The TruDiagnosis® System is an in vitro diagnostic device intended for processing and genotyping multiple genetic variants in a DNA sample utilizing on-slide PCR gel-drop microarray technology. The TruDiagnosis® System consists of the TruDx® 2000 Imager, the TruArray® Warfarin Sensitivity Test Kit, and the ProFlex™ PCR System using the ProFlex™ 2x Flat Sample Block.

The TruDx® 2000 Imager is an instrument intended for processing and genotyping multiple genetic variants in a DNA sample utilizing on-slide PCR gel-drop microarray technology.

The TruArray® Warfarin Sensitivity Test Kit is an in vitro diagnostic test for the detection and genotyping of the 2C92, 2C93 alleles of the cytochrome P450 (CYP450) 2C9 gene locus and Vitamin K epoxide reductase CL, VKORCI, gene promoter polymorphism (-1639) from genomic DNA of human saliva samples collected using the Oragene® Dx Device (OGD-500) as an aid in the identification of patients at risk for increased warfarin sensitivity. The TruArray® Warfarin Sensitivity Test Kit is a qualitative assay for use in clinical laboratories upon prescription by the attending physician.

Device Description

The TruDiagnosis® System is an in vitro diagnostic device intended for processing and genotyping multiple genes in a DNA sample utilizing on-slide Polymerase Chain Reaction (PCR) gel-drop microarray technology. The TruDiagnosis® System consists of the:

Hardware: TruDx® 2000 Imager ●
Software: TruSpot™ Software ●
. Test Kit: TruArray® Warfarin Sensitivity Test Kit
- TruArray® Test Slide o
- o TruPlex™ reagents
Thermal Cycler: ProFlex™ PCR System using the ProFlex™ 2x Flat Sample Block ●

Hardware: Akonni's microarray imager (TruDx®2000) is an instrument that consists of a highintensity green light emitting diode (LED), custom optics, and a digital grayscale camera.

. The purpose of the TruDx® 2000 Imager is to capture a fluorescence image of the microarray after completing the test.
The user inserts the TruArray® Test Slide into the TruDx® 2000 Imager and follows . the on-screen prompts. The resulting microarray image is automatically analyzed and reported with the TruSpot™ Software.

Software: Akonni's TruSpot™ Software, integrated within the imager, locates and segments each fluorescently labeled microarray Gel-Element and reports signal-to-noise ratios (SNR). Assay results interpreted by TruSpot™ Software program are assigned a genotype and presented to the end user in a report format.

Test Kit: The TruArray® Warfarin Sensitivity Test Kit includes consumables and reagents necessary to perform multiplex on-slide PCR amplification, and fluorogenic target-specific microarray-based hybridization. Specifically:

The TruArray® Test Slide (loaded with target DNA) undergoes thermal cycling via . asymmetric amplification to enrich single stranded fluorescently labeled complimentary strands to capture probes printed on the microarray (asymmetric PCR and allele specific hybridization occur in the same chamber). After hybridization, arrays are washed and dried, and the user then inserts the microarray into the TruDx® 2000 Imager and follows the on-screen prompts.

The warfarin assay performed with the test kit is an in vitro diagnostic test for the detection and genotyping of the 2C92, 2C93 alleles of the cytochrome P450 (CYP450) 2C9 gene locus and Vitamin K epoxied reductase C1, VKORC1 3673, gene promotor polymorphism (-1639) mutations from genomic DNA of human saliva samples collected using the Oragene® Dx OGD-500 Device (K110701).

Thermal Cycler: The ProFlex™ PCR System with the ProFlex™ 2x Flat Sample Block, a component of the TruDiagnosis System, is an end-point thermal cycler, specifically designed for the amplification of nucleic acids using the Polymerase Chain Reaction (PCR) process. The user interface includes a touchscreen with a graphical display that shows the time, status, and temperature for each run. A touchscreen keypad allows you to enter information into fields on the display screen.

AI/ML Overview

Here's an analysis of the acceptance criteria and the studies that prove TruDiagnosis® System meets them:

1. Table of Acceptance Criteria and Reported Device Performance

Based on the provided document, the acceptance criteria are not explicitly stated as distinct pass/fail thresholds in simple terms. Instead, the studies demonstrate the device's performance, aiming for high "Correct Call Rate" and agreement with Sanger sequencing, especially after addressing "No Calls". A reasonable interpretation of the implicit acceptance criteria, based on the studies, would be a high percentage of accurate genotype calls and 100% concordance with the reference method (Sanger sequencing) upon resolution of initial "No Calls".

Performance Characteristic	Acceptance Criteria (Implicit)	Reported Device Performance
Limit of Detection (LoD)	High Correct Call Rate and 100% concordance with Sanger sequencing upon resolution of No Calls down to 2.5 ng DNA input.	First Pass Calls:- 125 ng: 98.4% Correct Call Rate (96.8% LCB)- 25 ng: 98.3% Correct Call Rate (96.7% LCB)- 2.5 ng: 94.4% Correct Call Rate (92.0% LCB)- 0.25 ng: 64.7% Correct Call Rate (60.4% LCB)Final Result (after repeating No Calls):- 125 ng, 25 ng, 2.5 ng: 100.0% Correct Call Rate (99.2% LCB or higher)- 0.25 ng: 63.9% Correct Call Rate (59.6% LCB) - Incorrect calls were not repeated at this concentration.
Method Comparison	100% agreement with bi-directional DNA sequencing.	First Pass Calls (Overall): 98.7% Correct Call Rate (97.0% LCB).Final Result (after repeating No Calls): 99.7% Correct Call Rate (98.4% LCB). "All 3 No Calls were resolved upon the second repeat runs. The Final Results yielded 100% agreement between the TruArray® Warfarin Sensitivity Test Kit results and bi-directional DNA sequencing." (One incorrect call was identified as a sample mix-up, and not re-run).
Endogenous Interference	100% agreement with bi-directional DNA sequencing.	Final Results (after repeating No Calls): 100.0% agreement between the TruArray® Warfarin Sensitivity Test Kit results and bi-directional DNA sequencing for all test substances, demonstrating no effect of any interfering substances on genotyping.
Exogenous Interference	100% agreement with bi-directional DNA sequencing.	Final Results (after repeating No Calls): 100.0% agreement between the TruArray® Warfarin Sensitivity Test Kit results and bi-directional DNA sequencing.
Reproducibility	100% correct call rate across sites, operators, and lots after repeating No Calls.	First Pass Results (Overall): 84.2% (by sample genotype), 85.8% (by SNPs across all loci).Final Results (after repeating No Calls): 100.0% Correct Call Rate across all sample genotypes, sites, operators, and lots (95.1% LCB or higher, 99.2% overall LCB for genotypes, 99.7% overall LCB for SNPs).

2. Sample Sizes and Data Provenance for Test Set

Limit of Detection Study:
- Sample Size: 3 genomic DNA samples (Donors 1, 2, 3), tested at various concentrations with 40 replicates each (total 3 * 4 * 40 = 480 replicates for initial runs, but 362 samples were ultimately tested, implying some initial runs were repeated due to control failures, and some individual test samples were re-run).
- Data Provenance: Genotypes for these 3 donors are provided (*1/*1, *1/*3, AA; *1/*2, *1/*3, AG; *2/*2, *1/*1, GG). The origin of these DNA samples (e.g., country) is not specified. It is a retrospective study since the DNA samples were presumably pre-collected and genotyped by Sanger sequencing.
Method Comparison Study:
- Sample Size: 303 unique human genomic DNA samples.
- Data Provenance: Origin of the data (e.g., country) is not specified, but the samples were isolated from human saliva specimens. It is a retrospective study as the samples were pre-collected using the Oragene® Dx OGD-500 device, extracted, and a reference genotype established.
Interference Studies (Endogenous and Exogenous):
- Endogenous: 5 donors, each providing saliva samples, divided into 5 aliquots (spiked with interferents or un-spiked control). Three extractions performed on each spiked/un-spiked sample. (Total 5 donors * 5 conditions * 3 extractions = 75 samples).
- Exogenous: 5 activity groups (Eating, Drinking, Chewing Gum, Mouth Wash, Smoking) with 5 donors each. Each donor provided 2 saliva samples (baseline and 30 min post-activity), tested in triplicate. (Total 5 groups * 5 donors * 2 timepoints * 3 replicates = 150 samples).
- Data Provenance: Saliva samples from human donors. Origin not specified. Retrospective.
Reproducibility Study:
- Sample Size: Six specific saliva samples (genotypes provided in the table). Each sample was processed in triplicate by 4 operators over 5 non-consecutive days (15 tests per operator per genotype). 6 genotypes * 4 operators * 15 tests = 360 samples.
- Data Provenance: Saliva samples from human donors. Origin not specified. Retrospective for samples, but prospective for the testing process across different sites/operators.

3. Number of Experts and Qualifications for Ground Truth

The document explicitly states that the ground truth for all performance studies (Limit of Detection, Method Comparison, Interference, and Reproducibility) was established by bi-directional Sanger sequencing. It does not mention the use of human experts or their qualifications for establishing this ground truth. Sanger sequencing is a widely accepted gold standard for DNA sequencing and thus acts as the independent reference for genotyping.

4. Adjudication Method for the Test Set

The document describes an adjudication method for "No Calls" found during the initial runs. For "No Calls," the samples were repeated (re-run). Incorrect calls, however, were generally not repeated ("Incorrect Calls were not repeated" in the LoD study footnote, and for the Method Comparison, an incorrect call was attributed to a sample mix-up and "was not re-run"). This indicates an adjudication process where "no-calls" lead to re-testing, but "incorrect calls" (when a result was given but it was wrong) are investigated for root cause and not necessarily re-run for a different result in the study context.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No MRMC comparative effectiveness study was mentioned. The device, the TruDiagnosis® System, is an in vitro diagnostic device for genotyping, which typically involves laboratory testing rather than interpretation by multiple human readers (like radiology images). Therefore, a multi-reader study is not applicable in this context.

6. Standalone Performance

Yes, a standalone performance study was done. The entire performance evaluation (Limit of Detection, Method Comparison, Interference, Reproducibility) assesses the TruDiagnosis® System (which includes the TruArray® Warfarin Sensitivity Test Kit, TruDx® 2000 Imager, and TruSpot™ Software acting together as an algorithm/system) against an independent reference method (bi-directional Sanger sequencing). This is a direct measure of the algorithm's diagnostic accuracy without human intervention in the genotype interpretation after the system generates a result.

7. Type of Ground Truth Used

The ground truth used for all studies was bi-directional Sanger sequencing.

8. Sample Size for the Training Set

The document does not provide information regarding a training set or its sample size. This is typical for a 510(k) submission where the focus is on the validation of the finished device and its performance characteristics against a predicate, rather than the development and training of the underlying algorithm. The "TruSpot™ Software" is mentioned as integrated within the imager and interpreting assay results, implying it's an algorithmic component, but details on its development or training are not included in this summary.

9. How Ground Truth for the Training Set Was Established

Since no information on a training set is provided, there is no detail on how its ground truth was established.

Summary

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA logo on the right. The FDA logo is a blue square with the letters "FDA" in white, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue.

May 24, 2019

Akonni Biosystems Inc. % Rita King, CEO MethodSense, Inc. 1 Copley Parkway, Suite 410 Morrisville, NC 27560

Re: K183530

Trade/Device Name: TruDiagnosis System Regulation Number: 21 CFR 862.3360 Regulation Name: Drug metabolizing enzyme genotyping system Regulatory Class: Class II Product Code: ODW, ODV, NSU Dated: April 26, 2019 Received: May 1, 2019

Dear Rita King:

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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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

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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 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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 comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Kellie B. Kelm, Ph.D. Acting Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K183530

Device Name TruDiagnosis® System

Indications for Use (Describe)

The TruDx® 2000 Imager is an instrument intended for processing and genotyping multiple genetic variants in a DNA sample utilizing on-slide PCR gel-drop microarray technology.

The TruArray® Warfarin Sensitivity Test Kit is an in vitro diagnostic test for the detection and genotyping of the 2C92. 2C93 alleles of the cytochrome P450 (CYP450) 2C9 gene locus and Vitamin K epoxide reductase CL, VKORCI, gene promoter polymorphism (-1639) from genomic DNA of human saliva samples collected using the Oragene® Dx Device (OGD-500) as an aid in the identification of patients at risk for increased warfarin sensitivity. The TruArray® Warfarin Sensitivity Test Kit is a qualitative assay for use in clinical laboratories upon prescription by the attending physician.

Type of Use (Select one or both, as applicable)

Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)

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510(k) Summary

Akonni Biosystems, Inc. (K183530)

This 510(k) Summary is in conformance with 21CFR 807.92

Submitter:	Akonni Biosystems, Inc.400 Sagner AvenueSuite 300Frederick, MD 21701Phone: (301) 698-0101Fax: (301) 698-0202
Primary Contact:	Rita King, CEOMethodSense, Inc.Email: ritaking@methodsense.comPhone: (919) 313-3961Fax: (919) 313-3979
Company Contact:	Charles DaitchCEO
Date Prepared:	May 22, 2019
Device Name and Classification
Trade Name:	TruDiagnosis® System
Common Name:	TruDiagnosis® System
Classification:	Class II
Regulation Number:	21 CFR §862.3360 Drug Metabolizing Enzyme GenotypingSystems21 CFR §864.7750 Prothrombin time test21 CFR §862.2570 Instrumentation for Clinical MultiplexTest Systems
Classification Panel:	Toxicology (91), Hematology (81), Chemistry (75)
Product Code:	ODW- Cytochrom P450 2C9 (CYP450 2C9) Drug MetabolizingEnzyme Genotyping SystemODV- Vitamin K epoxide reductase complex subunit 1 (VKORC1)Genotyping SystemNSU- Instrumentation for Clinical Multiplex Test Systems
Predicate Device:
Predicate Device
Trade Name	eSensor Warfarin Sensitivity Test and XT-8 Instrument
Common Name	eSensor Warfarin Sensitivity Test and XT-8 Instrument
510(k) Submitter / Holder	Osmetech Molecular Diagnostics
510(k) Number	K073720
Regulation Number	21CFR §862.3360 – Drug Metabolism Enzyme GenotypingTest

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	Predicate Device
	21CFR §864.7750 – Prothrombin Time Test21CFR §862.2570 – Instrument for Clinical Multiplex TestSystems
Classification Panel	Toxicology, Hematology, Chemistry
Product Code	ODW Cytochrome P450 2C9 (CYP450 2C9) DrugMetabolizing Enzyme Genotyping System
	ODV Vitamin K epoxide reductase complex subunit 1(VKORC1) Genotyping System
	NSU Instrumentation for Clinical Multiplex Test Systems

Device Description

Hardware: TruDx® 2000 Imager ●
Software: TruSpot™ Software ●
. Test Kit: TruArray® Warfarin Sensitivity Test Kit
- TruArray® Test Slide o
- o TruPlex™ reagents
Thermal Cycler: ProFlex™ PCR System using the ProFlex™ 2x Flat Sample Block ●

Hardware: Akonni's microarray imager (TruDx®2000) is an instrument that consists of a highintensity green light emitting diode (LED), custom optics, and a digital grayscale camera.

. The purpose of the TruDx® 2000 Imager is to capture a fluorescence image of the microarray after completing the test.
The user inserts the TruArray® Test Slide into the TruDx® 2000 Imager and follows . the on-screen prompts. The resulting microarray image is automatically analyzed and reported with the TruSpot™ Software.

The TruArray® Test Slide (loaded with target DNA) undergoes thermal cycling via . asymmetric amplification to enrich single stranded fluorescently labeled complimentary strands to capture probes printed on the microarray (asymmetric PCR and allele specific hybridization occur in the same chamber). After hybridization, arrays are washed and dried, and the user then inserts the microarray into the TruDx® 2000 Imager and follows the on-screen prompts.

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Indications for Use

The TruDx® 2000 Imager is an instrument intended for processing and genotyping multiple genetic variants in a DNA sample utilizing on-slide PCR gel-drop microarray technology.

The TruArray® Warfarin Sensitivity Test Kit is an in vitro diagnostic test for the detection and genotyping of the 2C92, 2C93 alleles of the cytochrome P450 (CYP450) 2C9 gene locus and Vitamin K epoxide reductase Cl, VKORCl, gene promoter polymorphism (-1639) variants from genomic DNA of human saliva samples collected using the Oragene® Dx Device (OGD-500) as an aid in the identification of patients at risk for increased warfarin sensitivity. The TruArray® Warfarin Sensitivity Test Kit is a qualitative assay for use in clinical laboratories upon prescription by the attending physician.

Risk Analysis Method

The TruDiagnosis® System was assessed to determine the reasonably foreseeable sequences/combinations of events leading to hazardous situations and identify and document the harm to the users. A risk analysis was conducted in accordance with ISO 14971:2007 and ISO14971:2012. Medical devices -- Application of risk management to medical devices. Several risks were assessed, including, but not limited to, instrument and assay design, software malfunctions, hardware failure, security, and improper use.

Substantial Equivalence

The table below provides a detailed comparison of the TruDiagnosis System to the predicate device.

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Item	Subject Device	Predicate Device	Comparison
Indications for Use	TruDiagnosis® SystemThe TruDiagnosis® System is an in vitro diagnostic device intended for processing and genotyping multiple genetic variants in a DNA sample utilizing on-slide PCR gel-drop microarray technology. The TruDiagnosis® System consists of the TruDx® 2000 Imager, the TruArray® Warfarin Sensitivity Test Kit, and the ProFlex™ PCR System using the ProFlex™ 2x Flat Sample Block.The TruDx® 2000 Imager is an instrument intended for processing and genotyping multiple genetic variants in a DNA sample utilizing on-slide PCR gel-drop microarray technology.The TruArray® Warfarin Sensitivity Test Kit is an in vitro diagnostic test for the detection and genotyping of the 2C92, 2C93 alleles of the cytochrome P450 (CYP450) 2C9 gene locus and Vitamin K epoxide reductase CI, VKORCI, gene promoter polymorphism (-1639) variants from genomic DNA of human saliva samples collected using the Oragene® Dx Device (OGD-500) as an aid in the identification of patients at risk for increased warfarin sensitivity. The TruArray® Warfarin Sensitivity Test Kit is a qualitative assay for use in	eSensor Warfarin Sensitivity Test and XT-8 System (K073720)Same	The TruDiagnosis System and the eSensor Warfarin Sensitivity Test and XT-8 System have the same indications for use.
Item	Subject Device	Predicate Device	Comparison
	TruDiagnosis® System	eSensor Warfarin SensitivityTest and XT-8 System (K073720)
	clinical laboratories uponprescription by the attendingphysician.
Type of Use	Prescription Use	Prescription Use	The TruDiagnosis System is identicalto the eSensor Warfarin SensitivityTest and XT-8 System.
Technology	Fluorescence detection based gel-drop microarray	Electrochemical detection basedmicroarray	The TruDiagnosis System isequivalent to the eSensor WarfarinSensitivity Test and XT-8 System.Both systems employ the detectionof labeled probes on a microarray.The only difference is that theTruDiagnosis System uses afluorescent labeled probe forfluorescence detection, and theeSensor system uses a ferrocenelabeled probe for electrochemicaldetection. This difference in signaltransduction detection method doesnot does not affect the intended useor the safety and effectiveness of thedevice.
Detection Chemistry	Fluorogenic target-specificmicroarray-based hybridization.Signal detection using fluorescence.	The signal probe containselectrochemically active ferrocenelabels. Hybridization of the three-member complex at the electrodesurface and subsequentapplication of an excitation voltagecauses the ferrous ion in eachferrocene group to undergo cyclicoxidation and reduction at itscharacteristic redox potential,leading to loss or gain of an	The TruDiagnosis System is differentfrom the eSensor WarfarinSensitivity Test and XT-8 System.This difference does not affect theintended use or the safety andeffectiveness of the device.
Item	Subject Device	Predicate Device	Comparison
	TruDiagnosis® System	eSensor Warfarin SensitivityTest and XT-8 System (K073720)
		electron, and the generation of analternating current at the electrodesurface that is measured usingvoltammetry.
Signal Transduction	Fluorescence	Electrical current, voltammetry	The signal transduction of theTruDiagnosis System is differentfrom the signal transduction of theeSensor Warfarin Sensitivity Testand XT-8 System. This differencedoes not affect the intended use orthe safety and effectiveness of thedevice.
Test Type	Qualitative genetic test for singlenucleotide polymorphism detection.	Qualitative genetic test for singlenucleotide polymorphismdetection.	The TruDiagnosis System is identicalto the eSensor Warfarin SensitivityTest and XT-8 System.
Genes and SNPs	• CYP4502C92 (430C>T)• CYP4502C93 (1075A>C)• VKORC1 (-1639G>A)	• CYP2C92 (430C>T)• CYP2C93 (1075A>C)• VKORC1 (-1639G>A)	The TruDiagnosis System is identicalto the eSensor Warfarin SensitivityTest and XT-8 System.
Microarray-based genotypingtest for simultaneousdetection (multiplex system)of DNA sequences	Yes	Yes	The TruDiagnosis System isequivalent to the eSensor WarfarinSensitivity Test and XT-8 System.
Microarray Device	TruArray Test Slide	eSensor Test Cartridge	The TruDiagnosis System'smicroarray device is equivalent tothe eSensor Warfarin Sensitivity Testand XT-8 System's microarraydevice.
Assay result output	Assay results interpreted by asoftware program and assigned agenotype presented to the end userin a report format.	Assay results are interpreted by asoftware program and areassigned a result that is presentedto the end user in a report format.	The TruDiagnosis System isequivalent to the eSensor WarfarinSensitivity Test and XT-8 System.
Item	Subject Device	Predicate Device	Comparison
	TruDiagnosis® System	eSensor Warfarin SensitivityTest and XT-8 System (K073720)
Reference method	Bi-directional Sanger sequencing	Bi-directional Sanger sequencing	The TruDiagnosis System is identicalto the eSensor Warfarin SensitivityTest and XT-8 System.
Device Components	TruArray Test Slides, TruPlexReagents, and the TruDx 2000Imager	eSensor Test Cartridge,amplification and detectionreagents, and the eSensor XT-8System.	The components of the TruDiagnosisSystem are equivalent to thecomponents of the eSensor WarfarinSensitivity Test and XT-8 System.
Instrument	TruDx 2000 Imager	eSensor XT-8 System	The TruDiagnosis System includesan instrument equivalent to theinstrument included in the eSensorWarfarin Sensitivity Test and XT-8System.
Number of SimultaneousTests	1 test	8 tests	The TruDx 2000 Imager can onlyprocess one test at a time, while theXT-8 System can process multipletests simultaneously. This differencedoes not affect the intended use orthe safety and effectiveness of thedevice.
Test Processing Time	1 test within 30 seconds	8 tests within 40 minutes	The TruDx 2000 Imager is differentfrom the XT-8 System. Thisdifference in test processing timedoes not affect the intended use orsafety and effectiveness of thedevice.
User Interface	7" LCD touchscreen	15" LCD touchscreen	The user interface of the TruDxImager is equivalent to that of theeSensor XT-8 System. Bothinstruments include LCDtouchscreen interfaces. The onlydifference is that the screen size ofthe TruDx 2000 Imager is smallerthan that of the XT-8 System.However, this difference does not
Item	Subject Device	Predicate Device	Comparison
	TruDiagnosis® System	eSensor Warfarin SensitivityTest and XT-8 System (K073720)
			have an impact on the intended useor the safety and effectiveness of thedevice.
Power Requirements	Universal input, 90-264 Vac	100-240 Vac, 50/60 Hz, 600 W	The power requirements of TruDx2000 Imager and the eSensor XT-8System are equivalent. Both systemsare designed to use AC line power.
Dimensions	Approximately 10.93" x 8.5" x 16"	18.11" x 15.75" x 16.14"	The TruDx 2000 Imager is smallerthan the eSensor XT-8 System..This difference in size does not havean impact on the intended use or thesafety and effectiveness of thedevice.
Weight	Approximately 25 lbs	48.5 lbs	The TruDx 2000 Imager is lighterthan the eSensor XT-8 System.This difference in weight does nothave an impact on the intended useor the safety and effectiveness of thedevice.
Specimen type	Genomic DNA obtained fromepithelial cells contained in humansaliva samples collected using theOragene Dx Device OGD-500.	Genomic DNA obtained from ahuman whole blood sample.	The specimen type of theTruDiagnosis System is differentfrom the specimen type of theeSensor Warfarin Sensitivity Testand XT-8 System.This difference does not affect thesafety and effectiveness or intendeduse of the device.
Number of SNPs	3	3	The TruDiagnosis System is identicalto the eSensor Warfarin SensitivityTest and XT-8 System.
Item	Subject Device	Predicate Device	Comparison
	TruDiagnosis® System	eSensor Warfarin SensitivityTest and XT-8 System (K073720)
Input gDNA	25 ng of genomic DNA (5 µL of 5ng/µL genomic DNA sample)	10 ng of genomic DNA (5 µL of 2ng/µL genomic DNA sample)	The TruDiagnosis System is differentfrom the eSensor WarfarinSensitivity Test and XT-8 System.
DNA Extraction	Qiagen method. QIAAMP DNA MiniKit.	Manual ethanol extraction method	The TruDiagnosis System is differentfrom the eSensor WarfarinSensitivity Test and XT-8 SystemTest.This difference does not affect thesafety and effectiveness or intendeduse of the device.
PCR Process	Closed amplicon. On-slide PCRoccurs in the same chamber as themicroarray. Asymmetric PCR isperformed on-slide to eliminate thepredicate's exonuclease step.	Open amplicon. Two steps occuroff cartridge: 1) tube-based PCRand 2) exonuclease enzymetreatment to generate singlestranded target DNA.	The PCR Process of theTruDiagnosis System is differentfrom that of the eSensor WarfarinSensitivity Test and XT-8 System.This difference does not affect thesafety and effectiveness or intendeduse of the device.
Thermal Cycling Used	Yes	Yes	The TruDiagnosis System is identicalto the eSensor Warfarin SensitivityTest and XT-8 System.
Target of Detection	Single-nucleotide polymorphism	Single-nucleotide polymorphism	The TruDiagnosis System is identicalto the eSensor Warfarin SensitivityTest and XT-8 System.
Protocol Identification	Uses a barcode to store protocol andproduction information	Uses an internal memory chip(EEPROM) to store protocol andproduction information.	The TruDiagnosis System isequivalent to the eSensor WarfarinSensitivity Test and XT-8 System.

Detailed Comparison of the Subject and Predicate Devices:

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Substantial Equivalence Conclusions:

In conclusion, the TruDiagnosis System and the eSensor Warfarin Sensitivity Test and XT-8 System have the same indications for use related to aiding in the identification of patients at risk for increased warfarin sensitivity via genotyping multiple genetic variants in a DNA sample. The TruDx 2000 Imager and the eSensor XT-8 System have the same indications for use, in relation to the use of an instrument. The TruArray Warfarin Sensitivity Test Kit and the eSensor Warfarin Sensitivity Test have the same indications for use, related to Warfarin sensitivity genotyping of the *2 (430C>T) and * 3 (1075A>C) 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. The technological characteristics and testing demonstrate that the TruDiagnosis System is substantially equivalent to the eSensor Waffarin Sensitivity Test and XT-8 System and assures that the TruDiagnosis System is as safe and effective as the eSensor Warfarin Sensitivity Test and XT-8 System.

Performance Characteristics

A series of studies have been performed to validate the performance characteristics for the intended use of the TruDiagnosis® System. A total of five (5) performance characteristics studies have been conducted and two Stability Testing Studies were conducted to provide data on the kit configuration and expiry of the kit. A summary of each study's results is provided below.

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Limit of Detection

An upper and lower limit of detection study was performed to assess the TruArray® Warfarin Sensitivity Test Kit across a range of genomic DNA input concentrations. In order to determine the lowest concentration of DNA at which this assay is acurate, serial diutions (125, 25, 0.25 ng) of thee genomic DNA samples were assayed 40 times using the TruArray® Warfarin Sensitivity Test Kit. Two entire runs were repeated as a result of control failures and two additional runs were performed for individual test samples that gave a first-pass no-call result. The study was performed with reagents and tests from 5 lots of the TruArray® Warfarin Sensitivity Test Kit, using three TruDx® 2000 Imager instruments, and 7 operators over the course of 20 total calendar days. A total of 362 samples were tested and all genotyping calls of the Trukrray® Warfarin Sensitivity Test Kit were compared to their corollary bi-directional Sanger sequencing result. The recommended DNA input for the TruArray® Waffain Sensitivity Test Kit is 25 ng at a concentration of 5 ng/uL.

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Limit of Detection Study by Sample Genotype

		Genotype				First Pass Calls						Final Resulta
Donor	2C9*2	2C9*3	VKORC1	DNAInput(ng)	#ReplicatesTested	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRateb	95%LCB	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRateb	95%LCB
1	1/1	1/3	AA	125	40	40	0	0	100.0%	92.8%	40	0	0	100.0%	92.8%
1	1/1	1/3	AA	25	40	39	1	0	97.5%	88.7%	40	0	0	100.0%	92.8%
1	1/1	1/3	AA	2.5	40	39	1	0	97.5%	88.7%	40	0	0	100.0%	92.8%
1	1/1	1/3	AA	0.25c	40	28	12	0	70.0%	56.0%	28	12	0	70.0%	56.0%
1	1/1	1/3	AA	2.5-125	120	118	2	0	98.3%	94.8%	120	0	0	100.0%	97.5%
2	1/2	1/3	AG	125	40	28	12d	0	70.0%	56.0%	40	0	0	100.0%	92.8%
2	1/2	1/3	AG	25	40	40	0	0	100.0%	92.8%	40	0	0	100.0%	92.8%
2	1/2	1/3	AG	2.5	40	35	5	0	87.5%	75.5%	40	0	0	100.0%	92.8%
2	1/2	1/3	AG	0.25c	40	15	22	3	37.5%	24.7%	15	22	3	37.5%	24.7%
2	1/2	1/3	AG	2.5-125	120	103	17	0	85.8%	79.5%	120	0	0	100.0%	97.5%
3	2/2	1/1	GG	125	42	25	17d	0	59.5%	45.7%	40	0	0	100.0%	92.8%
3	2/2	1/1	GG	25	40	37	3	0	92.5%	81.7%	40	0	0	100.0%	92.8%
3	2/2	1/1	GG	2.5	40	36	4	0	90.0%	78.6%	40	0	0	100.0%	92.8%
3	2/2	1/1	GG	0.25c	41	3	38	0	7.3%	2.0%	3	38	0	7.3%	2.0%
3	2/2	1/1	GG	2.5-125	122	98	24	0	80.3%	73.5%	120	0	0	100.0%	97.5%
			Total all 3 Samples	2.5-125	362	319	43d	0	88.1%	85.0%	360	0	0	100.0%	99.2%

ª Final Result represents a second repeat of a No Call. Incorrect Calls were not repeated.

ካ The Correct Call rate is the proportion of Correct Calls from the total number of calls.

° Repeat runs were not performed.

d 2 runs had a negative control failure.

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Limit of Detection Study by Loci

					First Pass Calls		Final Result a
DNAAmount(ng)	Loci	TotalNumberof Calls	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRate b	95%LCB	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRate b	95%LCB
125	2C9*2	122	93	29 c	0	76.2%	69.0%	122	0	0	100.0%	97.6%
125	2C9*3	122	96	26 c	0	78.7%	71.7%	122	0	0	100.0%	97.6%
125	VKORC1	122	96	26 c	0	78.7%	71.7%	122	0	0	100.0%	97.6%
125	TOTAL	366	360	6 c	0	98.4%	96.8%	366	0	0	100.0%	99.2%
25	2C9*2	120	119	1	0	99.2%	96.1%	120	0	0	100.0%	97.5%
25	2C9*3	120	119	1	0	99.2%	96.1%	120	0	0	100.0%	97.5%
25	VKORC1	120	116	4	0	96.7%	92.5%	120	0	0	100.0%	97.5%
25	TOTAL	360	354	6	0	98.3%	96.7%	360	0	0	100.0%	99.2%
2.5	2C9*2	120	115	5	0	95.8%	91.4%	120	0	0	100.0%	97.5%
2.5	2C9*3	120	112	8	0	93.3%	88.3%	120	0	0	100.0%	97.5%
2.5	VKORC1	120	113	7	0	94.2%	89.3%	120	0	0	100.0%	97.5%
2.5	TOTAL	360	340	20	0	94.4%	92.0%	360	0	0	100.0%	99.2%
0.25 c	2C9*2	121	86	34	1	71.1%	63.5%	85	34	1	70.2%	62.7%
0.25 c	2C9*3	121	73	46	2	60.3%	52.5%	72	46	2	59.5%	51.6%
0.25 c	VKORC1	121	73	48	0	60.3%	52.5%	72	48	0	59.5%	51.6%
0.25 c	TOTAL	363	235	128	0	64.7%	60.4%	232	128	0	63.9%	59.6%

ª Final Result represents a second repeat of a No Call. Incorrect Calls were not repeated.

ఠ The Correct Call rate is the proportion of Correct Calls from the total number of calls.

' Repeat runs were not performed.

d 2 runs had a negative control failure.

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Method Comparison

The method comparison evaluation was performed to assess the genotyping performance of the TruArray® Warfarin Sensitivity Test Kit as compared to bi-directional DNA sequencing across an extensive range of donor genotypes. Testing was conducted at one site (internal site). The test panel for the method consisted of 303 unique human genomic DNA samples isolated from saliva specimens collected using the Oragene® Dx OGD-500 device followed by DNA extraction using the Qiagen QAA Mini Kit and genotype testing using the TruArray® Warfarin Sensitivity Test Kit.

The study was performed with four lots of the Truly Warfarin Sensitivity Test Kit, four TruDx® 2000 Imager instruments, and four operators over 14 testing days over the course of 30 total consecutive calendar days. The samples were typically run for their firstpass call in batches of 16 samples, a neqative extraction control, a positive PCR control and negative PCR control. An additional run was performed for tests that qave a first-pass no-call result.

Results are as summarized in the tables below. All genotyping calls of the TruArray® Warfarin Sensitivity Test Kit were compared to their corollary bi-directional sequencing result. For the first pass runs, a total of 303 donor samples were run resulting in 299 Correct Calls, 3 No Calls, and 1 Incorrect Calls. An investigation revealed that the incorrect call was the result of a sample mix-up. All 3 No Calls were resolved upon the second repeat runs. The first-pass runs is 98.7% and a 95% lower confidence bound of 97.0%. For the repeat runs (Final Result), the Correct Call Rate is 100.0% and a 95.0%. The Final Results yielded 100% agreement between the TruArray® Warfarin Sensitivity Test Kit results and bi-directional DNA sequencing.

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	Bi-directional SequencingSample Genotype				First Pass Calls		Final Result a
2C9*2	2C9*3	VKORC1	#SamplesTested	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRate b	95%LCB	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRate b	95%LCB
1/1	1/1	AA	58	57	1	0	98.3%	92.1%	58	0	0	100.0%	95.0%
1/1	1/1	GA	79	79	0	0	100.0%	96.3%	79	0	0	100.0%	96.3%
1/1	1/1	GG	41	41	0	0	100.0%	93.0%	41	0	0	100.0%	93.0%
1/2	1/1	AA	8	8	0	0	100.0%	68.8%	8	0	0	100.0%	68.8%
1/2	1/1	GA	21	21	0	0	100.0%	86.7%	21	0	0	100.0%	86.7%
1/2	1/1	GG	30	29	1	0	96.7%	85.1%	30	0	0	100.0%	90.5%
1/1	1/3	AA	8	7	1	0	87.5%	52.9%	8	0	0	100.0%	68.8%
1/1	1/3	GA	15	15	0	0	100.0%	81.9%	15	0	0	100.0%	81.9%
1/1	1/3	GG	25	25	0	0	100.0%	88.7%	25	0	0	100.0%	88.7%
2/2	1/1	AA	1	1	0	0	100.0%	5.0%	1	0	0	100.0%	5.0%
2/2	1/1	GA	6	5	0	1 c	83.3%	41.8%	6	0	1 c	83.3%	41.8%
2/2	1/1	GG	3	3	0	0	100.0%	36.8%	3	0	0	100.0%	36.8%
1/1	3/3	GG	1	1	0	0	100.0%	5.0%	1	0	0	100.0%	5.0%
1/2	1/3	GA	6	6	0	0	100.0%	60.7%	6	0	0	100.0%	60.7%
1/2	1/3	GG	1	1	0	0	100.0%	5.0%	1	0	0	100.0%	5.0%
	Total		303	299	3	1 c	98.7%	97.0%	302	0	1 c	99.7%	98.4%

Agreement between TruArray® Warfarin Sensitivity Test Kit and Bi-directional DNA Sequencing (by Genotype):

ª Final Result represents a second repeat of a No Call. Incorrect Calls were not repeated.

ఠ The Correct Call rate is the proportion of Correct Calls from the total number of calls.

^ One sample reported an incorrect call in the first run. Upon investigation, it was determined a sample mix in efirst-pass call matched the bi-directional DNA sequencing result for the correctly accessioned samples. This sample was not re-run.

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Bi-directional SequencingSample Genotype			First Pass Calls						Final Result a
Loci	Genotype	#SamplesTested	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRate b	95%LCB	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRate b	95%LCB
2C9*2	wt/wt	227	227	0	0	100.0%	98.7%	227	0	0	100.0%	98.7%
2C9*2	wt/*2	66	66	0	0	100.0%	95.6%	66	0	0	100.0%	95.6%
2C9*2	2/2	10	10	0	0	100.0%	74.1%	10	0	0	100.0%	74.1%
2C9*3	wt/wt	247	247	0	0	100.0%	98.8%	247	0	0	100.0%	98.8%
2C9*3	wt/*3	55	54	1	0	98.2%	91.7%	55	0	0	100.0%	94.7%
2C9*3	3/3	1	1	0	0	100.0%	5.0%	1	0	0	100.0%	5.0%
VKORC1 (-1639)	GG	102	101	1	0	99.0%	95.4%	102	0	0	100.0%	97.1%
VKORC1 (-1639)	GA	126	125	0	1 c	99.2%	96.3%	125	0	1 c	99.2%	96.3%
VKORC1 (-1639)	AA	75	74	1	0	98.7%	93.8%	75	0	0	100.0%	96.1%

Agreement between TruArray® Warfarin Sensitivity Test Kit and Bi-directional DNA Sequencing (by Loci):

ª Final Result represents a second repeat of a No Call. Incorrect Calls were not repeated.

ካ The Correct Call rate is the proportion of Correct Calls from the total number of calls.

^ One sample reported an incorrect call in the first run. Upon investigation, it was determined a sample mix a me first-pass call matched the bi-directional DNA sequencing result for the correctly accessioned samples. This sample was not re-run.

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Interference Substances

Effect of Endogenous Interfering Substances:

Interfering substances including salivase, hemoglobin A (lgA) and total protein were spiked into saliva samples in the amounts used by the predicate (GenMark, K110786). Saliva samples were collected using the OrageneDx OGD-500 device and extracted using the Qiagen QlA Mini Kt. Five donors provided saliva samples, each of which were divided into five aliquots and spiked with one of the fith aliquot remained un-spiked and served as a control. Three extractions were performed on each spiked and un-spiked sample.

The study was performed with reagents and tests from 3 lots of the TruArray® Warfarin Sensitivity Test Kit, using two TruDx® 2000 Imager instruments, and two operators over 5 testing days over the course of 8 total calendar days.

All genotyping calls of the TruArray® Warfarin Sensitivity Test Kit were compared to their corollary bi-directional sequencing result. The first run for the study resulted in 3 No Calls which were resolved upon the repeat run. The Final Results yielded 100% agreement between the TruArray® Warfarin Sensitivity Test Kit results and bi-directional DNA sequencing for all test substances demonstrating no effect of any interfering substances on genotyping.

InterferenceCondition	InterferenceConcentration	#SamplesTested	#CorrectCalls	#NoCalls	CorrectCallRate b	95%LCB	#CorrectCalls	CorrectCallRate b	95%LCB
Control Total	N/A	15	15	0	100.0%	81.9%	15	100.0%	81.9%
Amylase Total	311 - 341 U	15	15	0	100.0%	81.9%	15	100.0%	81.9%
HemoglobinTotal	21.2 - 22.1 mg/mL	15	14	1	93.3%	72.1%	15	100.0%	81.9%
IgA Total	0.3 - 0.5 mg/mL	15	15	0	100.0%	81.9%	15	100.0%	81.9%
Protein Total	2.1 - 2.7 mg/mL	15	13	2	86.7%	63.7%	15	100.0%	81.9%

Endogenous Interference

ª Final Result represents a second repeat of a No Call.

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b The Correct Call rate is the proportion of Correct Calls from the total number of calls.

Effect of Exogenous Interfering Substances:

Potentially interfering exogenous substances into saliva samples through various activities (eating, drinking, chewing gum, mouthwash and smoking) were tested. Each activity group was composed of 5 donors who each provided 2 saliva samples - a baseline/control sample prior to the activity, and a sample collected 30 minutes after the activity. Samples were tested in triplicate. Saliva samples were collected using the OrageneDx OGD-500 device and extracted using the Qiagen QAamp DSP DNA Mini Kit. A total of 150 samples were tested and all genotyping calls of the TruArray® Warfarin Sensitivity Test Kit were compared to their corollary bi-directional Sanger sequencing result.

Activity	Time-point	# Samples Tested	# Correct Calls	# No Calls	Correct Call Rate b	95% LCB	# Correct Calls	Correct Call Rate b	95% LCB
Eating	Baseline	15	14	1	93.3%	72.1%	15	100.0%	81.9%
Eating	30 min.	15	15	0	100.0%	81.9%	15	100.0%	81.9%
Drinking	Baseline	15	15	0	100.0%	81.9%	15	100.0%	81.9%
Drinking	30 min.	15	15	0	100.0%	81.9%	15	100.0%	81.9%
Chewing Gum	Baseline	15	15	0	100.0%	81.9%	15	100.0%	81.9%
Chewing Gum	30 min.	15	13	2	86.7%	63.7%	15	100.0%	81.9%
Mouth Wash	Baseline	15	15	0	100.0%	81.9%	15	100.0%	81.9%
Mouth Wash	30 min.	15	15	0	100.0%	81.9%	15	100.0%	81.9%
Smoking	Baseline	15	6	9	40.0%	19.0%	15	100.0%	81.9%
Smoking	30 min.	15	9	6	60.0%	36.0%	15	100.0%	81.9%
Total	Baseline	75	65	10	86.7%	78.4%	75	100.0%	96.1%
Total	30 min.	75	67	8	89.3%	81.6%	75	100.0%	96.1%

Exogenous Interference

ª Final Result represents a second repeat of a No Call.

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b The Correct Call rate is the proportion of Correct Calls from the total number of calls.

0 1 run in this set had a negative control failure. The smoking calls and 5 post-smoking calls which were all Correct Calls when repeated.

Reproducibility

A reproducibility study was performed to assess the variability of the TruArray® Warfarin Sensitivity Test Kit between clinical study sites, operators, days, sample genotypes, instruments, and different lots of TruArray® Warfarin Sensitivity Test Kit. Six specific saliva samples, covering all genotypes detected by the TruArray® Warfarin Sensitivity Test Kit were used for this study. Saliva samples were collected using the OrageneDx OGD-500 device and extracted using the Qiagen QlAamp DSP DNA Mini Kit. The study was performed at 3 sites with 4 different lots of the TruArray® Warfarin Sensitivity Test Kit, using 4 TruDx™ 2000 Imager instruments, 4 ProFlex™ PCR Systems using the ProFlex™ 2x Flat Sample Block, 6 different genotype saliva samples, and 4 operators testing over 20 nonconsecutive. Site 1 (internal) had two operators and Site 3 each had one operator for a study total of four operators. Each operator processed a single (genotype) in triplicate and repeated over 5 non-consecutive days (15 tests per operator per qenotype). Four operators generated 60 data points per genotype (4 operators x 15 tests per operator per genotype). 6 different sample genotypes generated 90 data points per operator (6 genotypes x 15 tests per operator per genotype). A total of 360 samples were tested and all genotyping calls of the TruArray® Warfarin Sensitivity Test Kit were compared to their corollary bidirectional Sanger sequencing result.

Sequencing Genotype			# Samples Tested	First Pass Results			Final Resultsa
2C9*2	2C9*3	VKORC1		# Correct Calls	# No Calls	# Incorrect Calls	# Correct Calls	# No Calls	# Incorrect Calls	Correct Call Rateb	95% LCB
1/1	3/3	GG	60	52	8	0	86.7%	77.2%	60	0	0	100.0%	95.1%
1/2	1/1	AA	60	53	7	0	88.3%	79.2%	60	0	0	100.0%	95.1%
1/1	1/1	GG	60	50	10	0	83.3%	73.4%	60	0	0	100.0%	95.1%
1/1	1/3	AA	60	50	10	0	83.3%	73.4%	60	0	0	100.0%	95.1%
2/2	1/1	GA	60	50	10	0	83.3%	73.4%	60	0	0	100.0%	95.1%
1/2	1/3	GA	60	48	12	0	80.0%	69.6%	60	0	0	100.0%	95.1%
Total			360	303	57	0	84.2%	80.7%	360	0	0	100.0%	99.2%

Reproducibility Study by Sample Genotype

ª Final Result represents up to four repeats of a No Call.

{22}------------------------------------------------

ካ The Correct Call rate is the proportion of Correct Calls from the total number of calls.

{23}------------------------------------------------

Reproducibility Study by Site, Operator and Loci

Site	Operator	Loci	#SamplesTested	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRate b	95%LCB	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRate b	95%LCB
First Pass Results								Final Results a
1	1	2C9*2	90	78	12	0	86.7%	79.3%	90	0	0	100.0%	96.7%
1	1	2C9*3	90	79	11	0	87.8%	80.6%	90	0	0	100.0%	96.7%
1	1	VKORC1	90	79	11	0	87.8%	80.6%	90	0	0	100.0%	96.7%
1	2	2C9*2	90	82	8	0	91.1%	84.5%	90	0	0	100.0%	96.7%
1	2	2C9*3	90	79	11	0	87.8%	80.6%	90	0	0	100.0%	96.7%
1	2	VKORC1	90	80	10	0	88.9%	81.9%	90	0	0	100.0%	96.7%
2	3	2C9*2	90	71	19	0	78.9%	70.6%	90	0	0	100.0%	96.7%
2	3	2C9*3	90	70	20	0	77.8%	69.4%	90	0	0	100.0%	96.7%
2	3	VKORC1	90	71	19	0	78.9%	70.6%	90	0	0	100.0%	96.7%
3	4	2C9*2	90	80	10	0	88.9%	81.9%	90	0	0	100.0%	96.7%
3	4	2C9*3	90	80	10	0	88.9%	81.9%	90	0	0	100.0%	96.7%
3	4	VKORC1	90	78	12	0	86.7%	79.3%	90	0	0	100.0%	96.7%
Total	Loci	2C9*2	360	311	49	0	86.4%	83.1%	360	0	0	100.0%	99.2%
Total	Loci	2C9*3	360	308	52	0	85.6%	82.2%	360	0	0	100.0%	99.2%
Total	Loci	VKORC1	360	308	52	0	85.6%	82.2%	360	0	0	100.0%	99.2%
Total	SNPs		1080	927	153	0	85.8%	84.0%	1080	0	0	100.0%	99.7%

ª Final Result represents up to four repeats of a No Call.

ካ The Correct Call rate is the proportion of Correct Calls from the total number of calls.

{24}------------------------------------------------

Reproducibility Study by Production Lot and Loci

		First Pass Results					Final Results "
Lot	Loci	#SamplesTested	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRate b	95%LCB	#CorrectCalls	#NoCalls	#IncorrectCalls	CorrectCallRate b	95%LCB
#15007/5008	2C9*2	72	71	1	0	98.6%	93.6%	72	0	0	100.0%	95.9%
	2C9*3	72	71	1	0	98.6%	93.6%	72	0	0	100.0%	95.9%
	VKORC1	72	70	2	0	97.2%	91.5%	72	0	0	100.0%	95.9%
#20011/0008	2C9*2	108	87	21	0	80.6%	73.2%	108	0	0	100.0%	97.3%
	2C9*3	108	87	21	0	80.6%	73.2%	108	0	0	100.0%	97.3%
	VKORC1	108	87	21	0	80.6%	73.2%	108	0	0	100.0%	97.3%
#38005/8004	2C9*2	108	98	10	0	90.7%	84.8%	108	0	0	100.0%	97.3%
	2C9*3	108	96	12	0	88.9%	82.6%	108	0	0	100.0%	97.3%
	VKORC1	108	96	12	0	88.9%	82.6%	108	0	0	100.0%	97.3%
#46010/0015	2C9*2	72	55	17	0	76.4%	66.7%	72	0	0	100.0%	95.9%
	2C9*3	72	54	18	0	75.0%	65.2%	72	0	0	100.0%	95.9%
	VKORC1	72	55	17	0	76.4%	66.7%	72	0	0	100.0%	95.9%
AllLots	2C9*2	360	311	49	0	86.4%	83.1%	360	0	0	100.0%	99.2%
	2C9*3	360	308	52	0	85.6%	82.2%	360	0	0	100.0%	99.2%
	VKORC1	360	308	52	0	85.6%	82.2%	360	0	0	100.0%	99.2%
SNPs		1080	927	153	0	85.8%	84.0%	1080	0	0	100.0%	99.7%

ª Final Result represents up to four repeats of a No Call.

ካ The Correct Call rate is the proportion of Correct Calls from the total number of calls.

Kit Stability

Akonni conducted a Stability study to determine the expiration date of the TruArray® Warfarin Sensitivity Test Kit. Based on this 9 months of Stability testing, an expiry date of 6 months was determined for the TruArray® Warfarin Sensitivity Test Kit.

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EMC and Safety Testing

Safety testing was performed on the TruDiagnosis® System in accordance with the following Standard:

IEC 61010-1:2010 3rd edition: General Requirements for Basic Safety and Essential Performance ●
EN 61326-1:2013 / EN 55011:2010 / EN 55022:2010: Electromagnetic Compatibility (EMC) ●
IEC 60601-1-2:2014 Medical electrical equipment Part 1-2: General requirements for basic safety and essential . performance - Collateral standard: Electromagnetic compatibility - Requirements and tests

TruDx® Imager and TruSpot™ Software Performance Testing

Vertication testing for the TruDx® 2000 Imager was performed to confirm the documented Product Requirements. A total of eight (8) different verification tests were performed, as listed below.

Mechanical Verification ●
Electrical Verification ●
Functional Verification ●
Shipping and Packaging ●
Environmental Verification ●
Reliability Verification ●
Hardware Interface Verification ●
Performance Verification ●
Software Verification ●

TruDx® Imager and TruSpot™ Software were validated in accordance with a Validation plan to ensure conformance with established performance criteria.

Conclusion

The 510(k) Pre-market Notification for the TruDiagnosis® System contains adequate information and data to determine that Akonni's TruDiaqnosis® System is as safe and effective as the legally marketed predicate device(s).

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.