ESENSOR WARFARIN SENSITIVITY SALIVA TEST

K110786 · GenMark Diagnostics, Inc. · NSU · Dec 2, 2011 · Clinical Chemistry

Device Facts

• Record ID: K110786
• Device Name: ESENSOR WARFARIN SENSITIVITY SALIVA TEST
• Applicant: GenMark Diagnostics, Inc.
• Product Code: NSU · Clinical Chemistry
• Decision Date: Dec 2, 2011
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.2570
• Device Class: Class 2

Intended Use

The eSensor® Warfarin Sensitivity Saliva Test is an in vitro diagnostic test for the detection and genotyping of the *2 and *3 alleles of the cytochrome P450 (CYP450) 2C9 gene locus and the Vitamin K epoxide reductase C1 (VKORC1) gene promoter polymorphism (-1639G>A) from genomic DNA of human saliva samples collected using the Oragene® Dx Device, as an aid in the identification of patients at risk for increased warfarin sensitivity.

Device Story

The eSensor® Warfarin Sensitivity Saliva Test is a multiplex microarray-based genotyping system. It processes genomic DNA extracted from human saliva (collected via Oragene® Dx). The assay uses PCR amplification, exonuclease digestion, and competitive DNA hybridization in a sandwich format. Target DNA binds to sequence-specific solution-phase signal probes and solid-phase electrode-bound capture probes on a cartridge. The eSensor® XT-8 instrument uses alternating current voltammetry (ACV) to detect ferrocene-labeled signal probes bound to the electrodes. The system is operated by laboratory personnel in a clinical setting. The instrument provides automated data analysis and generates a report of the patient's genotype. This information assists physicians in identifying patients at risk for increased warfarin sensitivity, though therapeutic dosing decisions must incorporate additional clinical factors.

Clinical Evidence

No clinical trials; analytical performance only. Reproducibility studies (n=10 and n=14 donors) across multiple sites/operators showed 100% agreement after re-testing. Method comparison study (n=316 samples) against bi-directional sequencing showed 99.05% total assay correct call rate. Interfering substances (endogenous/exogenous) showed no impact on genotyping accuracy.

Technological Characteristics

Multiplex microarray-based genotyping system. Components: eSensor XT-8 instrument and disposable cartridge. Cartridge features: PCB with 72 gold-plated working electrodes, microfluidic diaphragm pump, and EEPROM. Sensing principle: Electrochemical detection via ACV of ferrocene-labeled probes. Connectivity: USB data upload, barcode scanner. Software: Automated analysis of hybridization signals. Sterilization: Not specified.

Indications for Use

Indicated for patients requiring warfarin therapy to identify genetic risk for increased sensitivity. Detects CYP450 2C9 *2 and *3 alleles and VKORC1 (-1639G>A) polymorphism from saliva-derived genomic DNA. For use by licensed laboratories.

Regulatory Classification

Identification

Instrumentation for clinical multiplex test systems is a device intended to measure and sort multiple signals generated by an assay from a clinical sample. This instrumentation is used with a specific assay to measure multiple similar analytes that establish a single indicator to aid in diagnosis. Such instrumentation may be compatible with more than one specific assay. The device includes a signal reader unit, and may also integrate reagent handling, hybridization, washing, dedicated instrument control, and other hardware components, as well as raw data storage mechanisms, data acquisition software, and software to process detected signals.

Special Controls

*Classification.* Class II (special controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9. The special control is FDA's guidance document entitled “Class II Special Controls Guidance Document: Instrumentation for Clinical Multiplex Test Systems.” See § 862.1(d) for the availability of this guidance document.

The special control is FDA's guidance document entitled "Class II Special Controls Guidance Document: Instrumentation for Clinical Multiplex Test Systems."

Predicate Devices

• eSensor Warfarin Sensitivity Test (K073720)

Submission Summary (Full Text)

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DEC - 2 2011 K110786

#### 510(k) SUMMARY: eSensor® Warfarin Sensitivity Saliva Test on XT-8 System

Preparation Date: October 24, 2011

Submitted By: Clinical Micro Sensor dba GenMark 5964 La Place Court, Suite100 Carlsbad, CA 92008 USA Phone: [phone] Fax: [phone]

#### Contacts:

John Riolo, Vice President - Quality & Regulatory Affairs (Official Correspondent)

#### Proprietary Names and Classifications:

For the assay: eSensor® Warfarin Sensitivity Saliva Test (Kit)

Regulation: 21CFR 864.7280 Panel: Hematology (81) Classification: Class II Product Codes: CYTOCHROME P450 2C9 (CYP450 2C9) DRUG METABOLIZING ENZYME GENOTYPING SYSTEM (ODW) and VITAMIN K EPOXIDE REDUCTASE COMPLEX SUBUNIT ONE (VKORC1) GENOTYPING SYSTEM (ODV)

For the instrument: eSensor® XT-8 Instrument (System) Regulation: 21CFR 862.2570 Panel: Clinical Chemistry (75) Classification: Class II Product Code: NSU - Instrument for Clinical Multiplex Test Systems

#### Common name:

Warfarin Sensitivity Saliva CYP450 2C9 *2 and CYP450 2C9 *3 alleles and VKORC1 Genotyping

#### Intended use:

The eSensor® Warfarin Sensitivity Saliva Test is an in vitro diagnostic test for the detection and genotyping of the *2 and *3 alleles of the cytochrome P450 (CYP450) 2C9 gene locus and the Vitamin K epoxide reductase C1 (VKORC1) gene promoter polymorphism (-1639G>A) from genomic DNA of human saliva samples collected using the Oragene® Dx Device, as an aid in the identification of patients at risk for increased warfarin sensitivity.

#### Special conditions for use statement(s):

The eSensor® Warfarin Sensitivity Saliva Test can provide information to the physician and lab director on the genotype and allele designation of the three (3) polymorphisms stated in the intended use. However, determination of the therapeutic strategy and treatment dose for Warfarin will depend on multiple additional factors and are beyond the scope of the eSensor® Warfarin Sensitivity

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Saliva Test Report and Package Insert.

The eSensor® Warfarin Saliva Sensitivity Test is for in vitro diagnostic use within a licensed laboratory, as defined by the Clinical Laboratory Improvement Amendments (CLIA) of 1988.

#### Predicate devices:

K073720 eSensor Warfarin Sensitivity Test eSensor XT-8 System

### Device Description:

The eSensor® Warfarin Sensitivity Saliva Test is a multiplex microarray-based genotyping test system. It is based on the principles of competitive DNA hybridization using a sandwich assay format, wherein a single-stranded target binds concurrently to sequence-specific solution-phase signal probe and solid-phase electrode-bound capture probe. The test employs polymerase chain reaction amplification, exonuclease digestion and hybridization of target DNA. In the process, the double stranded PCR amplicons are digested with exonuclease to generate single stranded DNA suitable for hybridization. Hybridization occurs in the eSensor XT-8 Cartridge (described below) where the singlestranded target DNA is mixed with a hybridization solution containing labeled signal probes.

During hybridization, the target DNA binds to a complementary, single-stranded capture probe immobilized on the working electrode surface. Single-stranded signal probes (labeled with electrochemically active ferrocenes) bind to the target adjacent to the capture probe. When inserted into the eSensor XT-8 instrument (described below), simultaneous hybridization of target to signal probes and capture probe is detected by alternating current voltammetry (ACV). Each pair of working electrodes on the array contains a different capture probe, and sequential analysis of each electrode allows genotyping of multiple mutations or polymorphisms.

## The Assay Cartridge (eSensor XT-8 Cartridge)

The eSensor XT-8 cartridge device consists of a printed circuit board (PCB) with a multi-layer laminate and a plastic cover that forms a hybridization chamber has a volume of approximately 140 µl. The cartridge consists of a diaphragm pump and check valves (microfluidic components) that circulate the hybridization solution in the hybridization chamber when inserted into the eSensor XT-8 instrument. The PCB chip consists of an array of 72 gold-plated working electrodes, a silver/silver chloride reference electrode, and two gold-plated auxiliary electrodes. Each working electrode has a connector contact pad on the opposite side of the chip for electrical connection to the eSensor XT-8 instrument. Each electrode is modified with a multicomponent, self-assembled monolayer that includes presvnthesized oligonucleotide capture probes specific for each polymorphic site on the test panel and insulator molecules. The cartridge also contains an electrically erasable programmable read-only memory component (EEPROM) that stores information related to the cartridge (e.g., assay identifier, cartridge lot number, and expiration date).

## The eSensor XT-8 Instrument (Same as cleared under k073720)

The eSensor XT-8 is a clinical multiplex instrument that has a modular design consisting of a base module and one, two, or three cartridge-processing towers containing 8, 16, or 24 cartridge slots, respectively. The cartridge slots operate independently of each other. Any number of cartridges can be loaded at one time, and the remaining slots are available for use while the instrument is running.

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The base module controls each processing tower, provides power, and stores and analyzes data. The base module includes the user interface, and a 15-in. portrait-orientation display and touch panel. The instrument is designed to be operated solely with the touch screen interface. Entering patient accession numbers and reagent lot codes can be performed by the bar code scanner, the touch screen, or uploading a text file from a USB memory stick.

Each processing tower consists of eight cartridge modules, each containing a cartridge connector, a precision-controlled heater, an air pump, and electronics. The air pumps drive the diaphragm pump and valve system in the cartridge, eliminating fluid contact between the instrument and the cartridge. The pneumatic pumping enables recirculation of the hybridization solution allowing the target DNA and the signal probes to hybridize with the complementary capture probes on the electrodes. The diaphragm pump in the cartridge is connected to a pneumatic source from the eSensor XT-8 instrument and provides unidirectional pumping of the hybridization mixture through the microfluidic channel during hybridization. Using microfluidic technology to circulate the hybridization solution minimizes the unstirred boundary layer at the electrode surface and continuously replenishes the volume above the electrode that has been depleted of complementary targets and signal probes.

The XT-8 instrument provides electrochemical detection of bound signal probes by ACV and subsequent data analysis and test report generating functions. All hybridization, ACV scanning and analysis parameters are defined by a scanning protocol loaded into the XT-8 Software, and then specified for use by the EEPROM on each cartridge.

#### The Assay Kit

The Warfarin Sensitivity Saliva Test consists of the test cartridge and the following components: 1) PCR REAGENTS consisting of: PCR Mix [PCR buffer containing primers and dNTP mixture (dCTP, dGTP, dATP, and dUTP)], MgCl2 thermostable DNA polymerase (Taq Polymerase ); and 2 GENOTYPING REAGENTS consisting of: lambda exonuclease, signal probes and hybridization buffer ingredients (Buffer-1 and Buffer-2).

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## Comparison to technological features of the predicate devices:

.

The following is a comparison of the GenMark Diagnostics eSensor® Warfarin Sensitivity Saliva Test on the XT-8 System to the predicate: Warfarin Sensitivity Test (K073720)

| Characteristic | eSensor® Warfarin Sensitivity Test<br>(Predicate: K073720) | eSensor® Warfarin Sensitivity Saliva Test |
|---------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------|
| Test type | Qualitative genetic test for single nucleotide<br>polymorphism detection | Same as predicate |
| Sample Type | Genomic DNA obtained from a human whole blood<br>sample | Genomic DNA<br>obtained from a<br>human saliva sample |
| Target of<br>detection | Single-nucleotide polymorphism | Same as predicate |
| DNA extraction | Performed off-line | Same as predicate |
| Genes | CYP450 2C9 gene *2 allele<br>CYP450 2C9 gene *3 allele<br>VKORC1 gene -1639G>A polymorphism | Same as predicate |
| Number of Loci<br>genotyped | 3 | Same as predicate |
| Genotyping<br>reaction location | Test cartridge | Same as predicate |
| Genotyping<br>principle | Sandwich hybridization test | Same as predicate |
| Instrument<br>operating system | eSensor® Instrument Model XT-8 Random access<br>compatible with multiple simultaneous test types. | Same as predicate |
| Assay results | Assay signal results are interpreted by a software<br>program and are assigned a result that is presented<br>to the end-user in a report format | Same as predicate |

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## Performance Characteristics:

## Method Comparison

In a method comparison study, a total of 316 gDNA samples extracted from saliva specimen with A260-280 ratios of 1.2-2.1 were genotyped using the eSensor® Warfarin Sensitivity Saliva Test and DNA sequencing.

r

| Before Retest | | | |
|--------------------------|-----------|-----------|-----------|
| DNA Sequencing<br>Result | 2C9 wt/wt | 2C9 wt/*2 | 2C9*2/*2 |
| Correct Calls | 234 | 69 | 9 |
| No-Calls | 3 | 0 | 0 |
| Miscalls¹ | 0 | 1 | 0 |
| %Agreement | 98.7% | 98.6% | 100.0% |
| 95% LCB | 96.8% | 93.4% | 71.7% |
| DNA Sequencing<br>Result | 2C9 wt/wt | 2C9 wt/*3 | 2C9 *3/*3 |
| Correct Calls | 274 | 34 | 3 |
| No-Calls | 3 | 0 | 0 |
| Miscalls² | 2 | 0 | 0 |
| %Agreement | 98.2% | 100.0% | 100.0% |
| 95% LCB | 96.3% | 91.6% | 36.8% |
| DNA Sequencing<br>Result | VKORC1 | VKORC1 | VKORC1 |
| Correct Calls | G/G | G/A | A/A |
| Correct Calls | 120 | 131 | 62 |
| No-Calls | 2 | 1 | 0 |
| Miscalls | 0 | 0 | 0 |
| %Agreement | 98.4% | 99.2% | 100.0% |
| 95% LCB | 94.9% | 96.5% | 95.3% |

,

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| eSensor® Warfarin Sensitivity Saliva Test Method Comparison<br>After Retest | | | |
|-----------------------------------------------------------------------------|-----------|-----------|-----------|
| DNA Sequencing<br>Result | 2C9 wt/wt | 2C9 wt/*2 | 2C9*2/*2 |
| Correct Calls | 237 | 69 | 9 |
| No-Calls | 0 | 0 | 0 |
| Miscalls1 | 0 | 1 | 0 |
| %Agreement | 100.0% | 98.6% | 100.0% |
| 95% LCB | 98.7% | 93.4% | 71.7% |
| DNA Sequencing<br>Result | 2C9 wt/wt | 2C9 wt/*3 | 2C9 *3/*3 |
| Correct Calls | 277 | 34 | 3 |
| No-Calls | 0 | 0 | 0 |
| Miscalls2 | 2 | 0 | 0 |
| %Agreement | 99.3% | 100.0% | 100.0% |
| 95% LCB | 97.8% | 91.6% | 36.8% |
| DNA Sequencing<br>Result | VKORC1 | VKORC1 | VKORC1 |
| Result | G/G | G/A | A/A |
| Correct Calls | 122 | 132 | 62 |
| No-Calls | 0 | 0 | 0 |
| Miscalls | 0 | 0 | 0 |
| %Agreement | 100.0% | 100.0% | 100.0% |
| 95% LCB | 97.6% | 97.8% | 95.3% |

#### Interference Substances

Effect of Endogenous Interfering Substances: Interfering substances including salivary a-amylase, hemoglobin, immunoglobulin A (IgA) and total protein were spiked into saliva samples at the highest amounts found in literature. 10 donors provided five saliva samples each spiked with one of the four interfering substances. A control sample was included. Three extractions were performed on each sample. There was 100% agreement between the eSensor® Warfarin Sensitivity Saliva Test results and bidirectional DNA sequencing for all test substances in first pass, demonstrating no effect of any interfering substances on genotyping.

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| Substance | Concentration | Samples Tested | Correct Calls | Incorrect Calls | No-Calls | % Agreement |
|---------------|------------------|----------------|---------------|-----------------|----------|-------------|
| Control | NA | 30 | 30 | 0 | 0 | 100% |
| Amylase | 260 ± 45 U/mL | 30 | 30 | 0 | 0 | 100% |
| Hemoglobin | 20 mg/mL | 30 | 30 | 0 | 0 | 100% |
| IgA | 188 ± 80 mg/L | 30 | 30 | 0 | 0 | 100% |
| Total Protein | 1.46 ± 0.4 mg/mL | 30 | 30 | 0 | 0 | 100% |

Effect of Exogenous Interfering Substances: Potentially interfering exogenous substances (eating, drinking, chewing gum, using mouthwash and smoking) introduced into saliva samples through various activities were tested. Each activity group was composed of five donors who each provided samples for baseline, immediate and 30 minutes. Three samples per donor were tested. There was 100% agreement between the eSensor® Warfarin Sensitivity Saliva Test results and bidirectional DNA sequencing for all activities tested in first pass, demonstrating no effect of any interfering substances on genotyping.

| Activity | Time-point | Samples<br>Tested | Correct Calls | Incorrect<br>Calls | No-Calls | %<br>Agreement |
|-------------|------------|-------------------|---------------|--------------------|----------|----------------|
| Eating | Baseline | 15 | 15 | 0 | 0 | 100% |
| Eating | Immediate | 15 | 15 | 0 | 0 | 100% |
| Eating | 30 minutes | 15 | 15 | 0 | 0 | 100% |
| Drinking | Baseline | 15 | 15 | 0 | 0 | 100% |
| Drinking | Immediate | 15 | 15 | 0 | 0 | 100% |
| Drinking | 30 minutes | 15 | 15 | 0 | 0 | 100% |
| Chewing Gum | Baseline | 15 | 15 | 0 | 0 | 100% |
| Chewing Gum | Immediate | 15 | 15 | 0 | 0 | 100% |
| Chewing Gum | 30 minutes | 15 | 15 | 0 | 0 | 100% |
| Mouthwash | Baseline | 15 | 15 | 0 | 0 | 100% |
| Mouthwash | Immediate | 15 | 15 | 0 | 0 | 100% |
| Mouthwash | 30 minutes | 15 | 15 | 0 | 0 | 100% |
| Smoking | Baseline | 15 | 15 | 0 | 0 | 100% |
| Smoking | Immediate | 15 | 15 | 0 | 0 | 100% |
| Smoking | 30 minutes | 15 | 15 | 0 | 0 | 100% |

## Conclusion:

The above clinical test results support the safety and effectiveness of the eSensor® Warfarin Sensitivity Saliva Test on the eSensor® XT-8 System, and demonstrate substantial equivalence to the predicate device.

eSensor® is a registered trademark of CMS dba GenMark and its subsidiaries

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Image /page/7/Picture/1 description: The image shows the logo for the Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" around the perimeter. Inside the seal is an abstract symbol resembling a stylized human figure or bird.

10903 New Hampshire Avenue Silver Spring, MD 20993

GenMark Diagnostics, Inc. c/o John Riolo 5964 La Place Court, Suite 100 Carlsbad, CA 92008

DEC - 2 2011

Re: k110786 Trade Name: eSensor Warfarin Sensitivity Saliva Test Regulation Number: 21 CFR §862.3360 Regulation Name: Drug Metabolizing Enzyme Genotyping System Regulatory Class: Class II Product Codes: ODW, ODV, NSU Dated: October 24, 2011 Received: October 27, 2011

Dear Mr. Riolo:

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); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

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If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding postmarket surveillance, please contact CDRH's Office of Surveillance and Biometric's (OSB's) Division of Postmarket Surveillance at [phone]. For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/Medical

Devices/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 [phone] or [phone] or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm

Sincerely yours.

N

Counney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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# Indications for Use Form

510(k) Number (if known): K110786

Device Name: eSensor® Warfarin Sensitivity Saliva Test

Indications for Use:

The eSensor® Warfarin Sensitivity Saliva Test is an in vitro diagnostic for the detection and genotyping of the *2 and *3 alleles of the cytochrome P450 (CYP450) 2C9 gene locus and the Vitamin K epoxide reductase C1 ( VKORC1) gene promoter polymorphism (-1639G>A) from qenomic DNA extracted from human saliva samples collected using the the Oragene® Dx Device, as an aid in the identification of patients at risk for increased warfarin sensitivity.

> Prescription Use _____________________________________________________________________________________________________________________________________________________________ (21 CFR 801 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE OF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

Signature

Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety

510(k): K110786

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