Method Comparison Study of the Spartan FRX CYP2C19 Genotyping System Against Bi-directional Sequencing

The Spartan FRX CYP2C19 Test System (hereafter referred to as the 'FRX System') is a qualitative in vitro diagnostic test for the identification of a patient's CYP2C19 *2, *3 and *17 genotypes from genomic DNA obtained from buccal swab samples.

The FRX system is comprised of hardware and consumable components. The hardware components of the system include an Analyzer (thermal cycler with fluorescence detection capability), a netbook computer and a printer. The consumable component of the FRX system is a sample collection kit. Each kit contains a buccal swab (used to collect the patient sample) and a tube containing the reagents required for genomic DNA extraction and PCR amplification stages of the test.

The Spartan FRX System is capable of detecting three CYP2C19 SNPs (*2, *3, *17) in each test performed. An individual sample collection kit is required for each SNP tested; therefore three sample collection kits are required for each test performed on the system.

The FRX System is to be used by trained personnel in CLIA certified laboratories and is for use with buccal samples collected directly from patients. The FRX CYP2C19 test is intended to enable clinicians to identify patients with mutations in the *2,*3 and *17 loci of the CYP2C19 gene and is indicated for use as an aid to clinicians in determining strategies for therapeutics that are metabolized by the CYP2C19 gene product.

To perform a test, the user collects three buccal samples from the patient and then inserts a sample into each of the three reagent tubes (one for each of the CYP2C19 loci *2, *3 and *17). The reagent tubes are placed into the Analyzer and the FRX system automates the processes of DNA extraction, PCR amplification, fluorescent signal detection and data analysis. The system provides the user with a printed result listing the patient genotypes at the *2, *3 and *17 loci.

This study has been designed to demonstrate the concordance (positive and negative percent agreement) by comparing the Spartan FRX system against bi-directional DNA sequencing. The study will also evaluate the concordance between three different sample transport embodiments provided to laboratories for use in transporting the reagent tubes with collected buccal sample swab to the laboratory.

For each mutation, agreement of the FRX System against bidirectional sequencing (comparative method) will be calculated: The overall accuracy of the test will be calculated and reported as follows:

• of concordant results between the test and the reference method Accuracy = ----------------------------------------------------------------
• of samples analyzed by the reference method

The first and second pass results from the study will be analyzed and tabulated as outlined in the table below. The acceptance criteria for the overall study will be percent in agreement ≥ 99.0% for the second pass with the lower bound of a one-sided 95% confidence interval using the score method ≥ 95.0%.