The Spartan RX CYP2C19 System is a qualitative in vitro diagnostic test for the identification of a patient's CYP2C19 *2, *3 and *17 genotype determined from genomic DNA obtained from a buccal swab sample. For prescription use only.

Spartan RX CYP2C19 Assay - The Spartan RX CYP2C19 System is indicated for use as an aid to clinicians in determining therapeutic strategies for therapeutics that are metabolized by the Cytochrome P450 2C19 gene product, and that are specifically affected by the *2, *3, and *17 alleles. The Spartan RX CYP2C19 Assay will be run on the Spartan RX CYP2C19 Platform from the buccal sample collected with a buccal swab. The Spartan RX CYP2C19 Assay is not indicated to be used to predict drug response or non-response.

Spartan RX CYP2C19 Platform - The Spartan RX CYP2C19 System is indicated for use as an aid to clinicians in determining therapeutic strategies for therapeutics that are metabolized by the Cytochrome P450 2C19 gene product, and that are specifically affected by the *2, *3, and *17 alleles. The Spartan RX CYP2C19 Platform will be used to run the Spartan RX CYP2C19 Assay.

Device Description

The Spartan RX CYP2C19 System is a sample-to-result DNA testing system that uses proprietary technology to integrate DNA extraction and amplification. Genotypes are determined using PCR and fluorescent probe detection. The Spartan RX CYP2C19 System is comprised of the Spartan RX liadrooom prable a and Spartan RX CYP2C19 Assays. The Spartan RX CYP2C19 Assays are run on the Spartan RX CYP2C19 Platform.

The Spartan RX CYP2C19 System is based on the following processes:

Buccal swab collection i.
ii. DNA extraction
PCR-based amplification of the target gene lii.
Detection of the *2, *3, and *17 alleles using fluorescent-probes iv.
Fluorescent signal detection and analysis V.
Genotype determination vi.

The Spartan RX CYP2C19 System integrates and automates steps ii to vi. Results are presented to the end user as genotype calls. The system also has integrated controls that monitor performance of a run and automatically inform the user of any anomalies in the instrument or reagents.

The system detects the CYP2C19 *2, *3, and *17 genotypes in separate reagent tubes. The operator collects buccal swab samples from a patient; inserts each sample into a reagent tube; and then inserts the reagent tubes into a Spartan RX Analyzer instrument.

AI/ML Overview

The Spartan RX CYP2C19 System is a qualitative in vitro diagnostic test for the identification of a patient's CYP2C19 *2, *3, and *17 genotype from buccal swab samples. The system comprises the Spartan RX Platform (instrumentation) and Spartan RX CYP2C19 Assays (consumables). It integrates DNA extraction, PCR-based amplification, fluorescent probe detection, and genotype determination.

Acceptance Criteria and Reported Device Performance

Study Parameter	Acceptance Criteria (Implicit)	Reported Device Performance (First Pass)	Reported Device Performance (Second Pass)
Limit of Detection (LOD)	High percentage of correct calls, especially with typical or low DNA input.	84.6% (5 pooled swabs), 100.0% (2 pooled swabs), 98.1% (Normal Swab), 98.1% (1 Half Stroke), 84.6% (Inside Mouth Touch)	98.1% (5 pooled swabs), 100.0% (2 pooled swabs), 100.0% (Normal Swab), 100.0% (1 Half Stroke), 100.0% (Inside Mouth Touch)
Method Comparison	High percentage of agreement with bi-directional sequencing.	98.8% correct call rate (overall)	100.0% correct call rate (overall)
Inter-Laboratory Reproducibility	High percentage of correct calls across different sites and operators.	98.9% correct call rate (overall)	99.8% correct call rate (overall)
Reagent Lot-to-Lot Reproducibility	Consistent performance across different reagent lots.	Lot 1: 97% correct, Lot 2: 100% correct, Lot 3: 100% correct	Lot 1: 100% correct, Lot 2: 100% correct, Lot 3: 100% correct
Exogenous and Endogenous Interfering Substances	High percentage of correct calls in presence of common interfering substances.	91.5% correct call rate (overall), with some variation depending on substance (e.g., toothpaste 31.3%)	99.55% correct call rate (overall)

Note: The acceptance criteria are implicitly inferred from the reported performance results, which consistently show very high percentages of correct calls and agreement. A lower limit of detection study indicates an acceptable first pass correct call rate of 84.6% at input levels lower than 0.1 swabs per test.

Study Details

Sample sizes used for the test set and data provenance:
- Limit of Detection (LOD):
  - Part 1: 100 individual buccal swabs collected from 40 different individuals.
  - Part 2: 52 samples were tested per condition (5 conditions, total 260 tests), collected from *1/*1, *2/*17, *17/*17, and *2/*3 individuals.
  - Data Provenance: Not explicitly stated, but implies clinical samples possibly from Canada where the submitter is located. Retrospective or prospective is not explicitly mentioned for sample collection, but the samples were analyzed as part of a validation study.
- Method Comparison: 327 samples tested, with 325 included in the analysis.
  - Data Provenance: Not explicitly stated, but implies clinical samples. Samples were de-identified.
- Inter-Laboratory Reproducibility: 960 tests performed (8 individuals * 2 operators * 2 sessions * 5 days * 3 sites).
  - Data Provenance: Not explicitly stated, but implies clinical samples or samples with confirmed genotypes.
- Reagent Lot-to-Lot Reproducibility: 107 samples (Lot 1), 106 samples (Lot 2), 107 samples (Lot 3) in the first pass testing.
  - Data Provenance: Not explicitly stated.
- Exogenous and Endogenous Interfering Substances: 16 samples tested for each of 14 substances (total 224 tests).
  - Data Provenance: Buccal swab samples collected from *1/*1, *2/*17, *17/*17, and *2/*3 individuals where genotypes were confirmed.
Number of experts used to establish the ground truth for the test set and their qualifications:
- The ground truth for all studies was established by bi-directional sequencing. This is a molecular biology technique, not a human expert interpretation. Geneticists or molecular biologists would be involved in performing and interpreting the sequencing results, but specific numbers and qualifications are not mentioned.
Adjudication method for the test set:
- The study design employed a "second pass" re-test for samples that initially yielded "No calls." For the Method Comparison and Inter-Laboratory Reproducibility studies, if a first pass resulted in a "No call," the sample was re-tested. This serves as an internal adjudication or re-evaluation mechanism. No explicit "2+1" or "3+1" expert adjudication is described, as the ground truth is objective sequencing data.
Multi-reader multi-case (MRMC) comparative effectiveness study:
- No MRMC comparative effectiveness study was done. The device is a diagnostic test that provides a genotype call, not an imaging device or aid to human interpretation that would typically involve human readers.
Standalone (i.e., algorithm only without human-in-the-loop performance) study:
- Yes, the performance studies (Limit of Detection, Method Comparison, Inter-Laboratory Reproducibility, Reagent Lot-to-Lot Reproducibility, Exogenous and Endogenous Interfering Substances) represent standalone performance of the Spartan RX CYP2C19 System. The system integrates and automates the DNA testing process, and results are presented as genotype calls directly, indicating an algorithm-only performance assessment against sequencing-derived ground truth.
Type of ground truth used:
- Bi-directional sequencing was used as the ground truth for all performance studies. This is a highly accurate and widely accepted method for confirming DNA sequences and, thus, genotypes.
Sample size for the training set:
- The document does not explicitly mention a separate training set or its sample size. The reported studies primarily describe validation/test set performance. For molecular diagnostic devices, the development often involves internal optimization and algorithm tuning using various samples, but these are not typically referred to as a "training set" in the same way as machine learning models.
How the ground truth for the training set was established:
- Since a distinct "training set" is not explicitly defined or discussed in the document for performance evaluation, the method for establishing its ground truth is also not detailed. However, it can be inferred that any samples used during the development and optimization phases would also likely have been characterized using highly accurate methods like bi-directional sequencing, similar to the test sets.

Summary

{0}------------------------------------------------

510(k) Summary

Submitter Name:	Spartan Bioscience Inc.
Address:	Suite 204, 6 Gurdwara RdOttawa, Ontario, Canada K2E 8A31 (613) 228-7756
Contact:	Dr. Chris HarderVP Diagnostics
Phone:	1 (613) 228-7756
E-mail:	chris.harder@spartanbio.com
Date Prepared:	Aug 9th, 2013
Device Trade Name:	Spartan RX TM CYP2C19 SystemFor the assay: Spartan RX CYP2C19 AssayFor the platform: Spartan RX CYP2C19 Platform
Device Common Name:	Drug metabolizing enzyme genotyping assay
Measurand:	CYP450 2C19 2, 3, *17
Sample Type:	DNA from buccal cells
Technology:	Polymerase Chain Reaction (PCR)
Device Panel:	Chemistry and Toxicology
Classification Name:	Drug metabolizing enzyme genotyping system, 862.3360Instrumentation for clinical multiplex test systems, 862.2570
Classification Code:	NTI: Drug metabolizing enzyme genotyping systemNSU: Instrumentation for clinical multiplex test systems
Predicate Device:	INFINITI CYP2C19 Assay, 510(k) Number K101683Classification Code: NTI and NSU, Regulation No. 862.3360 and862.2570

Intended Use

Indication for Use

{1}------------------------------------------------

a buccal swab. The Spartan RX CYP2C19 Assay is not indicated to be used to predict drug response or non-response.

Device Description

The Spartan RX CYP2C19 System is based on the following processes:

Buccal swab collection i.
ii. DNA extraction
PCR-based amplification of the target gene lii.
Detection of the *2, *3, and *17 alleles using fluorescent-probes iv.
Fluorescent signal detection and analysis V.
Genotype determination vi.

Spartan RX CYP2C19 Platform

There are four components of the Spartan RX CYP2C19 Platform.

a) The SPARTAN RX ANALYZER is a thermal cycling instrument that automates polymerase chain reaction (PCR) amplification of the target gene, fluorescence-based detection of CYP2C19 alleles, and genotype calling.
The NETBOOK serves as the user interface for logging in to the Spartan RX Platform, inputting b) patient information, and starting a test.
The PRINTER automatically prints the genotype results after the Spartan RX Platform finishes c) performing the Spartan RX CYP2C19 Assays.
d) The BARCODE SCANNER is used to automatically enter SCK and ECK lot numbers into the Spartan RX Platform.

Spartan RX CYP2C19 Assays

The Spartan RX CYP2C19 Assays are the consumable components of the Spartan RX CYP2C19 System. There are two different types of assay-each type is described in detail below.

a) SAMPLE COLLECTION KITS (SCKs) contain the consumables required to determine a patient's CYP2C19*2, *3, or *17 genotype. There are three types of SCKs, specific for the *2, *3, or *17 alleles. Each SCK consists of the following components:

{2}------------------------------------------------

POUCH, containing labeling information to ensure traceability (lot number, manufacturing date, and expiry date) and two compartments (for the buccal swab and reagent tube). The pouch also includes a barcode that is compatible with a standard reader.
BUCCAL SWAB, used to collect a buccal sample from the inside of a patient's cheek and transfer it into the reagent tube.
REAGENT TUBE, containing chemicals for DNA extraction, PCR amplification, and fluorescent detection of the specific CYP2C19 allele.

The SCKs are color-coded to indicate which CYP2C19 allele they are designed to detect.

BLUE = *2 י
WHITE = *3 ・
BLACK = *17
b) EXTERNAL CONTROL KITS (ECKs) contain the consumables required to determine if the Spartan RX Platform and reagents are performing correctly. Each ECK consists of the following components:
- POUCH, containing labeling information to ensure traceability (lot number, manufacturing date, and expiry date).
- REAGENT TUBE, containing synthetic nucleic acid control and chemicals for PCR amplification and fluorescent detection of the specific CYP2C19 allele.

The ECKs are color-coded to indicate which reagent they control for.

BLUE = *2 ・
- WHITE = *3
- BLACK = *17

The ECKs have been modified so that the tube cannot be opened. They are meant to be put directly into the analyzer without adding or removing anything from them

Spartan RX Accessories

The SAMPLE TRANSPORT SYSTEM is an insulated transport bag that contains a polystyrene box and a Cold Block. The Transport System is designed to keep the SCKs cold as they are moved from the freezer to the patient and from the patient to the Spartan RX Analyzer. The Transport System is provided with purchase of the platform and is to be used for all tests..

Substantial Equivalence Discussion

The Spartan RX CYP2C19 System uses the same fundamental scientific technologies, and has a similar intended use, as that of the predicate device-the INFINITI CYP2C19 Assay. The table below provides a comparison of the Spartan RX CYP2C19 System and the predicate device. The comparison demonstrates that the Spartan RX CYP2C19 System is substantially equivalent to the predicate device.

Table 1 – Comparison of the Spartan RX CYP2C19 System to the predicate device

Characteristic	Predicate DeviceINFINITI CYP2C19 Assay(K101683)	Subject DeviceSpartan RX CYP2C19 System
	Similarities
Intended Use	The INFINITI CYP2C19 Assay is an invitro diagnostic test for theidentification of apatient's CYP450 2C19 genotype ingenomic deoxyribonucleic acid (DNA)obtained fromEDTA-anticoagulated whole bloodsamples. The INFINITI CYP2CI9	The Spartan RX CYP2C19 System is aqualitative in vitro diagnostic test for theidentification of a patient's CYP2C19 2,3 and *17 genotype determined fromgenomic DNA obtained from a buccalswab sample. For prescription use only.
Characteristic	Predicate DeviceINFINITI CYP2C19 Assay(K101683)	Subject DeviceSpartan RX CYP2C19 System
	Assay is a qualitativeassay for use in clinical laboratoriesupon prescription by the attendingphysician.
Indication for Use	The INFINITI CYP2C19 Assay isindicated for use as an aid to cliniciansin determiningtherapeutic strategy for therapeuticsthat are metabolized by the CYP4502C19 gene product,specifically 2, 3, * 17.The INFINITI CYP2C19 Assay is notindicated to be used to predict drugresponse or nonresponse.	Spartan RX CYP2C19 Assay - TheSpartan RX CYP2C19 System isindicated for use as an aid to clinicians indetermining therapeutic strategies fortherapeutics that are metabolized by theCytochrome P450 2C19 gene product,and that are specifically affected by the2, 3, and 17 alleles. The Spartan RXCYP2C19 Assay will be run on theSpartan RX CYP2C19 Platform fromthe buccal sample collected with abuccal swab. The Spartan RXCYP2C19 Assay is not indicated to beused to predict drug response or non-response.Spartan RX CYP2C19 Platform - TheSpartan RX CYP2C19 System isindicated for use as an aid to clinicians indetermining therapeutic strategies fortherapeutics that are metabolized by theCytochrome P450 2C19 gene product,and that are specifically affected by the2, 3, and 17 alleles. The Spartan RXCYP2C19 Platform will be used to runthe Spartan RX CYP2C19 Assay.
Limitations	Not intended to be used to predict drugresponse or non-response.	Same
DNA Sequence	Detects specific DNA sequencesthrough recognition of DNA targetswith fluorescence.	Same
Technology	Utilizes thermal cycling and targetDNA amplification.	Same
Assay Results	Assay signal results are interpreted bya software program.Assay results are provided asgenotype calls reported to the enduser in a report format.	Same
Target Gene	CYP4502C19 2, 3, and *17.	Same
		Differences
Specimen Type	Purified DNA from EDTA-anti-coagulated whole blood sample.	Unpurified DNA from a buccal swabsample.
Platform	Microarray-based genotyping test forsimultaneous (multiplex system) ofDNA sequences.	Fluorescent-probe PCR-basedgenotyping test for multiplex analysis ofDNA sequences.
		Trial Calling Rates
Reproducibility (%agreement/ 95%Lower CI)	96.9% / 95.6%	99.8% / 99.4%
Method comparison(% agreement/ 95%Lower CI)	100.0% / 99.8%	100.0% / 99.2%

{3}------------------------------------------------

ﻓ

{4}------------------------------------------------

Performance

Limit of Detection (Analytical Sensitivity)

The input material for the Spartan RX CYP2C19 System is a buccal swab collected from an individual, r his inserted directly into the reagent tube without any requirement for the user to extract or purify White for this reason, the limit of detection (LOD) study was to determine if the Spartan DIV. For the Peacer, In Super of target material amounts collected in typical buccal swabs. The study was conducted in two parts.

Part 1 involved the determination of a relative measure of the amount of DNA present in 100 individual buccal swabs, collected from 40 different individuals by 11 different operators. These operators had laboratory - and non-laboratory backgrounds. The mean DNA amount was deemed to be equivalent to one "typical" swab and all measurements were normalized to this value and presented as the number of swab equivalents.

Part 2 assessed the analytical sensitivity (limit of detection) by analyzing buccal swab samples collected from *1, *2/*17, *17/*17, and *2/*3 individuals. Five DNA collection conditions were used: 5 pooled huccal swabs; 2 pooled buccal swabs; a normal buccal swab; a single downward stroke (1 half stroke); butter wabs, if police based to the inside of the mouth. Replicates of 52 samples were performed for and condition. Results are summarized in Tables 2a and 2b. In parallel, the amount of DNA in the swabs was quantified using SYBR Green (standard curve), and this value was translated to the corresponding number of swabs relative to the mean of the population determined in Part 1.

TestCondition	Genotypea	#Samplestested	#Correctcallsb	# No calls	%Correctcalls	95% One-sidedconfidencelower limit)
5 pooledswabs	1/1	13	11	2	84.6%	63%
	17/17	13	12	1	92.3%	72%
	2/3	13	8	5	61.5%	40%
	2/17	13	13	0	100.0%	83%
	Total	52	44	8	84.6%	75%
2 pooledswabs	1/1	13	13	0	100.0%	83%
	17/17	13	13	0	100.0%	83%
	2/3	13	13	0	100.0%	83%
	2/17	13	13	0	100.0%	83%
	Total	52	52	0	100.0%	95%
Normal Swab	1/1	13	13	0	100.0%	83%
	17/17	13	13	0	100.0%	83%
	2/3	13	12	1	92.3%	72%
	2/17	13	13	0	100.0%	83%
	Total	52	51	1	98.1%	92%
1 Half Stroke	1/1	13	12	1	92.3%	72%
	17/17	13	13	0	100.0%	83%
	2/3	13	13	0	100.0%	83%
	2/17	13	13	0	100.0%	83%
	Total	52	51	1	98.1%	92%
Inside MouthTouch	1/1	13	10	3	76.9%	54%
	17/17	13	12	1	92.3%	72%
	2/3	13	9	4	69.2%	46%
	2/17	13	13	0	100.0%	83%
	Total	52	44	8	84.6%	75%

Table 2a - Limit of Detection - FIRST PASS

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

" uniteryped by br arcolibrar ocquently of all, results of all three genotypes (*2, *3, *17) had to be correct.

in three for a sample to be doomied a cerrest can increased rate of No calls and Internal Control Errors.

{5}------------------------------------------------

TestCondition	Genotypea	#Samplestested	#Correctcallsb	# No calls	%Correctcalls	95% One-sidedconfidencelower limit)
5 pooledswabs	1/1	13	12	1	92.3%	72%
	17/17	13	13	0	100.0%	83%
	2/3	13	13	0	100.0%	83%
	2/17	13	13	0	100.0%	83%
Total		52	51	1	98.1%	92%
2 pooledswabs	1/1	13	13	0	100.0%	83%
	17/17	13	13	0	100.0%	83%
	2/3	13	13	0	100.0%	83%
	2/17	13	13	0	100.0%	83%
Total		52	52	0	100.0%	95%
Normal Swab	1/1	13	13	0	100.0%	83%
	17/17	13	13	0	100.0%	83%
	2/3	13	13	0	100.0%	83%
	2/17	13	13	0	100.0%	83%
Total		52	52	0	100.0%	95%
1 Half Stroke	1/1	13	13	0	100.0%	83%
	17/17	13	13	0	100.0%	83%
	2/3	13	13	0	100.0%	83%
	2/17	13	13	0	100.0%	83%
Total		52	52	0	100.0%	95%
Inside MouthTouch	1/1	13	13	0	100.0%	83%
	17/17	13	13	0	100.0%	83%
	2/3	13	13	0	100.0%	83%
	2/17	13	13	0	100.0%	83%
Total		52	52	0	100.0%	95%

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

" Genotype by brunes moral soquenting," 1) " bainpies is of all three genotypes (*2, *3, *17) had to be correct.

The first pass correct call rate at input levels lower than 0.1 swabs per test was 84.6%. Therefore, the The first pass correct can face at mput levels lower than b. Towals por too may be has was new teet.

{6}------------------------------------------------

Relationship between # Swabs across population & amount of collected material using various swabbing techniques

Image /page/6/Figure/2 description: This image is a graph that plots the number of swabs in a sample versus the sample number. The y-axis, labeled "NUMBER OF SWABS IN SAMPLE", ranges from 0.0 to 4.0. There are also horizontal lines indicating different levels: 3.3 ng, 1.6 ng, 1.3 ng, 0.6 ng, and 0.1 ng. The graph also includes a legend indicating different types of swabs: 5 Pooled Swabs, 2 Pooled Swabs, Normal Swab, One Half Stroke, and Inside Mouth Touch.

Figure 1 – Relationship between the normal swab population & the amount of material collected using the various swabbing techniques as outlined in the protocol. The corresponding average amount of genomic DNA (ng) collected using each swabbing technique is indicated on the right side of the graph.

These experiments indicate that there are two consequences when excessively high or low amounts of buccal material are collected. Either the system generates an internal control abort (Positive System Control fail) or the system generates a No call result. To avoid these consequences, the Instructions for Ountrol half of the applied is swab firmly with 3x up-and-down strokes. Based on these results, proper swabbing is an important factor that influences the rate of No call results. Also, the results from Part 3 demonstrated the system's lower limit of detection when using genomic DNA. The recommended quantity of DNA for external controls was based on these results.

Method Comparison

The Spartan RX CYP2C19 System was compared to bi-directional sequencing. Samples were deidentified to protect patient identity. Genomic DNA was extracted from saliva samples. If the result of the first test for a particular individual was a No call, the test was repeated (second pass).

The Method Comparison study was performed by 10 operators at three different locations. The operators had no previous swabbing experience. Of the 327 samples tested, sequencing was unsuccessful for 2 samples these samples were excluded from the analysis. Of the 325 samples included in the analysis, 4 samples gave a No call result on the first pass test and were re-tested. After the second pass, all samples were called correctly. No samples were called incorrectly. The overall correct call rate for the first pass and second pass tests was 98.8% and 100%, respectively.

For this study, 1/3 of samples collected were tested immediately (4 minutes pre-swabbing and 8 minutes post-swabbing); 1/3 were tested after 10 minutes (4 minutes pre-swabbing and 10 minutes postswabbing); and 1/3 were tested after 60 minutes (45 minutes pre-swabbing and 60 minutes post swabbing) of storage in the Sample Transport System after swab insertion.

Results of first and second pass tests are summarized in Tables 3a and 3b, respectively.

{7}------------------------------------------------

Genotypea	#Samplestested	#CorrectCallsb	# No calls	%Correctcalls	95% One-sidedconfidencelower limit
1/1	130	128	2	98.5%	95%
1/2	95	94	1	98.9%	95%
2/2	19	19	0	100.0%	88%
1/3	7	7	0	100.0%	72%
3/3	1	1	0	100.0%	27%
1/17	40	39	1	97.5%	90%
17/17	11	11	0	100.0%	80%
2/3	6	6	0	100.0%	69%
2/17	15	15	0	100.0%	85%
3/17	1	1	0	100.0%	27%
TOTAL	325	321	4	98.8%	97%

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

" Gellowye by ordinestional soquenting" "1" " oaily of all three genotypes ("2, "3, *17) had to be correct.

Table 3b – Method Comparison – SECOND PASS

Genotypea	#Samplestested	#CorrectCallsb	%Correctcalls	95% One-sidedconfidencelower limit
1/1	130	130	100.0%	98%
1/2	95	95	100.0%	97%
2/2	19	19	100.0%	88%
1/3	7	7	100.0%	72%
3/3	1	1	100.0%	27%
1/17	40	40	100.0%	94%
17/17	11	11	100.0%	80%
2/3	6	6	100.0%	69%
2/17	15	15	100.0%	85%
3/17	1	1	100.0%	27%
TOTAL	325	325	100.0%	99%

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

" Genotype by bronnethonal sequentling, "17" valiptor of all three genotypes (*2, *3, *17) had to be correct.

For the Transport System, the first pass results were 109/110, 105/106,107/109 for the immediate, 10 ron the Transport Oyotom the points, respectively. All results were 100% on the second pass.

{8}------------------------------------------------

Tratel seber Machy & epocal solidition - FIRST PASS

Buccal swab samples were collected from *1/*17, *2/*3, *1/*1, *1/*3, *2/*17, *2/*2, and *1/*2 individuals (one individual per genotype) across three sites. Genotypes of the eight individuals were confirmed by bi-directional sequencing. Two operators were employed at each site, and tests were performed across five non-consecutive days. Typically, operators were laboratory technicians who had no previous experience with swabbing. At one site, two of the operators did not work in the medical or provide oxperience ware two sessions per day (AM and PM), and each session was split into two subsessions. In each sub-session, each of the eight individuals was tested once by Operator 1 and once by Operator 2. If the result of the first test for a particular individual was a No call, the test was repeated (second pass). Results of the inter-laboratory reproducibility study are presented in Tables 4a and 4b.

Sample Type*	Site	# samples tested	# no calls	# Incorrect calls	# correct calls	% correct call rate	95% Confidence Limit
1/1	1	40	0	0	40	100.0%	94%
	3	40	1	0	39	97.5%	90%
	4	40	0	1*	39	97.5%	90%
	Total	120	1	1	118	98.3%	95%
1/2	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	0	0	40	100.0%	94%
	Total	120	0	0	120	100.0%	98%
2/2	1	40	2	0	38	95.0%	86%
	3	40	0	0	40	100.0%	94%
	4	40	0	0	40	100.0%	94%
	Total	120	2	0	118	98.3%	95%
1/3	1	40	1	0	39	97.5%	90%
	3	40	1	0	39	97.5%	90%
	4	40	0	0	40	100.0%	94%
	Total	120	2	0	118	98.3%	95%
2/3	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	3	0	37	92.5%	83%
	Total	120	3	0	117	97.5%	94%
1/17	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	0	0	40	100.0%	94%
	Total	120	0	0	120	100.0%	98%
17/17	1	40	0	0	40	100.0%	94%
	3	40	1	0	39	97.5%	90%
	4	40	1	0	39	97.5%	90%
	Total	120	2	0	118	98.3%	95%
2/17	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	0	0	40	100.0%	94%
	Total	120	0	0	120	100.0%	98%
TOTAL	1	320	3	0	317	99.1%	98%
	3	320	3	0	317	99.1%	98%
	4	320	4	1	315	98.4%	97%
	Total	960	10	1	949	98.9%	98%

Table 4a - Inter-Laboratory Reproducibility - FIRST PASS

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

" Galleryo by a sample to be deemed a correct call, results of all three genotypes (*2, *3, *17) had to be correct.

" The root cause of the incorrect call could not conclusively be determined, but it was not due to bubbles in the reaction tube.

{9}------------------------------------------------

SampleTypea	Site	# samplestested	# nocalls	#incorrectcalls	#correctcallsb	% correct callrate	95%ConfidenceLimit
1/1	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	0	1*	39	97.5%	90%
	Total	120	0	1	119	99.2%	96%
1/2	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	0	0	40	100.0%	94%
	Total	120	0	0	120	100.0%	98%
2/2	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	0	0	40	100.0%	94%
	Total	120	0	0	120	100.0%	98%
1/3	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	0	0	40	100.0%	94%
	Total	120	0	0	120	100.0%	98%
2/3	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	1	0	39	97.5%	90%
	Total	120	1	0	119	99.2%	96%
1/17	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	0	0	40	100.0%	94%
	Total	120	0	0	120	100.0%	98%
17/17	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	0	0	40	100.0%	94%
	Total	120	0	0	120	100.0%	98%
2/17	1	40	0	0	40	100.0%	94%
	3	40	0	0	40	100.0%	94%
	4	40	0	0	40	100.0%	94%
	Total	120	0	0	120	100.0%	98%
TOTAL	1	320	0	0	320	100.0%	99%
	3	320	0	0	320	100.0%	99%
	4	320	1	1	318	99.4%	99%
	Total	960	1	1	958	99.8%	99%

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

Genotype by brunectional bequeneing, " } " call, results of all three genotypes ("2, * 3, * 1 7) had to be correct.

*The rot cause of the incorrect call could not conclusively be determined, but it was not due to bubbles in the reaction tube.

Overall, a total of 960 tests were performed—10 second pass tests were performed due to No calls on the first pass test, and all but 1 were recovered.

Overall, after second pass testing of No call genotype results, the Spartan RX CYP2C19 System Overall, aller Scoonia pass toeing of No San Gerry Jre-sided lower confidence limit of 99%.

{10}------------------------------------------------

Relateetblocklister-llato@ephong.com/lity - SECOND PASS

Lot-to-lot reproducibility was assessed using three independent lots of each reagent type labeled as 1,2 Eot to Toproducibility had account the Site 1 Inter-laboratory Reproducibility study.

On the first pass, 3 samples produced No call results. After the second pass, all of these samples On the first pass, o samples produced the cesults indicate that different reagent lots perform equivalently.

First and second pass results for the Reagent Lot-to-Lot Reproducibility study are presented in Tables 5a and 5b, respectively.

Reagent Lot	Genotype"	# Samplestested	# Correctcallsb	# No calls	% Correct calls(95% one-sidedconfidence lowerlimit)
	2/17	13	13	0	100% (83%)
	1/2	14	14	0	100% (84%)
	1/17	13	13	0	100% (83%)
	17/17	13	13	0	100% (83%)
Lot 1	1/1	14	14	0	100% (84%)
	2/3	13	13	0	100% (83%)
	2/2	13	11	2	85% (63%)
	1/3	14	13	1	93% (73%)
	Total	107	104	3	97% (93%)
	2/17	13	13	0	100% (83%)
	1/2	13	13	0	100% (83%)
	1/17	14	14	0	100% (84%)
	17/17	13	13	0	100% (83%)
Lot 2	1/1	13	13	0	100% (83%)
	2/3	14	14	0	100% (84%)
	2/2	13	13	0	100% (83%)
	1/3	13	13	0	100% (83%)
	Total	106	106	0	100% (98%)
	2/17	14	14	0	100% (84%)
	1/2	13	13	0	100% (83%)
	1/17	13	13	0	100% (83%)
	17/17	14	14	0	100% (84%)
Lot 3	1/1	13	13	0	100% (83%)
	2/3	13	13	0	100% (83%)
	2/2	14	14	0	100% (84%)
	1/3	13	13	0	100% (83%)
	Total	107	107	0	100% (98%)

Table 5a – Reagent Lot-to-Lot Reproducibility – FIRST PASS

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

Genotype by bledited be deemed a correct call, results of all three genotypes (*2, *3, *17) had to be correct.

{11}------------------------------------------------

Reagent Lot	Genotypea	# Samples tested	# Correct calls	% Correct calls (95% one-sided confidence lower limit)
Lot 1	2/17	13	13	100% (83%)
	1/2	14	14	100% (84%)
	1/17	13	13	100% (83%)
	17/17	13	13	100% (83%)
	1/1	14	14	100% (84%)
	2/3	13	13	100% (83%)
	2/2	13	13	100% (83%)
	1/3	14	14	100% (84%)
	Total	107	107	100% (98%)
Lot 2	2/17	13	13	100% (83%)
	1/2	13	13	100% (83%)
	1/17	14	14	100% (84%)
	17/17	13	13	100% (83%)
	1/1	13	13	100% (83%)
	2/3	14	14	100% (84%)
	2/2	13	13	100% (83%)
	1/3	13	13	100% (83%)
	Total	106	106	100% (98%)
Lot 3	2/17	14	14	100% (84%)
	1/2	13	13	100% (83%)
	1/17	13	13	100% (83%)
	17/17	14	14	100% (84%)
	1/1	13	13	100% (83%)
	2/3	13	13	100% (83%)
	2/2	14	14	100% (84%)
	1/3	13	13	100% (83%)
	Total	107	107	100% (98%)

Table 5b – Reagent Lot-to-Lot Reproducibility – SECOND PASS

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

Genotype by bi-directional sequencing; *1,71 samples are wild-type for *2, *3, and *17.
In order for a sample to be deemed a correct call, results of all three genotypes

{12}------------------------------------------------

Exogenous and Endogenous Interfering Substances

Interference from exogenous and endogenous interfering substances was evaluated using buccal swab samples collected from *1/*1, *2/*17, *17/*17, and *2/*3 individuals. Genotypes of all individuals were confirmed by bi-directional sequencing prior to initiation of the study.

To test the impact of exogenous substances, buccal swab samples were collected from individuals after exposure to the substance. The individuals rinsed with water (as per the Instructions for Use for the Spartan RX CYP2C19 System) before swab collection.

Direct exposure of individuals to endogenous substances was not possible. Therefore, these substances were added directly to the reagent tube immediately prior to insertion of the buccal swab sample. Note that the Instructions for Use require the patient to rinse his or her mouth with water prior to collection of the buccal swab sample. The purpose of the water rinse is to remove any contaminating endogenous materials such as blood or blood components. So although certain endogenous substances will interfere with performance of the Spartan RX CYP2C19 System, this risk is mitigated by the water rinse.

The following exogenous and endogenous substances were tested:

Antiseptic mouthwash	–	20 ml - Rinse around mouth for 30 s
Toothpaste	-	3/4" Strip - Brush teeth for 2 min, spit
Baking soda solution	ー	30 ml (0.1 g /ml) – Rinse around mouth for 10 s
Cough syrup	।	30 ml - Rinse around mouth for 10 s
Cranberry juice	-	30 ml – Rinse around mouth for 10 s
Salt water	l	30 ml (0.01 g/ml) - Rinse around mouth for 10 s
Sugar water	-	30 ml (0.01 g/ml) -- Rinse around mouth for 10 s
Meat	–	15 g – Chew for 10 s
Chewing gum	-	1 standard piece - Chew for 1 min
Hard candy	–	1 standard piece - Suck until fully dissolved
Tobacco smoking	-	1 cigar - 5 puffs
Denture paste	I	3 strips – Apply strips to roof of mouth, leave for 5 min,
		remove
Human Oral Bacteria		Approximately 9 x 104 cells (in addition to normal oral flora)
Whole Blood	-	3.5 µl of 0.5% blood (diluted in saliva)

A total of 16 samples were tested for each exogenous and endogenous interfering substance (n=4 per genotype tested). If the result of the first test for a particular individual was a No call, the test was repeated (second pass).

The results of the Exogenous and Endogenous Interfering Substances study indicate that substances commonly introduced into the oral environment do not affect performance of the Spartan RX CYP2C19 System. Nevertheless, proper rinsing and swabbing techniques must be followed to minimize the rate of No calls associated with inhibiting substances.

Results of first and second pass tests performed during the Exogenous and Endogenous Interfering Substances study are summarized in Tables 6a and 6b, respectively.

{13}------------------------------------------------

Genotype	Substance	# Samples tested	# Correct calls	# No calls	% Correct calls	95% one-sided confidence lower limit
Combined genotype results	Mouthwash	16	15	1	93.8%	76%
	Toothpaste	16	5	11	31.3%	16%
	Baking soda	16	14	2	87.5%	68%
	Cough syrup	16	16	0	100.0%	86%
	Cranberry juice	16	16	0	100.0%	86%
	NaCl solution	16	16	0	100.0%	86%
	Sugar solution	16	16	0	100.0%	86%
	Meat (horse)	16	16	0	100.0%	86%
	Chewing gum	16	14	2	87.5%	68%
	Hard candy	16	16	0	100.0%	86%
	Tobacco (cigar)	16	15	1	93.8%	76%
	Denture paste	16	16	0	100.0%	86%
	Bacteria	16	15	1	93.8%	76%
	Whole blood	16	15	1	93.8%	76%
	Total	224	205	19	91.5%	88%

4 In order for a sample to be deemed a correct call, results of all three genotypes (*2, *3, *17) had to be correct.

Table 6b - Exogenous and Endogenous Interfering Substances - SECOND PASS
--------------------------------------------------------------------------	--	--	--

Genotype	Substance	#Samplestested	# Correctcallsa	# Nocalls	% Correctcalls	95% one-sidedconfidencelower limit
	Mouthwash	16	16	0	100.00%	86%
	Toothpaste	16	15	b	93.75%	76%
	Baking soda	16	16	0	100.00%	86%
Combinedgenotyperesults	Cough syrup	16	16	0	100.00%	86%
	Cranberry juice	16	16	0	100.00%	86%
	NaCl solution	16	16	0	100.00%	86%
	Sugar solution	16	16	0	100.00%	86%
	Meat (horse)	16	16	0	100.00%	86%
	Chewing gum	16	16	0	100.00%	86%
	Hard candy	16	16	0	100.00%	86%
	Tobacco (cigar)	16	16	0	100.00%	86%
	Denture paste	16	16	0	100.00%	86%
	Bacteria	16	16	0	100.00%	86%
	Whole blood	16	16	0	100.00%	86%
	Total	224	223	1	99.55%	98%

ª In order for a sample to be deemed a correct call, results of all three genotypes (*2, *3, * 17) had to be correct.

in over for a damps to bo been esignificantly different. For the tist pass, three mointable did not ninse out all of " First and second pass results in tourine and more one all all the took and the tooking the the the neeming the broe the toothpaste before swabing. For the Secure pass, all for the world be and are are are are and and and least three minutes before swabbing.

{14}------------------------------------------------

Image /page/14/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized emblem resembling a caduceus, with three wavy lines emanating from a central point. The emblem is encircled by the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" in a circular arrangement.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

August 12, 2013

Spartan Bioscience Inc. c/o Lorry Huffman c/o Myraqua, Inc. 3 Lagoon Drive, Ste 280 REDWOOD SHORES CA 94065

Re: K123891

Trade/Device Name: Spartan RX™ CYP2C19 System Regulation Number: 21 CFR 862.3360 Regulation Name: Drug metabolizing enzyme genotyping system Regulatory Class: II Product Code: NTI, NSU Dated: June 28, 2013 Received: July 3, 2013

Dear Ms. Huffman:

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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. 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 Part 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

{15}------------------------------------------------

Page 2- Ms. Huffman

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please go to http://www.fda.gov/AboutFDA/CentersOffices/CDRH/CDRHOffices/ucm115809.htm for the Center for Devices and Radiological Health's (CDRH's) Office of Compliance. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/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 (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm.

Sincerely yours,

Carol C. Benson -S for

Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

{16}------------------------------------------------

Indications for Use Form

510(k) Number (if known): _ K123891

Device Name: Spartan RX CYP2C19 System

Indications for Use:

Assay - The Spartan RX CYP2C19 System is indicated for use as an aid to clinicians in determining therapeutic strategies for therapeutics that are metabolized by the Cytochrome P450 2C19 gene product, and that are specifically affected by the *2, *3, and *17 alleles. The Spartan RX CYP2C19 Assay will be run on the Spartan RX Platform from the buccal sample collected with a buccal swab. The Spartan RX CYP2C19 Assay is not indicated to be used to predict drug response or non-response.

Platform - The Spartan RX CYP2C19 System is indicated for use as an aid to clinicians in determining therapeutic strategies for therapeutics that are metabolized by the Cytochrome P450 2C19 gene product, and that are specifically affected by the *2, *3, and *17 alleles. The Spartan RX CYP2C19 Platform will be used to run the Spartan RX CYP2C19 Assay.

Prescription Use	X
(Part 21 CFR 801 Subpart D)

AND/OR

Over-The-Counter 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 Diagnostics and Radiological Health (OIR)

Ruth A. Cheslier -S

Division Sign-Off Office of In Vitro Diagnostics and Radiological Health

K123891 510(K)

Page 1 of 1

Regulation Number and Section

§ 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.