laboratories upon prescription by the attending physician. Indication for Use The INFINITI CYP2C19 Assay is indicated for use as an aid to clinicians in determining therapeutic strategy for therapeutics that are metabolized by the CYP450 2C19 gene product, specifically *2, *3, *17. The INFINITI CYP2C19 Assay is not indicated to be used to predict drug response or nonresponse. Substantial Table 1a and Table 1b provide a comparison of the INFINITI CYP2C19 Assay to the Equivalence predicate devices. The comparison demonstrates that the INFINITI CYP2C19 Assay is substantially equivalent to the predicate devices.

| | Predicate | Subject Device |
|---------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------|
| Characteristics | INFINITI Warfarin Assay<br>(k073014) | INFINITI CYP2C19 Assay |
| | 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 EDTA-anticoagulated whole blood sample | Same |
| Reaction Conditions | Utilizes thermal cycling<br>Utilizes target DNA amplification<br>Reactions occur on a single biofilm microarray chip | Same |
| Assay Results | Assay signal results are interpreted by a software program<br>Assay results are provided as genotype calls reported to the end user in a report format | Same |
| Instrumentation | INFINITI Analyzer | Same |
| | Differences | |
| Target Genes | CYP4502C9 and VKORC1 | CYP4502C19 |

- Table ta

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| Characteristics | Predicate | Subject Device |
|--------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | AmpliChip CYP450 Microarray (k043576) | INFINITI CYP2C19 Assay |
| | <i>Similarities</i> | |
| Indication for use | As an aid in determining therapeutic strategy<br>and treatment dose for therapeutics that are<br>metabolized by the CYP450 2C19 gene<br>product. | As an aid in determining therapeutic strategy<br>for therapeutics that are metabolized by the<br>CYP450 2C19 gene product. Not intended to<br>predict drug response or non-response |
| DNA sequence | Detects specific DNA sequences through<br>recognition of DNA targets | Same |
| Specimen Type | Purified DNA from EDTA-anticoagulated<br>whole blood sample | Same |
| Technology | Microarray-based genotyping test for<br>simultaneous detection (multiplex system) of<br>DNA sequences | Same |
| Reaction<br>Conditions | Utilizes thermal cycling Utilizes target DNA amplification 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 |
| Target Gene | CYP450 2C19 | Same |
| | <i>Differences</i> | |
| Target Gene<br>Mutations | CYP450 2C19 *2 and *3 | CYP450 2C19 *2, *3, and *17 |

Performance The following are performance characteristics of the INFINITI CYP2C19 Assay:

## 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)

The analytical sensitivity (Limit of Detection) of the INFINITI CYP2C19 Assay was assessed by analysis of whole blood samples at serial dilutions representing 500ng, 100ng, 100ng, 50ng, 20ng, 10ng and 5ng DNA input (per test) to determine the lowest level of genomic DNA (ng input per test) that would give a ≥ 90% correct call rate of the allele with no incorrect calls.

The following whole blood samples were used in the LOD studies: * 1/*1, * 1/*17, * 1/*17, *2/*2 and *2/*17. Sample genotype was determined by bi-directional sequencing.

Two extraction methods were used in the studies: Qiagen Q1Aamp DNA Blood Kit and the PSS Magtration System. The concentration and A260/A280 were determined by spectrophotometry for the extracted DNA.

A total of 1,560 tests were completed for the LOD studies. A ≥ 90% correct call rate with no incorrect calls for the allele was obtained at DNA input levels from 400ng/test to 5ng/test. There was one incorrect call at the 5ng DNA input level. The incorrect call was probably due to the low DNA concentration, therefore, we are not recommending DNA input level at this low concentration.

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The lowest detectable level for the INFINITI CYP2C19 Assay is 20ng DNA per test. This is less than one-half (1/2) the recommended DNA input level of 50ng/test, and four times the level at which an incorrect call was made by the assay. We believe that establishing the lowest detectable level at 200g DNA/test is conservative and should by and an incorrect call.

Table 2 provides a summary of the LOD studies for the INFINITI CYP2C19 Assay.

| Genotype4 | | ngDNA : } | | | | | % correct » | :95% One-sideo |
|-----------|----------------|----------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | Sample | input per | Replicates | | Correct Incorrect | | . calls 15 | Confidence |
| | ID | test | tested · | calls | calls | | time.run | Lower Limit |
| | | | માં, તુ<br>1. 1.4 2.4 | | 11:21:34 : 1 | | - الوقوب الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع الموقع المو | : ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ ・ |
| | | 200 | 40 | 34 | 0 | б | 85.0% | 72.7% |
| | AG44 | 400 | 40 | 38 | 0 | 2 | 95.0% | 87.0% |
| | AG105 | 200 | 40 | 38 | 0 | 2 | 95.0% | 87.0% |
| | AG208 | 100 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| * 1/* I | AG209 | રેપ | 40 | ਤੇ ਰੇ | 0 | l | 97.5% | 91.4% |
| | AG211 | 20 | 40 | 39 | 0 | l | 97.5% | 91.4% |
| | AG219 | 10 | 40 | 38 | 0 | 2 | 95.0% | 87.0% |
| | | 5 | 40 | 39 | 0 | 1 | 97.5% | 91.4% |
| | Total | | ~320<br>: | 305<br>. . | 0 - | । ਤ<br>t | 95.3% | - 92.8% - - 92.8% - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - |
| | | 500 | 20 | . ਉ | 0 | l | 95.0% | 82.9% |
| | | 400 | 20 | 20 | 0 | 0 | 100% | 97.5% |
| | AG81 | 200 | 20 | 20 | 0 | 0 | 100% | 97.5% |
| | AG197 | 100 | 20 | 20 | 0 | 0 | 100% | 97.5% |
| *1/*2 | AG210<br>AG223 | રુ | 40 | 37 | 0 | 3 | 92.5% | 83.1% |
| | | 20 | 40 | 36 | 0 | 4 | 90.0% | 79.5% |
| | | 10 | 40 | 38 | 0 | 2 | 95.0% | 87.0% |
| | | 5 | 40 | ਤੇ ਰੇ | 0 | 1 | 97.5% | 91.4% |
| | Total | | 240" | : 229 : | 0 - 0 - 1 - | 11 | 95.4%<br>ﺮ ﺍﻟﻤﺮﺍﺟﻊ<br>【】 | 92.6% |
| | AG82 | 200 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | | 400 | 40 | 39 | 0 | l | 97.5% | 91.4% |
| | | 200 | 40 | 37 | 0 | 3 | 92.5% | 83.1% |
| | AG199 | 100 | 40 | 39 | 0 | l | 97.5% | 91.4% |
| *2/*2 | AG214 | રી | 40 | 37 | 0 | 3 | 92.5% | 83.1% |
| | AG233 | 20 | 40 | 38 | 0 | 2 | 95.0% | 87.0% |
| | | 10 | 40 | 37 | 0 | 3 | 92.5% | 83.1% |
| | | 5 | 40 | 37 | 0 | 3 | 92.5% | 83.1% |
| | Total<br>r ¿ , | | - 320 | 4 . 304 | 200000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000 | . 16 | 95.0% - 2 | 92.5% |
| *1/*3 | | 500 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | | 400 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | | 200 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | AG152<br>AG162 | 100 | 40 | 39 | 0 | l | 97.5% | 91.4% |
| | | રુભ | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | | 20 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | | 10 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | | 5 | 40 | 38 | 1 | 1 | 95.0% | 87.0% |
| | Total | | 320 : --<br>、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、、 | 317 - 317 - | 1945 1 1 2 2 2 2 2 2 | | | 199.1% = 1 + 97.9% |
| Genotype | Sample ID | ngDNA input per test | Replicates tested | Correct calls | Incorrect calls | No calls | % correct calls 1st time run | 95% One-sided Confidence Lower Limit |
| *1/*17 | AG74 | 500 | 40 | 36 | 0 | 4 | 90.0% | 79.5% |
| | AG94 | 400 | 40 | 38 | 0 | 2 | 95.0% | 87.0% |
| | AG180 | 200 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | AG235 | 100 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | | 50 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | | 20 | 40 | 38 | 0 | 2 | 95.0% | 87.0% |
| | | 10 | 40 | 40 | 0 | 0 | 100% | 98.8% |
| | | 5 | 40 | 39 | 0 | 1 | 97.5% | 91.4% |
| Total | | 320 | 311 | 0 | 9 | 97.2% | 95.2% | Total |
| *2/*17 | AG191 | 100 | 20 | 20 | 0 | 0 | 100% | 97.5% |
| | AG195 | 5 | 20 | 20 | 0 | 0 | 100% | 97.5% |
| Total | | | 40 | 40 | 0 | 0 | 100% | 98.8% |

#### Table 2 Limit of Detection

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a determined by bi-directional sequencing

### Percent Agreement vs. Bi-directional Sequencing

The INFINITI CYP2C19 Assay 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 CYP2C19 Assay. Patient samples were de-identified to protect patient's identity.

- . A total of 317 samples were tested.
- . There were no incorrect calls
- . Six samples (1.9%) had to be repeated because of "no call" due to NTCE is reported when the quality or quantity of the DNA in the sample/PCR product is poor. All six no calls gave correct calls upon the repeat for a 100% agreement of the INFINITI CYP2C19 Assay with bi-directional sequencing. The repeat test was done on the same extracted DNA. Only one repeat was done for each sample.

The results of the comparison studies comparing the INFINITI CYP2C19 Assay to bi-directional sequencing are provided in Table 3.

| Genotype | Number Tested | Replicates per Sample | Number of Correct Genotype Calls | Number of Incorrect Calls | No Calls | Agreement | 95% One-sided Confidence Lower Limit |
|----------|---------------|-----------------------|----------------------------------|---------------------------|----------|-----------|--------------------------------------|
| *1/*1 | 105 | 1 | 103 | 0 | 2 | 98.1% | 95.0% |
| *1/*2 | 80 | 1 | 77 | 0 | 3 | 96.2% | 91.5% |
| *2/*2 | 12 | 1 | 12 | 0 | 0 | 100% | 95.8% |
| *1/*3 | 8 | 1 | 8 | 0 | 0 | 100% | 93.8% |
| *3/*3 | 1 | 1 | 1 | 0 | 0 | 100% | 50.0% |
| *1/*17 | 74 | 1 | 73 | 0 | 1 | 98.6% | 95.3% |
| *17/*17 | 16 | 1 | 16 | 0 | 0 | 100% | 96.9% |
| *2/*3 | 4 | 1 | 4 | 0 | 0 | 100% | 87.5% |
| *2/*17 | 16 | 1 | 16 | 0 | 0 | 100% | 96.9% |
| *3/*17 | 1 | 1 | 1 | 0 | 0 | 100% | 50.0% |
| Total | 317 | 1 | 311 | 0 | 6 | 98.1% | 96.4% |

Table 3 Agreement between INFINITI CYP2C19 Assay with Bi-directional Sequencing

a Genotype determined by bi-directional sequencing; *1/*1 samples are wild-type for *2, *3 and *17

#### Assay Inter-Laboratory Reproducibility

A three-site study was conducted to demonstrate the reproducibility of the INFINITI CYP2C19 Assay. The study involved three identical lots of the INFINITI CYP2C19 Assay, four operators, and four instruments (one site ran two sets of reproducibility studies, each with a different operator and instrument).

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The sites ran identical samples comprised of 12 whole blood samples. The sites were blinded to sample identity. At each site, each sample was run in duplicate per day/operator for five non-consecutive days. A total of 430 tests were run. Of the 430 samples assayed, 14 samples (3.3%) had to be repeated due to "No Calls". The samples were repeated using the same extracted DNA. These NTCE errors might have been caused by temperature gradient during PCPc.imrooner sealing of the PCR tubes, or operator pipetting error. The genotype calls from the repeat assays were 100% correct, There was one incorrect call (*1/*2 instead of *2/*2). The root cause of the incorrect call was not definitively determined.

Results of the inter-laboratory reproducibility study are summarized in Table 4a and Table 4b.

| Genotype* | Samples<br>Tested | Siteb | Replicates<br>per Site | Replicates with Genotype<br>Calls made by<br>INFINITI | Correct<br>Calls | Incorrect<br>Callsd | No<br>Callse | % Correct<br>Callsf | 95% One-sided<br>Confidence -<br>Lower Limitg |
|-----------|-------------------|-------|------------------------|-------------------------------------------------------|------------------|---------------------|--------------|---------------------|-----------------------------------------------|
| *1/*1 | 2 | 1 | 40 | 39 | 39 | 0 | 1 | 97.5 | 91.4 |
| | | 2 | 20 | 20 | 20 | 0 | 0 | 100 | 97.5 |
| | | 3 | 10 | 10 | 10 | 0 | 0 | 100 | 95.0 |
| | | total | 70 | 69 | 69 | 0 | 1 | 98.6 | 95.1 |
| *1/*2 | 3 | 1h | 40 | 40 | 40 | 0 | 0 | 100 | 98.8 |
| | | 2 | 30 | 30 | 30 | 0 | 0 | 100 | 98.3 |
| | | 3 | 30 | 30 | 30 | 0 | 0 | 100 | 98.3 |
| | | total | 100 | 100 | 100 | 0 | 0 | 100 | 99.5 |
| *2/*2 | 2 | 1 | 40 | 39 | 38 | 1 | 1 | 95.0 | 87.0 |
| | | 2 | 20 | 20 | 20 | 0 | 0 | 100 | 97.5 |
| | | 3 | 20 | 14 | 14 | 0 | 6 | 70.0 | 47.4 |
| | | total | 80 | 73 | 72 | 1 | 7 | 90.0 | 82.8 |
| *1/*3 | 1 | 1 | 20 | 20 | 20 | 0 | 0 | 100 | 97.5 |
| | | 2 | 10 | 10 | 10 | 0 | 0 | 100 | 95.0 |
| | | 3 | 10 | 10 | 10 | 0 | 0 | 100 | 95.0 |
| | | total | 40 | 40 | 40 | 0 | 0 | 100 | 98.8 |
| *1/*17 | 2 | 1 | 40 | 39 | 39 | 0 | 1 | 97.5 | 91.4 |
| | | 2 | 20 | 17 | 17 | 0 | 3 | 85.0 | 66.9 |
| | | 3 | 10 | 9 | 9 | 0 | 1 | 90.0 | 66.4 |
| | | total | 70 | 65 | 65 | 0 | 5 | 92.9 | 86.1 |
| *17/*17 | 2 | 1 | 40 | 40 | 40 | 0 | 0 | 100 | 98.8 |
| | | 2 | 20 | 19 | 19 | 0 | 1 | 95.0 | 82.9 |
| | | 3 | 10 | 10 | 10 | 0 | 0 | 100 | 95.0 |
| | | total | 70 | 69 | 69 | 0 | 1 | 98.6 | 95.1 |
| Total | 12 | All | 430 | 416 | 415 | 1d | 14e | 96.5 | 94.7 |

Table 4a Inter-Laboratory Reproducibility of the INFINITI CYP2C19 Assay by Genotype Call

determined by bi-directional sequencing; * 1/* 1 samples are wild-type for *2, * 3 and * 17

ь Internal site (Site 1) had two sets, one operator each

ﺑ Excludes samples with No Calls

o Initial result was incorrect (*1/*2 instead of *2/*2). The root cause of the incorrect call was not definitively determined.

One no call was due to Registration Spot Error - this error is reported when the microarray chip is not properly aligned. Repear test gave the correct call

13 reported NTCE Error - NTCE is reported if the quality of the DNA in the sample/PCR product is poor. Repeat tests gave the correct calls

{ Samples with correct calls/samples tested

8 Site 3: one sample had A260/A280 of 1.16 (1.6 is required), therefore only one sample was tested

h Site 1: one sample had A260/A280 of 1.46 (1.6 is required), therefore only two samples were tested

.. -Site 3: one sample had A260/A280 of 1.46 (1.6 is required), therefore only one sample was tested

J Site 3: one sample had A260/A280 of 1.45 (1.6 is required), therefore only one sample was tested

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| Table 4b | Inter-Laboratory Reproducibility of the INFINITI CYP2C19 Assay by Sample | | | | | | | |
|-----------|--------------------------------------------------------------------------|-----------------------|-------------------------------------------------|----------------------------|-----------------------|------------------------------|-----------------------|--------------------------------------|
| Sample ID | Genotype | Replicates per sample | Replicates with Genotype Calls made by INFINITI | Samples with Correct Calls | Samples with No Calls | Samples with Incorrect Calls | Correct Call Rate (%) | 95% One-sided Confidence Lower Limit |
| 1 | *2/*2 | 40 | 38 | 37 | 2 | 1 | 92.5 | 83.1 |
| 2 | *2/*2 | 40 | 35 | 35 | 5 | 0 | 87.5 | 76.0 |
| 3 | *17/*17 | 40 | 39 | 39 | 1 | 0 | 97.5 | 91.4 |
| 4 | *1/*17 | 40 | 37 | 37 | 3 | 0 | 92.5 | 83.1 |
| 5 | *1/*3 | 40 | 40 | 40 | 0 | 0 | 100 | 98.8 |
| 6 | *1/*2 | 40 | 40 | 40 | 0 | 0 | 100 | 98.8 |
| 7 | *1/*2 | 40 | 40 | 40 | 0 | 0 | 100 | 98.8 |
| 8 | *1/*1 | 30 | 29 | 29 | 1 | 0 | 96.7 | 88.6 |
| 9 | *17/*17 | 30 | 29 | 29 | 1 | 0 | 96.7 | 88.6 |
| 10 | *1/*1 | 40 | 40 | 40 | 0 | 0 | 100 | 98.8 |
| 11 | *1/*17 | 30 | 29 | 29 | 1 | 0 | 96.7 | 88.6 |
| 12 | *1/*2 | 20 | 20 | 20 | 0 | 0 | 100 | 97.5 |
| | All | 430 | 416 | 415 | 14 | 1 | 96.5 | 94.7 |

Table 4b Inter-Laboratory Reproducibility of the INFINITI CYP2C19 Assay by San

8 Determined by bi-directional sequencing. * 1/*1 samples are wild-type for *2, *3 and *17

b Excludes samples with No Calls

& A sample with correct call indicates a correct call at all loci.

4 One no call was due to Registration Spot Error - this error is reported when the microarray chip is not properly aligned. Repeat test gave the correct call; 13 reported NTCE is reported if the quality or quantity of the DNA in the sample/PCR product is poor. Repeat tests gave the correct calls

6 Initial result was incorrect (*1/*2 instead of *2/*2). The root cause of the incorrect call was not definitively determined

{ Samples with correct calls/samples tested

Additional reproducibil…