Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The document describes standard image processing and analysis techniques for microarray data, but does not mention AI or ML.

Therapeutic

No
This device is an in vitro diagnostic (IVD) device used for genotyping genetic variants to aid in the identification of patients at risk for increased warfarin sensitivity. It does not provide direct therapy to a patient.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states, "The TruDiagnosis® System is an in vitro diagnostic device" and describes its purpose in processing and genotyping DNA samples for identifying genetic variants related to warfarin sensitivity.

SaMD (Software as a Medical Device)

No

The device is described as a "System" that includes hardware components (TruDx® 2000 Imager, ProFlex™ PCR System), a test kit, and software (TruSpot™ Software). The software is integrated within the imager and analyzes images captured by the hardware. This is not a software-only medical device.

IVD (In Vitro Diagnostic)

Yes, the provided text explicitly states that the TruDiagnosis® System is an In Vitro Diagnostic (IVD) device.

Here are the key phrases from the text that confirm this:

"The TruDiagnosis® System is an in vitro diagnostic device intended for processing and genotyping multiple genetic variants..." (from the Intended Use/Indications for Use section)
"The TruDiagnosis® System is an in vitro diagnostic device intended for processing and genotyping multiple genes in a DNA sample..." (from the Device Description section)
"The TruArray® Warfarin Sensitivity Test Kit is an in vitro diagnostic test for the detection and genotyping of the 2C92, 2C93 alleles..." (from the Intended Use/Indications for Use section)
"The warfarin assay performed with the test kit is an in vitro diagnostic test for the detection and genotyping of the 2C92, 2C93 alleles..." (from the Device Description section)

These statements clearly identify the system and its key components as IVD devices.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

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.

Product codes (comma separated list FDA assigned to the subject device)

ODW, ODV, NSU

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
- 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.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Clinical laboratories upon prescription by the attending physician.

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Not Found

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Limit of Detection

Study Type: Accuracy determination at various DNA input concentrations.
Sample Size: 362 samples tested (3 human genomic DNA samples with serial dilutions at 125, 25, 2.5, and 0.25 ng, each assayed 40 times).
Key Results:
- The lowest concentration of DNA at which the assay is accurate.
- All genotyping calls of the TruArray® Warfarin Sensitivity Test Kit were compared to their corollary bi-directional Sanger sequencing result.
- The recommended DNA input for the TruArray® Warfarin Sensitivity Test Kit is 25 ng at a concentration of 5 ng/uL.
- At 25 ng input, the Correct Call Rate was 98.3% (96.7% 95% LCB) for first pass calls and 100.0% (99.2% 95% LCB) for final results across all 360 calls for all samples.
- For individual loci at 25 ng: 2C92 (99.2% first pass, 100.0% final), 2C93 (99.2% first pass, 100.0% final), VKORC1 (96.7% first pass, 100.0% final).
- At 0.25 ng input, the Correct Call Rate was 64.7% (60.4% 95% LCB) for first pass calls and 63.9% (59.6% 95% LCB) for final results across all 363 calls.

Method Comparison

Study Type: Genotyping performance comparison against bi-directional DNA sequencing.
Sample Size: 303 unique human genomic DNA samples.
Data Source: Samples isolated from saliva specimens collected using the Oragene® Dx OGD-500 device, followed by DNA extraction using the Qiagen QIAamp Mini Kit.
Key Results:
- All genotyping calls of the TruArray® Warfarin Sensitivity Test Kit were compared to their corollary bi-directional sequencing result.
- For first pass runs (303 donor samples): 299 Correct Calls, 3 No Calls, 1 Incorrect Call. The first-pass Correct Call Rate was 98.7% with a 95% lower confidence bound of 97.0%.
- For repeat runs (Final Result): The Correct Call Rate was 100.0% with a 95% lower confidence bound of 95.0%.
- The Final Results yielded 100% agreement between the TruArray® Warfarin Sensitivity Test Kit results and bi-directional DNA sequencing, with one incorrect call attributed to a sample mix-up that was not re-run.

Interference Substances (Endogenous)

Study Type: Effect of endogenous interfering substances (salivase, hemoglobin A (IgA), total protein) on genotyping.
Sample Size: 5 donors, each with 5 aliquots (4 spiked, 1 control), with 3 extractions performed on each sample; total of 75 samples (5 donors x 5 aliquots x 3 extractions).
Key Results:
- All genotyping calls of the TruArray® Warfarin Sensitivity Test Kit were compared to their corollary bi-directional sequencing result.
- The first run resulted in 3 No Calls, which were resolved upon repeat runs.
- 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.

Interference Substances (Exogenous)

Study Type: Effect of exogenous interfering substances (eating, drinking, chewing gum, mouthwash, smoking) on genotyping.
Sample Size: 5 donors per activity group, each providing 2 saliva samples (baseline/control and 30 mins after activity), tested in triplicate; total of 150 samples.
Key Results:
- All genotyping calls of the TruArray® Warfarin Sensitivity Test Kit were compared to their corollary bi-directional Sanger sequencing result.
- For first pass: Total Correct Call Rate was 86.7% (78.4% LCB) at baseline and 89.3% (81.6% LCB) at 30 min.
- For final results: All activities and time-points yielded 100.0% Correct Call Rate (96.1% LCB for total).

Reproducibility

Study Type: Assessment of variability across sites, operators, days, sample genotypes, instruments, and kit lots.
Sample Size: 6 specific saliva samples covering all detectable genotypes. Total of 360 samples tested (6 genotypes x 4 operators x 15 tests/operator/genotype).
Data Source: Saliva samples collected using the OrageneDx OGD-500 device and extracted using the Qiagen QIAamp DSP DNA Mini Kit.
Key Results:
- All genotyping calls were compared to bi-directional Sanger sequencing result.
- By Sample Genotype: First pass Correct Call Rate ranged from 80.0% to 88.3%. Final Result Correct Call Rate was 100.0% across all 6 genotypes.
- By Site, Operator, and Loci: First pass Correct Call Rate for loci varied from 77.8% to 91.1%. Final Result Correct Call Rate was 100.0% for all sites, operators, and loci.
- By Production Lot and Loci: First pass Correct Call Rate for loci varied from 75.0% to 98.6%. Final Result Correct Call Rate was 100.0% for all production lots and loci.

Kit Stability

Study Type: Determination of the expiration date for the TruArray® Warfarin Sensitivity Test Kit.
Key Results: Based on 9 months of stability testing, an expiry date of 6 months was determined for the kit.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Correct Call Rate:
- Limit of Detection Study:
  - At 25 ng input (Final Result): 100.0% (99.2% 95% LCB)
  - At 0.25 ng input (Final Result): 63.9% (59.6% 95% LCB)
- Method Comparison Study:
  - First Pass: 98.7% (97.0% 95% LCB)
  - Final Result: 100.0% (95.0% 95% LCB)
- Interference Substances (Endogenous): Final Result: 100.0% (81.9% 95% LCB) for all substances.
- Interference Substances (Exogenous): Final Result: 100.0% (96.1% 95% LCB for total).
- Reproducibility Study: Final Result: 100.0% (99.2% 95% LCB for total samples; 99.7% 95% LCB for total SNPs)

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

K073720

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

K110701

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

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.

Summary

0

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

1

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

2

Indications for Use

510(k) Number (if known) K183530

Device Name TruDiagnosis® System

Indications for Use (Describe)

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.

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)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

3

510(k) Summary

Akonni Biosystems, Inc. (K183530)

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

| Submitter: | Akonni Biosystems, Inc.
400 Sagner Avenue
Suite 300
Frederick, MD 21701
Phone: (301) 698-0101
Fax: (301) 698-0202 |
|--------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Primary Contact: | Rita King, CEO
MethodSense, Inc.
Email: ritaking@methodsense.com
Phone: (919) 313-3961
Fax: (919) 313-3979 |
| Company Contact: | Charles Daitch
CEO |
| 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 Genotyping
Systems
21 CFR §864.7750 Prothrombin time test
21 CFR §862.2570 Instrumentation for Clinical Multiplex
Test Systems |
| Classification Panel: | Toxicology (91), Hematology (81), Chemistry (75) |
| Product Code: | ODW- Cytochrom P450 2C9 (CYP450 2C9) Drug Metabolizing
Enzyme Genotyping System
ODV- Vitamin K epoxide reductase complex subunit 1 (VKORC1)
Genotyping System
NSU- 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 Genotyping
Test |

4

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

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.

5

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.

Indications for Use

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, 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.

6

Item	Subject Device	Predicate Device	Comparison
Indications for Use	TruDiagnosis® System
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 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 Sensitivity
Test and XT-8 System (K073720) | |
| | clinical laboratories upon
prescription by the attending
physician. | | |
| Type of Use | Prescription Use | Prescription Use | The TruDiagnosis System is identical
to the eSensor Warfarin Sensitivity
Test and XT-8 System. |
| Technology | Fluorescence detection based gel-
drop microarray | Electrochemical detection based
microarray | The TruDiagnosis System is
equivalent to the eSensor Warfarin
Sensitivity Test and XT-8 System.
Both systems employ the detection
of labeled probes on a microarray.

The only difference is that the
TruDiagnosis System uses a
fluorescent labeled probe for
fluorescence detection, and the
eSensor system uses a ferrocene
labeled probe for electrochemical
detection. This difference in signal
transduction detection method does
not does not affect the intended use
or the safety and effectiveness of the
device. |
| Detection Chemistry | Fluorogenic target-specific
microarray-based hybridization.
Signal detection using fluorescence. | The signal probe contains
electrochemically active ferrocene
labels. Hybridization of the three-
member complex at the electrode
surface and subsequent
application of an excitation voltage
causes the ferrous ion in each
ferrocene group to undergo cyclic
oxidation and reduction at its
characteristic redox potential,
leading to loss or gain of an | The TruDiagnosis System is different
from the eSensor Warfarin
Sensitivity Test and XT-8 System.

This difference does not affect the
intended use or the safety and
effectiveness of the device. |
| Item | Subject Device | Predicate Device | Comparison |
| | TruDiagnosis® System | eSensor Warfarin Sensitivity
Test and XT-8 System (K073720) | |
| | | electron, and the generation of an
alternating current at the electrode
surface that is measured using
voltammetry. | |
| Signal Transduction | Fluorescence | Electrical current, voltammetry | The signal transduction of the
TruDiagnosis System is different
from the signal transduction of the
eSensor Warfarin Sensitivity Test
and XT-8 System. This difference
does not affect the intended use or
the safety and effectiveness of the
device. |
| Test Type | Qualitative genetic test for single
nucleotide polymorphism detection. | Qualitative genetic test for single
nucleotide polymorphism
detection. | The TruDiagnosis System is identical
to the eSensor Warfarin Sensitivity
Test 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 identical
to the eSensor Warfarin Sensitivity
Test and XT-8 System. |
| Microarray-based genotyping
test for simultaneous
detection (multiplex system)
of DNA sequences | Yes | Yes | The TruDiagnosis System is
equivalent to the eSensor Warfarin
Sensitivity Test and XT-8 System. |
| Microarray Device | TruArray Test Slide | eSensor Test Cartridge | The TruDiagnosis System's
microarray device is equivalent to
the eSensor Warfarin Sensitivity Test
and XT-8 System's microarray
device. |
| Assay result output | Assay results interpreted by a
software program and assigned a
genotype presented to the end user
in a report format. | Assay results are interpreted by a
software program and are
assigned a result that is presented
to the end user in a report format. | The TruDiagnosis System is
equivalent to the eSensor Warfarin
Sensitivity Test and XT-8 System. |
| Item | Subject Device | Predicate Device | Comparison |
| | TruDiagnosis® System | eSensor Warfarin Sensitivity
Test and XT-8 System (K073720) | |
| Reference method | Bi-directional Sanger sequencing | Bi-directional Sanger sequencing | The TruDiagnosis System is identical
to the eSensor Warfarin Sensitivity
Test and XT-8 System. |
| Device Components | TruArray Test Slides, TruPlex
Reagents, and the TruDx 2000
Imager | eSensor Test Cartridge,
amplification and detection
reagents, and the eSensor XT-8
System. | The components of the TruDiagnosis
System are equivalent to the
components of the eSensor Warfarin
Sensitivity Test and XT-8 System. |
| Instrument | TruDx 2000 Imager | eSensor XT-8 System | The TruDiagnosis System includes
an instrument equivalent to the
instrument included in the eSensor
Warfarin Sensitivity Test and XT-8
System. |
| Number of Simultaneous
Tests | 1 test | 8 tests | The TruDx 2000 Imager can only
process one test at a time, while the
XT-8 System can process multiple
tests simultaneously. This difference
does not affect the intended use or
the safety and effectiveness of the
device. |
| Test Processing Time | 1 test within 30 seconds | 8 tests within 40 minutes | The TruDx 2000 Imager is different
from the XT-8 System. This
difference in test processing time
does not affect the intended use or
safety and effectiveness of the
device. |
| User Interface | 7" LCD touchscreen | 15" LCD touchscreen | The user interface of the TruDx
Imager is equivalent to that of the
eSensor XT-8 System. Both
instruments include LCD
touchscreen interfaces. The only
difference is that the screen size of
the TruDx 2000 Imager is smaller
than that of the XT-8 System.
However, this difference does not |
| Item | Subject Device | Predicate Device | Comparison |
| | TruDiagnosis® System | eSensor Warfarin Sensitivity
Test and XT-8 System (K073720) | |
| | | | have an impact on the intended use
or the safety and effectiveness of the
device. |
| Power Requirements | Universal input, 90-264 Vac | 100-240 Vac, 50/60 Hz, 600 W | The power requirements of TruDx
2000 Imager and the eSensor XT-8
System are equivalent. Both systems
are 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 smaller
than the eSensor XT-8 System..
This difference in size does not have
an impact on the intended use or the
safety and effectiveness of the
device. |
| Weight | Approximately 25 lbs | 48.5 lbs | The TruDx 2000 Imager is lighter
than the eSensor XT-8 System.
This difference in weight does not
have an impact on the intended use
or the safety and effectiveness of the
device. |
| Specimen type | Genomic DNA obtained from
epithelial cells contained in human
saliva samples collected using the
Oragene Dx Device OGD-500. | Genomic DNA obtained from a
human whole blood sample. | The specimen type of the
TruDiagnosis System is different
from the specimen type of the
eSensor Warfarin Sensitivity Test
and XT-8 System.
This difference does not affect the
safety and effectiveness or intended
use of the device. |
| Number of SNPs | 3 | 3 | The TruDiagnosis System is identical
to the eSensor Warfarin Sensitivity
Test and XT-8 System. |
| Item | Subject Device | Predicate Device | Comparison |
| | TruDiagnosis® System | eSensor Warfarin Sensitivity
Test and XT-8 System (K073720) | |
| Input gDNA | 25 ng of genomic DNA (5 µL of 5
ng/µL genomic DNA sample) | 10 ng of genomic DNA (5 µL of 2
ng/µL genomic DNA sample) | The TruDiagnosis System is different
from the eSensor Warfarin
Sensitivity Test and XT-8 System. |
| DNA Extraction | Qiagen method. QIAAMP DNA Mini
Kit. | Manual ethanol extraction method | The TruDiagnosis System is different
from the eSensor Warfarin
Sensitivity Test and XT-8 System
Test.
This difference does not affect the
safety and effectiveness or intended
use of the device. |
| PCR Process | Closed amplicon. On-slide PCR
occurs in the same chamber as the
microarray. Asymmetric PCR is
performed on-slide to eliminate the
predicate's exonuclease step. | Open amplicon. Two steps occur
off cartridge: 1) tube-based PCR
and 2) exonuclease enzyme
treatment to generate single
stranded target DNA. | The PCR Process of the
TruDiagnosis System is different
from that of the eSensor Warfarin
Sensitivity Test and XT-8 System.
This difference does not affect the
safety and effectiveness or intended
use of the device. |
| Thermal Cycling Used | Yes | Yes | The TruDiagnosis System is identical
to the eSensor Warfarin Sensitivity
Test and XT-8 System. |
| Target of Detection | Single-nucleotide polymorphism | Single-nucleotide polymorphism | The TruDiagnosis System is identical
to the eSensor Warfarin Sensitivity
Test and XT-8 System. |
| Protocol Identification | Uses a barcode to store protocol and
production information | Uses an internal memory chip
(EEPROM) to store protocol and
production information. | The TruDiagnosis System is
equivalent to the eSensor Warfarin
Sensitivity Test and XT-8 System. |

Detailed Comparison of the Subject and Predicate Devices:

7

8

9

10

11

12

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.

13

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.

14

Limit of Detection Study by Sample Genotype

		Genotype				First Pass Calls						Final Resulta
Donor	2C9
*2	2C9
*3	VKORC1	DNA
Input
(ng)	#
Replicates
Tested	#
Correct
Calls	#
No
Calls	#
Incorrect
Calls	Correct
Call
Rateb	95%
LCB	#
Correct
Calls	#
No
Calls	#
Incorrect
Calls	Correct
Call
Rateb	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.

15

Limit of Detection Study by Loci

					First Pass Calls		Final Result a
DNA
Amount
(ng)	Loci	Total
Number
of Calls	#
Correct
Calls	#
No
Calls	#
Incorrect
Calls	Correct
Call
Rate b	95%
LCB	#
Correct
Calls	#
No
Calls	#
Incorrect
Calls	Correct
Call
Rate 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.

16

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.

17

| | Bi-directional Sequencing
Sample Genotype | | | |-------|----------------------------------------------|--- | 2C92 | 2C93 Samples
Tested | #
Correct
Calls | #
No
Calls | #
Incorrect
Calls | Correct
Call
Rate b | 95%
LCB | #
Correct
Calls | #
No
Calls | #
Incorrect
Calls | Correct
Call
Rate b | 95%
LCB |
| *1/*1 | *1/*1 | *1/*1 | *1/*1 | *1/*1 | *1/*1 | *1/*2 | *1/*1 | *1/*2 | *1/*1 | *1/*2 | *1/*1 | *1/*1 | *1/*3 | *1/*1 | *1/*3 | *1/*1 | *1/*3 | *2/*2 | *1/*1 | *2/*2 | *1/*1 | *2/*2 | *1/*1 | *1/*1 | *3/*3 | *1/*2 | *1/*3 | *1/*2 | *1/*3 | | Total | | First Pass Calls | | Final Result a | | | | | | |
-----|------------------------|-----------------------|------------------|-------------------------|---------------------------|------------|-----------------------|------------------|-------------------------|---------------------------|------------|
| VKORC1 | #
| AA | 58 | 57 | 1 | 0 | 98.3% | 92.1% | 58 | 0 | 0 | 100.0% | 95.0% |
| GA | 79 | 79 | 0 | 0 | 100.0% | 96.3% | 79 | 0 | 0 | 100.0% | 96.3% |
| GG | 41 | 41 | 0 | 0 | 100.0% | 93.0% | 41 | 0 | 0 | 100.0% | 93.0% |
| AA | 8 | 8 | 0 | 0 | 100.0% | 68.8% | 8 | 0 | 0 | 100.0% | 68.8% |
| GA | 21 | 21 | 0 | 0 | 100.0% | 86.7% | 21 | 0 | 0 | 100.0% | 86.7% |
| GG | 30 | 29 | 1 | 0 | 96.7% | 85.1% | 30 | 0 | 0 | 100.0% | 90.5% |
| AA | 8 | 7 | 1 | 0 | 87.5% | 52.9% | 8 | 0 | 0 | 100.0% | 68.8% |
| GA | 15 | 15 | 0 | 0 | 100.0% | 81.9% | 15 | 0 | 0 | 100.0% | 81.9% |
| GG | 25 | 25 | 0 | 0 | 100.0% | 88.7% | 25 | 0 | 0 | 100.0% | 88.7% |
| AA | 1 | 1 | 0 | 0 | 100.0% | 5.0% | 1 | 0 | 0 | 100.0% | 5.0% |
| GA | 6 | 5 | 0 | 1 c | 83.3% | 41.8% | 6 | 0 | 1 c | 83.3% | 41.8% |
| GG | 3 | 3 | 0 | 0 | 100.0% | 36.8% | 3 | 0 | 0 | 100.0% | 36.8% |
| GG | 1 | 1 | 0 | 0 | 100.0% | 5.0% | 1 | 0 | 0 | 100.0% | 5.0% |
| GA | 6 | 6 | 0 | 0 | 100.0% | 60.7% | 6 | 0 | 0 | 100.0% | 60.7% |
| GG | 1 | 1 | 0 | 0 | 100.0% | 5.0% | 1 | 0 | 0 | 100.0% | 5.0% |
| 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.

18

| Bi-directional Sequencing

Sample Genotype			First Pass Calls						Final Result a
Loci	Genotype	#
Samples
Tested	#
Correct
Calls	#
No
Calls	#
Incorrect
Calls	Correct
Call
Rate b	95%
LCB	#
Correct
Calls	#
No
Calls	#
Incorrect
Calls	Correct
Call
Rate 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.

19

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.

| Interference
Condition | Interference
Concentration | #
Samples
Tested | #
Correct
Calls | #
No
Calls | #
Incorrect
Calls | Correct
Call
Rate b | 95%
LCB | #
Correct
Calls | #
No
Calls | #
Incorrect
Calls | Correct
Call
Rate b | 95%
LCB |
|---------------------------|-------------------------------|------------------------|-----------------------|------------------|-------------------------|---------------------------|------------|-----------------------|------------------|-------------------------|---------------------------|------------|
| Control Total | N/A | 15 | 15 | 0 | 0 | 100.0% | 81.9% | 15 | 0 | 0 | 100.0% | 81.9% |
| Amylase Total | 311 - 341 U | 15 | 15 | 0 | 0 | 100.0% | 81.9% | 15 | 0 | 0 | 100.0% | 81.9% |
| Hemoglobin
Total | 21.2 - 22.1 mg/mL | 15 | 14 | 1 | 0 | 93.3% | 72.1% | 15 | 0 | 0 | 100.0% | 81.9% |
| IgA Total | 0.3 - 0.5 mg/mL | 15 | 15 | 0 | 0 | 100.0% | 81.9% | 15 | 0 | 0 | 100.0% | 81.9% |
| Protein Total | 2.1 - 2.7 mg/mL | 15 | 13 | 2 | 0 | 86.7% | 63.7% | 15 | 0 | 0 | 100.0% | 81.9% |

Endogenous Interference

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

20

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.

21

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 | #
Samples
Tested | #
Correct
Calls | #
No
Calls | #
Incorrect
Calls | Correct
Call
Rate b | 95%
LCB | #
Correct
Calls | #
No
Calls | #
Incorrect
Calls | Correct
Call
Rate b | 95%
LCB |
|--------------------|----------|--------|------------------------|-----------------------|------------------|-------------------------|---------------------------|-----------------|-----------------------|------------------|-------------------------|---------------------------|------------|
| First Pass Results | | | | | | | | Final Results a | | | | | |
| 1 | 1 | 2C92 | 90 | 78 | 12 | 0 | 86.7% | 79.3% | 90 | 0 | 0 | 100.0% | 96.7% |
| 1 | 1 | 2C93 | 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 | 2C92 | 90 | 82 | 8 | 0 | 91.1% | 84.5% | 90 | 0 | 0 | 100.0% | 96.7% |
| 1 | 2 | 2C93 | 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 | 2C92 | 90 | 71 | 19 | 0 | 78.9% | 70.6% | 90 | 0 | 0 | 100.0% | 96.7% |
| 2 | 3 | 2C93 | 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 | 2C92 | 90 | 80 | 10 | 0 | 88.9% | 81.9% | 90 | 0 | 0 | 100.0% | 96.7% |
| 3 | 4 | 2C93 | 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 | 2C92 | 360 | 311 | 49 | 0 | 86.4% | 83.1% | 360 | 0 | 0 | 100.0% | 99.2% |
| Total | Loci | 2C93 | 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	#
Samples
Tested	#
Correct
Calls	#
No
Calls	#
Incorrect
Calls	Correct
Call
Rate b	95%
LCB	#
Correct
Calls	#
No
Calls	#
Incorrect
Calls	Correct
Call
Rate b	95%
LCB
#1
5007/5008	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%
#2
0011/0008	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%
#3
8005/8004	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%
#4
6010/0015	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%
All
Lots	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.

25

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).