The 23andMe Personal Genome Service (PGS) is a qualitative genotyping assessment system applied to genomic DNA isolated from human saliva collected using the Oragene Dx OGD-500.001 to simultaneously detect, report, and interpret genetic variants in a broad multigene test. The assessment system is intended to enable users to access information about their genetics that could aid discussions with a healthcare professional. The 23andMe Pharmacogenetic Reports are indicated for reporting of the following variants:

Gene: CYP2C19 Variant(s): *2, *3, *17
Gene: CYP2C9 Variant(s): *2, *3, *5, *6, rs7089580
Gene: CYP3A5 Variant(s): *3
Gene: UGT1A1 Variant(s): *6, *28
Gene: DPYD Variant(s):*2A, rs67376798
Gene: TPMT Variant(s): *2, *3C
Gene: SLC01B1 Variant(s): *5
Gene: CYP2D6 Variant(s): *2, *3, *4, *5, *6, *7, *8, *9, *10, *11, *15, *17, *20, *29, *35, *40, *41

This report is for over-the-counter use by adults over the age of 18 and provides genetic information to inform discussions with a healthcare professional about metabolism of therapeutics.

The 23andMe Personal Genome Service pharmacogenetic reports for CYP2C9, CYP3A5, UGT1A1, DPYD, TPMT, SLC01B1 and CYP2D6 describe if a person has variants associated with metabolism of some therapeutics, but does not describe if a person will or will not respond to a particular therapeutic, and does not describe the association between detected variants and any specific therapeutic.

23andMe Personal Genome Service pharmacogenetic reports for CYP2C19 describes if a person has variants associated with metabolism of some therapeutics and provides interpretive drug information regarding the potential effect of the identified metabolizer phenotype on citalopram and clopidogrel therapy.

The PGS Pharmacogenetic Reports are not a substitute for visits to a healthcare professional. The information provided by this report should not be used to start, stop, or change any course of treatment.

Device Description

The 23andMe Personal Genome Service (PGS) is an over-the-counter (direct-to-consumer), DNA testing service that provides information and tools for consumers to learn about and explore their DNA.

The PGS is a currently marketed, non-invasive genetic information service that combines qualitative genotyping data covering genetic ancestry, traits, and certain heritable health conditions from a single multiplex assay with descriptive information derived from peer reviewed, published genetic research studies.

Customer saliva specimens are self-collected using the Oragene-Dx® Device manufactured by DNA Genotek, Inc. cleared by FDA for use with the PGS device (K141410, DEN140044, DEN160026, DEN170046, DEN180028, and K182784), which consists of a sealable collection tube containing a stabilizing buffer solution. Once the sample is collected, it is shipped to a Clinical Laboratory Improvement Amendments (CLIA) certified laboratory for testing.

DNA is isolated from the saliva and tested in a multiplex assay using a customized genotyping beadchip, reagents and instrumentation manufactured by Illumina. The device simultaneously tests for more than 600,000 variants, including those reported under the previously authorized PGS test indications.

The raw data is generated using Illumina GenomeStudio software, and then sent to 23andMe for analysis and interpretation. The raw data received is analyzed using 23andMe's proprietary Coregen software, where a genotype is determined for each tested SNP. The results for certain of these SNPs are used to generate personalized reports for the customer that provide information about the detected genotype.

Personalized reports are generated for each user that provide results of the testing performed. These reports tell the user which variant(s) has/have been detected in their sample and provide information on metabolizer or transporter profile associated with the variant(s). If no variant was detected, that information is also provided. The personalized reports are designed to present scientific concepts to users in an easy-to-understand format. The reports provide scientifically valid information about the metabolizer or transporter phenotype associated with the presence of a particular variant, or a combination of variants. In the pharmacogenetic report for CYP2C19, information regarding interpretive drug information to certain medications will be provided to the user in a medication "mini report", which is accessed via a link in the pharmacogenetic report for CYP2C19. The reports are designed to help users understand the meaning of their results and any appropriate actions that may be taken based on their results.

The novel components in this traditional 510(k) submission are to provide interpretive drug information to two specific medications (citalopram and clopidogrel), and to remove the limitation language requiring confirmatory testing in the 23andMe pharmacogenetic report for CYP2C19. Pharmacogenetic reports for the other genes authorized in DEN180028 will not be modified to include interpretive drug information, or remove the confirmatory testing limitation.

AI/ML Overview

Here's an analysis of the acceptance criteria and the supporting study for the 23andMe Personal Genome Service (PGS) Pharmacogenetic Reports, specifically focusing on the CYP2C19 gene, as described in the provided document:

1. Table of Acceptance Criteria and Reported Device Performance

The document focuses on the analytic performance of the genotyping assay for CYP2C19 variants (*2, *3, *17). The primary acceptance criteria provided relate to accuracy (method comparison) and precision (reproducibility).

Acceptance Criteria Category	Specific Acceptance Criteria	Reported Device Performance (CYP2C19 variants: 2, 3, *17)
Method Comparison (Accuracy)	>99% agreement with the source of truth (PPA and NPA both >99%)	100% overall agreement for all genotypes of each of the three variants, with Sanger sequencing as the source of truth. 100% concordance to comparator source of truth for blinded studies (Coriell samples and East Asian saliva samples), achieving >99% PPA and >99% NPA.
Precision (Reproducibility)	≥ 99% correct calls	>99% correct genotype calls for all samples across multiple days, operator teams, instruments, and reagent lots at two independent laboratory sites (initial study). 100% correct genotype calls for all samples across multiple days, operator teams, instruments, and reagent lots at two independent laboratory sites (supplemental saliva study). 100% reproducibility and 100% repeatability for intended use (saliva) samples.
Minimum DNA Input (MDI)	≥ 95% of samples yielded the correct call at the lowest DNA concentration	100% concordant test results/correct genotype calls for all samples and all reagent lots tested at sample DNA concentrations of 5, 15, and 50 ng/μL. Passed acceptance criteria at a sample DNA concentration of 5 ng/μL for both initial and supplemental studies.

2. Sample Sizes and Data Provenance for the Test Set

Method Comparison (Accuracy) & Precision (Reproducibility) Test Sets:
- "Blinded Study 1" (single lab site, single day): Five 96-well plates of "incoming saliva samples." A specific numerical count of samples is not given, but 5 plates x 96 wells/plate = 480 samples.
- Coriell Reference Samples: Genomic DNA for the three CYP2C19 variants of interest (*2, *3, *17) was obtained from the Coriell Institute for Medical Research. The exact number of Coriell samples used is not specified.
- "Blinded Study 2" (East Asian ancestry): Intended use (saliva) samples "randomly selected in an unbiased manner from the 23andMe biobank based on their East Asian genetic ancestry." Numbers not specifically given. The purpose was to increase the likelihood of rare *3 allele combinations.
- Supplemental Precision Study (saliva): Intended use (saliva) samples selected from the 23andMe customer biobank based on their "putative genotype." Numbers not specifically given but tested over 3 days, with 3 lots of reagents, by unique operator teams, using 3 different serial numbers of each of 2 instruments (Tecan and iScan), at each of 2 laboratory sites.
- Minimum DNA Input (MDI) Test Set:
  - DNA samples obtained from Coriell and the 23andMe biobank. Exact numbers not specified, but tested at 3 different DNA concentrations with 3 reagent lots.
  - Supplemental MDI Study (saliva): Intended use (saliva) samples selected from the 23andMe customer database based on their "putative genotype." Numbers not specified, but each sample was diluted to 3 different DNA concentrations and genotyped with 3 reagent lots.
Data Provenance:
- Retrospective: Samples from the 23andMe biobank and customer database are retrospective, representing previously collected data.
- Prospective (Implicit): "Incoming saliva samples" in the first blinded study could imply prospective collection for that specific study's initiation if they were fresh samples received for routine processing on that day.
- External Reference: Coriell Institute for Medical Research samples are an external, well-characterized source.
- Country of Origin: Not explicitly stated for all samples, but the 23andMe customer base is likely predominantly from the US and other countries where they operate.

3. Number of Experts and Qualifications for Ground Truth Establishment (Test Set)

This device is a genotyping system, and the ground truth for its analytical performance is established through genomic sequencing, not expert interpretation of phenotypic data or images.

No human experts were used to establish the ground truth in the traditional sense of clinical interpretation.
The "experts" in this context are the robust and established methods of bi-directional Sanger sequencing and Coriell Institute's characterization of their reference materials. These methods are considered the "source of truth" for genetic sequences and genotypes.

4. Adjudication Method for the Test Set

No human adjudication method (like 2+1, 3+1) was used as the ground truth is based on objective genetic sequencing data.
The comparison involved directly comparing the device's genotyping results against the known sequences/genotypes derived from Sanger sequencing or Coriell's reference data. Discrepancies would likely lead to re-sequencing or further investigation to ascertain the true genotype.

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

No, a multi-reader multi-case (MRMC) comparative effectiveness study was not done.
This device is an automated genotyping system, not one that involves human "readers" interpreting output in a diagnostic setting in the way an imaging AI might. Its primary function is to accurately identify genetic variants. The "interpretive drug information" is a downstream output of the accurate genotyping, not something itself interpreted by multiple human readers in a study of this type.

6. Standalone Performance Study (Algorithm Only)

Yes, the performance studies described (Method Comparison, Precision, Minimum DNA Input) are all standalone performance studies of the genotyping algorithm and associated laboratory process.
The studies evaluate the accuracy and reliability of the device itself (the genotyping platform, reagents, and analysis software) in determining genetic variants, separate from how a human might interpret the final report.

7. Type of Ground Truth Used

Expert Consensus (Genomic): The ground truth for the analytical performance studies was established using:
- Bi-directional Sanger Sequencing: A gold standard method for DNA sequencing, considered highly accurate for confirming specific genetic variants.
- Reference Samples from Coriell Institute for Medical Research: These are well-characterized genomic DNA samples with established and verified genotypes, essentially an expert-characterized reference.

8. Sample Size for the Training Set

The document does not explicitly state a separate "training set" for the genotyping algorithm. Genetic genotyping algorithms often rely on established biochemical principles and mapping to known reference genomes rather than machine learning on a large, labeled training dataset of raw signals to call genotypes.
However, the "23andMe database" is mentioned, with variant and allele frequencies from "8,004,302 customers" and "16,008,604 alleles." This massive dataset, along with public databases like gnomAD, would likely be used in the development and refinement of allele frequency estimation and potentially for algorithm fine-tuning or quality control, but not as a distinct "training set" in the typical AI sense for classifying novel inputs.

9. How the Ground Truth for the Training Set was Established

Given that a distinct "training set" as understood in machine learning is not explicitly detailed, the ground truth establishment for algorithm development would implicitly rely on the same principles as the test set: established genetic sequencing methodologies (like Sanger sequencing) and comprehensive genetic reference databases (like the human reference genome and population-specific variant databases). These resources allow for the accurate mapping and identification of genetic variants within the raw genotyping data.

Summary

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left, there is a symbol representing the Department of Health & Human Services - USA. To the right of the symbol, there is the FDA logo in blue, with the words "U.S. FOOD & DRUG" stacked above the word "ADMINISTRATION".

August 17, 2020

23andMe, Inc. Marianna Frendo Program Manager, Regulatory Affairs 223 N Mathilda Ave Sunnyvale, CA 94086

Re: K193492

Trade/Device Name: 23andMe Personal Genome Service (PGS) Pharmacogenetic Reports Regulation Number: 21 CFR 862.3364 Regulation Name: Pharmacogenetic Assessment System Regulatory Class: Class II Product Code: ODJ Dated: July 17, 2020 Received: July 17, 2020

Dear Marianna Frendo:

We have reviewed vour 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/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); 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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). 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 (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-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. 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) K193492

Device Name

23andMe® Personal Genome Service® (PGS) Pharmacogenetic Reports pharmacogenetic report for CYP2C19, CYP3A5, UGT1A1, DPYD, TPMT, SLC01B1, CYP2D6

Indications for Use (Describe)

Gene: CYP2C19 Variant(s): *2, *3, *17 Gene: CYP2C9 Variant(s): *2, *3, *5, *6, rs7089580 Gene: CYP3A5 Variant(s): *3 Gene: UGT1A1 Variant(s): *6, *28 Gene: DPYD Variant(s):*2A, rs67376798 Gene: TPMT Variant(s): *2, *3C Gene: SLC01B1 Variant(s): *5 Gene: CYP2D6 Variant(s): *2, *3, *4, *5, *6, *7, *8, *9, *10, *11, *15, *17, *20, *29, *35, *40, *41

This report is for over-the-counter use by adults over the age of 18 and provides genetic inform discussions with a healthcare professional about metabolism of therapeutics.

The 23andMe Personal Genome Service pharmacogenetic reports for CYP2C9, CYP3A5, UGT1A1, DPYD, TPMT, SLCO1B1 and CYP2D6 describe if a person has variants associated with metabolism of some therapeutics, but does not describe if a person will or will not respond to a particular therapeutic, and does not describe the association between detected variants and any specific therapeutic.

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)

[X] Over-The-Counter Use (21 CFR 801 Subpart C)

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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."

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SECTION 5 - 510(k) SUMMARY

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of Safe Medical Devices Act of 1990 and 21 CFR 807.92.

The assigned 510(k) number is:

Submitter / Company

Lisa Charter Director, Regulatory Affairs 650-933-9455 LCharter@23andMe.com Establishment Registration Number: 3007699459 Owner Operator Number: 10029404

Company Contact

Marianna Frendo Program Manager, Regulatory Affairs 650-933-9481 ext. 7281 MFrendo@23andMe.com

Date Prepared

14Aug2020

5.1. REGULATORY INFORMATION

Table 5.1 Proposed New Device

Type of Submission:	Traditional 510(k)
Common/Usual Name:	pharmacogenetic reports for CYP2C19, CYP2C9, CYP3A5,UGT1A1, DPYD, TPMT, SLCO1B1, CYP2D6
Trade/Proprietary Name:	23andMe Personal Genome Service (PGS) PharmacogeneticReports

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Regulation Description:	A pharmacogenetic assessment system is a qualitative in vitromolecular diagnostic system intended to detect nucleic acidvariants isolated from human specimens for the purpose ofassessing the presence of genetic variants that impact themetabolism, exposure, response, risk of adverse events,dosing, or mechanisms of prescription or over-the-countermedications. The intended use of the device must not includean indication for use in supporting or sustaining human life,being of substantial importance in preventing impairment ofhuman health, or presenting a potential, unreasonable risk ofillness or injury.
Regulation Number:	21 CFR 862.3364
Product Code:	QDJ
Class:	Class II
Predicate Device:	23andMe PGS Pharmacogenetic Reports authorized onOctober 31, 2018 under DEN180028

5.2. INDICATIONS FOR USE

Gene	Variant(s)
CYP2C19	2, 3, *17
CYP2C9	2, 3, 5, 6, rs7089580
CYP3A5	*3
UGT1A1	6, 28
DPYD	*2A, rs67376798

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TPMT	2, 3C
SLCO1B1	*5
CYP2D6	2, 3, 4, 5, 6, 7, 8, 9, 10,11, 15, 17, 20, 29, 35, 40,*41

This report is for over-the-counter use by adults over the age of 18 and provides genetic information to inform discussions with a healthcare professional about metabolism of therapeutics.

5.3. SUBSTANTIALLY EQUIVALENT PREDICATE DEVICE

The components of the assay are unchanged from the de novo authorization for the predicate device. These components include the saliva collection kit, the reagents and BeadChip, the instrumentation, the software, the test processes, and procedures.

The purpose of this traditional 510(k) submission is to modify the labeling of the 23andMe Personal Genome Service (PGS) Pharmacogenetic Reports, pharmacogenetic report for CYP2C19, to add one new indication describing interpretive drug information to two specific medications (citalopram and clopidogrel), as well as to remove the limitation language requiring confirmatory testing. Interpretive drug information will be provided to users in the pharmacogenetic report for CYP2C19, and will be accessed through medication specific links in the pharmacogenetic report for CYP2C19. Pharmacogenetic reports for the other genes authorized in DEN180028 will not be modified to include interpretive druq information, or remove the confirmatory testing limitation statement.

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The pharmacogenetic report for CYP2C19 results, scientific details, and FAQs were designed and developed in the same format as the predicate device, and conform to the requirements described in DEN180028.

Specifically, the pharmacogenetic report for CYP2C19 is intended for over-the-counter, direct-toconsumer use without prescription or physician order. In both the predicate device and the proposed modified device, all customers are advised to share their results with their healthcare provider regardless of their results. Customers with altered function of the CYP2C19 enzyme are strongly advised to share their results with their healthcare provider. The proposed revised labeling of the modified device includes certain healthcare provider limitations, as required. The Package Insert has been revised to incorporate information pursuant to agreements specific to this submission such modifications necessary to remove the confirmatory testing requirement for CYP2C19, and to address the inclusion of the pharmacogenetic association information for CYP2C19, consistent with the product classification under 21 CFR 862.3364. The proposed modifications are based on the additional test system performance data submitted in this 510(k).

Warnings, precautions, and limitations for CYP2C9, CYP3A5, UGT1A1, DPYD, TPMT, SLCO1B1. CYP2D6

. Do not use your results to start, stop or change any course of treatment.
. Results from this test should not be used to make medical decisions. Results should be confirmed by an independent genetic test that is prescribed by your attending physician before taking any medical action.
. This test does not provide information on associations between specific DNA variants and any specific therapeutic.
. This test does not diagnose any health conditions, predict drug response, provide medical advice, or determine whether a medication is indicated for the user.
. This report does not determine if a person will not respond to a particular therapeutic.
This test does not detect all genetic variants related to drug metabolism. The ● absence of a variant tested does not rule out the presence of other genetic variants that may be related to drug metabolism.
This test is not a substitute for visits to a healthcare professional. You should consult with a healthcare professional if you have any questions or concerns about your results.
This test may not be able to determine a result for all variants analyzed.
. Different companies offering genetic testing may be measuring different genetic variants for drug metabolism, so you may get different results from a different test.
As with every test the possibility for an incorrect result exists. Speak to your personal ● healthcare professional or a genetic counselor if your results are unexpected.

Warning, precautions, and limitations specific for only CYP2C19

Do not use your results to start, stop or change any course of treatment. .
This test does not diagnose any health conditions, provide medical advice, or determine whether a medication is indicated for the user.

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This test provides interpretive drug information on certain therapeutics.
. This report does not determine if a person will not respond to a particular therapeutic.
. This test does not detect all genetic variants related to drug metabolism. The absence of a variant tested does not rule out the presence of other genetic variants that may be related to drug metabolism.
. This test is not a substitute for visits to a healthcare professional. You should consult with a healthcare professional if you have any questions or concerns about your results.
This test may not be able to determine a result for all variants analyzed.
This test does not provide interpretive drug information for the CYP2C19 *3/*17 ● heterozygous genotype. In addition, results for this genotype should be confirmed by an independent genetic test that is prescribed by your attending physician before taking any medical action.
. Different companies offering genetic testing may be measuring different genetic variants for drug metabolism, so you may get different results from a different test.
. As with every test the possibility for an incorrect result exists. Speak to your personal healthcare professional or a genetic counselor if your results are unexpected.

This submission proposes an intended use for the pharmacogenetics report for CYP2C19 that does not raise new questions of safety and effectiveness and is supported by performance data collected for this purpose, or similar. As such, the modified 23andMe Personal Genome Service Pharmacogenetic Reports is substantially equivalent to the predicate device authorized under DEN180028.

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5.4. DEVICE DESCRIPTION

The 23andMe Personal Genome Service (PGS) is an over-the-counter (direct-to-consumer), DNA testing service that provides information and tools for consumers to learn about and explore their DNA.

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Engineering drawings, schematics, etc. of the 23andMe Personal Genome Service (PGS) Pharmacogenetic Reports are not applicable to this device.

5.5. TECHNOLOGICAL CHARACTERISTICS

Test Type: Qualitative genetic test for single nucleotide polymorphism detection.

Sample Type: Genomic DNA obtained from a human saliva sample.

Target of detection: Single-nucleotide polymorphism.

DNA extraction: Automated and manual methods.

Gene: CYP2C19, CYP2C9, CYP3A5, UGT1A1, DPYD, TPMT, SLCO1B1, CYP2D6

Variants: *2, *3, *17 variants in the CYP2C19 gene, *2, *3, *6, rs7089580 in the CYP2C9 gene, *3 in the CYP3A5 gene, *6, *28 in the UGT1A1 gene, *2A, rs67376798 in the DPYD gene, *2, *3C in the TPMT gene, *5 in the SLCO1B1 gene, *2, *3, *4, *5, *6, *7, *8, *9, *10, *11, *15, *17, *20, *29, *35, *40, *41 in the CYP2D6 gene

SNPs: *2(rs4244285), *3(rs4986893), *17(rs12248560)

Genotyping principle: The DNA is fragmented and captured on a bead array by hybridization to immobilized SNP-specific primers, followed by extension with hapten-labeled nucleotides. The primers hybridize adjacent to the SNPs and are extended with a single nucleotide corresponding to the SNP allele. The incorporated hapten-modified nucleotides are detected by adding fluorescently labeled antibodies in several steps to amplify the signals. Data analysis is performed using scatter plots.

Instrument: Illumina iScan and GenomeStudio system.

Assay results: The genotype content is separated, and then integrated into predefined report templates specific for each condition associated with each genotype.

	PredicatePharmacogenetic ReportsDEN180028	Proposed modifiedindication for thepharmacogenetic reportfor CYP2C19	SimilaritiesandDifferences
Indicationsfor Use	The 23andMe PersonalGenome Service (PGS) is aqualitative genotypingassessment system appliedto genomic DNA isolatedfrom human saliva tosimultaneously detect,report, and interpret geneticvariants in a broadmultigene test. Theassessment system isintended to enable users to	The 23andMe PersonalGenome Service (PGS) is aqualitativegenotypingassessment system appliedto genomic DNA isolatedfrom human saliva collectedusing the Oragene Dx OGD-500.001 to simultaneouslydetect, report, and interpretgenetic variants in a broadmultigene test. Theassessment system is	Modificationof theindication toaddinterpretivedruginformationfor CPY2C19and forremoval of therequirementfor

	Table 5.2 Substantial Equivalence

access information abouttheir genetics that could aiddiscussions with ahealthcare professional.Gene: CYP2C19Variant(s): 2, 3, 17Gene: CYP2C9Variant(s): 2, 3, 5, 6,rs7089580Gene: CYP3A5Variant(s): 3Gene: UGT1A1Variant(s): 6, 28Gene: DPYDVariant(s):2A, rs67376798Gene: TPMTVariant(s): 2, 3CGene: SLCO1B1Variant(s): 5Gene: CYP2D6Variant(s): 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 15, 17,20, 29, 35, 40, *41This report is for over-the-counter use by adults overthe age of 18 and providesgenetic information to informdiscussions with ahealthcare professionalabout metabolism oftherapeutics.This report describes if aperson has variantsassociated with metabolismof some therapeutics, butdoes not describe if aperson will or will notrespond to a particulartherapeutic, and does notdescribe the association	intended to enable users toaccess information abouttheir genetics that could aiddiscussions with ahealthcare professional. The23andMe PharmacogeneticReports are indicated forreporting of the followingvariants:Gene: CYP2C19Variant(s): 2, 3, 17Gene: CYP2C9Variant(s): 2, 3, 5, 6,rs7089580Gene: CYP3A5Variant(s): 3Gene: UGT1A1Variant(s): 6, 28Gene: DPYDVariant(s):2A, rs67376798Gene: TPMTVariant(s): 2, 3CGene: SLCO1B1Variant(s): 5Gene: CYP2D6Variant(s): 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 15, 17,20, 29, 35, 40, *41This report is for over-the-counter use by adults overthe age of 18 and providesgenetic information to informdiscussions with ahealthcare professionalabout metabolism oftherapeutics.The 23andMe PersonalGenome ServicePharmacogenetic Reportsfor CYP2C9, CYP3A5,	confirmatorytesting. Newclaims arelimited to theCYP2C19variant.

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	between detected variantsand any specific therapeutic.The PGS PharmacogeneticReports are not a substitutefor visits to a healthcareprofessional. Theinformation provided by thisreport should not be used tostart, stop, or change anycourse of treatment.	UGT1A1, DPYD, TPMT,SLCO1B1 and CYP2D6describe if a person hasvariants associated withmetabolism of sometherapeutics, but does notdescribe if a person will orwill not respond to aparticular therapeutic, anddoes not describe theassociation betweendetected variants and anyspecific therapeutic.23andMe Personal GenomeService pharmacogeneticreports for CYP2C19describes if a person hasvariants associated withmetabolism of sometherapeutics and providesinterpretive drug informationregarding the potential effectof the identified metabolizerphenotype on citalopram andclopidogrel therapy.The PGS PharmacogeneticReports are not a substitutefor visits to a healthcareprofessional. The informationprovided by this reportshould not be used to start,stop, or change any courseof treatment.
CollectionKit	Oragene·Dx® salivacollection device (OGD-500.001)K141410	Oragene·Dx® salivacollection device (OGD-500.001)K141410	Same
Reagents	Illumina Infinium HTS AssayReagents	Illumina Infinium HTS ExtraAssay Reagents	Samereagents.Different.
			packaging.
BeadChip	Illumina Global ScreeningArray customized for thePGS. The chip is designedto detect specific singlenucleotide polymorphisms(SNPs) as well as othergenetic variants; all markersrefer to specific positions inthe National Center forBiotechnology Information(NCBI) reference humangenome.	Illumina Global ScreeningArray customized for thePGS. The chip is designedto detect specific singlenucleotide polymorphisms(SNPs) as well as othergenetic variants; all markersrefer to specific positions inthe National Center forBiotechnology Information(NCBI) reference humangenome.	Same.
Beadpool	Customized for 23andMe	Customized for 23andMe	Same
Instruments	Tecan EvoIllumina iScan	Tecan EvoIllumina iScan	Same
Software	Genome StudioCoregen	Genome StudioCoregen	Same
SampleMatrix	Saliva	Saliva	Same
MethodComparison	>99% PPA and NPA for allgenotypes	>99% PPA and NPA for allgenotypes	Same

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5.6. PERFORMANCE TESTING SUMMARY

The purpose of this submission is to modify the 23andMe Personal Genome Service (PGS) Pharmacogenetic Reports as follows:

1. Remove confirmatory testing requirement in the pharmacogenetic report for CYP2C19
1. Provide interpretive drug information for certain therapeutics, accessible through the pharmacogenetic report for CYP2C19

In addition to the analytical data previously reviewed during DEN180028, the Company generated additional data, and performed new method comparison, and minimum DNA input studies to assess the performance of the 23andMe Personal Genome Service (PGS) pharmacogenetic reports for CYP2C19, in support of the confirmatory testing requirement and clearance.

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5.6.1 Method Comparison (Accuracy)

23andMe performed a method comparison study using sequencing as the comparator to assess the accuracy of the assay. Results of the test were compared with sequencing results for samples with known *2 variant status, known *3 variant status, and known *17 variant status, The method comparison study vielded 100% overall agreement for all genotypes of each of the three variants in the CYP2C19 gene (*2 (rs4244285); *3 (rs4986893); *17 (rs12248560)) for all samples compared to Sanger sequencing. Therefore, the study passed the acceptance criteria of >99% agreement, with PPA and NPA both >99%. The method comparison study showed that the assay is comparable to bi-directional Sanger sequencing for all three of the CYP2C19 gene variants: *2 (rs4244285); *3 (rs4986893); *17 (rs12248560).

To mitigate the risk for false positive results potentially influenced by selecting samples with genotypes previously determined by the assay, two method comparison studies were performed utilizing a blinded sample selection method.

In the first blinded study, from a single laboratory site on a single day, incoming saliva samples were extracted, normalized, and aliquoted onto a 96 well plate, according to laboratory SOPs. Five 96-well plates from that day were sent for Sanger sequencing at an independent laboratory. Upon completion of genotyping, results were compared to sequencing results to calculate positive percent agreement (PPA) and negative percent agreement (NPA), with the bi-directional Sanger sequencing results considered as the source of truth.

Additionally, well characterized reference samples were obtained from the Coriell Institute for Medical Research, which provided genomic DNA for the three CYP2C19 variants of interest (*2. *3, *17). Genotyping results were compared to the reference results, with the reference results considered as the source of truth.

The genotyping data demonstrated 100% concordance to the comparator source of truth, achieving the pre-defined acceptance criteria of greater than 99% PPA and greater than 99% NPA.

In the second blinded study, intended use (saliva) samples were randomly selected in an unbiased manner from the 23andMe biobank based on their East Asian genetic ancestry, in order to increase the likelihood of obtaining rare *3 allele combinations. Upon completion of genotyping, results were compared to sequencing results to calculate positive percent agreement (PPA) and negative percent agreement (NPA), with the bi-directional Sanger sequencing results considered as the source of truth.

The genotyping data demonstrated 100% concordance to Sanger sequencing, achieving the predefined acceptance criteria of greater than 99% PPA and greater than 99% NPA.

5.6.2 Precision (Reproducibility)

23andMe performed a precision study to evaluate the following precision parameters of the assay: intra-assay, operator-to-operator, instrument, and reagent lot-to-lot. for the following three variants in the CYP2C19 gene *2 (rs4244285), *3 (rs41291556), *17

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(rs12248560). DNA samples were obtained from Coriell based upon their listed genotypes, or obtained from the 23andMe biobank. The precision study vielded >99% correct genotype calls for all samples across multiple days, operator teams, instruments, and reagent lots, at two (2) independent laboratory sites. The study passed the acceptance criteria of at least 99% correct calls, the study had >99% reproducibility and >99% repeatability.

In order to ensure that the assay accurately reflected the reproducibility of intended use (saliva) samples, a supplemental precision study was performed utilizing saliva samples. Intended use (saliva) samples were selected from the 23andMe customer biobank based on their putative genotype. These samples were genotyped by the 23andMe BeadChip assay in a blinded fashion over 3 days, with 3 lots of reagents, by a unique operator team per day, using 3 different serial numbers of each of 2 instruments (Tecan and iScan), at each of 2 laboratory sites. To confirm the BeadChip genotype, each sample was also sequenced by bi-directional Sanger sequencing. BeadChip genotypes were compared with sequenced genotypes to determine the rates of correct BeadChip genotype calls. The acceptance criteria is a minimum of 99% correct qenotype calls at each of two laboratory sites.

Results of this supplemental precision study vielded 100% correct genotype calls for all samples across multiple days, operator teams, instruments, and reagent lots at two independent laboratory sites. Therefore, the study passed the acceptance criteria of at least 99% correct calls. There was no variation between any study conditions or any replicates for a qiven sample. Intended use samples representing *3 CYP2C19 AA obtained in the original precision study (DEN180028) also yielded 100% correct genotype calls for all samples across multiple days, operator teams, instruments, and reagent lots at two independent laboratory sites.

The results of this Precision study demonstrate 100% reproducibility and 100% repeatability on intended use (saliva) samples for CYP2C19.

5.6.3 Minimum DNA Input (MDI)

23andMe performed a minimum DNA input study to determine the lowest concentration of DNA that is necessary for successful assignment of the correct genotypes for three variants in the CYP2C19 gene: *2 (rs4244285), *3 (rs41291556), *17 (rs12248560). DNA samples obtained from Coriell based upon their listed genotypes, or obtained from the 23andMe biobank, were diluted to 3 different DNA concentrations using 3 lots of reagents. Results of this MDI study yielded 100% concordant test results correct genotype calls for all samples and all reagent lots tested at sample DNA concentrations of 5, 15, and 50 ng/μL. Therefore, the study passed the acceptance criteria at a sample DNA concentration of 5 ng/μL.

In order to ensure that the assay accurately reflected the performance of intended use (saliva) samples at various DNA input levels, a supplemental minimum DNA input study was performed utilizing saliva samples. Intended use (saliva) samples were selected from the 23andMe customer database based on their putative genotype. The goal of the study was to determine the lowest concentration of DNA that is necessary for successful assignment of the correct

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genotypes. Each sample was diluted to 3 different DNA concentrations and genotyped in a blinded fashion using 3 lots of reagents. The MDI requirement is defined as the lowest DNA concentration at which at least 95% of samples yielded the correct call.

Results of this MDI study yielded 100% correct genotype calls for all samples and all reagent lots tested at sample DNA concentrations of 5, 15, and 50 ng/μL. Therefore, the study passed the acceptance criteria at a sample DNA concentration of 5 ng/μL. Intended use samples representing *3 CYP2C19 AA obtained in the original MDI study (DEN180028) also yielded the correct genotype calls for all samples and all reagent lots tested at all DNA concentrations, and therefore met the acceptance criteria at a sample DNA concentration of 5 ng/yL.

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Table 5.3 Clinical Performance Variant frequencies from 23andMe database and other public database

This table includes data that has been updated as of 14 March 2019, subsequent to the authorization of DEN180028.

Allele frequencies from gnomAD database were obtained from https://gnomad.broadinstitute.org/ on 14Mar2019.

Variant	Percent of 23andMecustomers with varianta	Allele frequencies among23andMe customersa	Allele frequencies from gnomAD
*2	n=8,004,302 customers	n=16,008,604 alleles	n=192,906 alleles
(rs4244285:A)	European: 27.09%	European: 14.62%	European (Non-Finnish): 14.68%
	African American: 31.61%	African American: 17.34%	African: 17.76%
	Ashkenazi Jewish: 24.75%	Ashkenazi Jewish: 13.27%	Ashkenazi Jewish: 13.20%
	East Asian: 51.87%	East Asian: 30.65%	East Asian: 30.75%
	Hispanic or Latino: 24.67%	Hispanic or Latino: 13.24%	Latino: 10.12%
	South Asian: 55.54%	South Asian: 33.62%	South Asian: 32.40%
	Middle Eastern: 21.00%	Middle Eastern: 11.19%	European (Finnish): 17.50%
	Other: 33.74%	Other: 18.71%	Otherb: 15.95%
	Overall: 28.15%	Overall: 15.33%	Overall: 17.49%
*3	n=8,004,302 customers	n=16,008,604 alleles	n=282,504 alleles
(rs4986893:A)	European: 0.04%	European: 0.02%	European (Non-Finnish): 0.02635%
	African American: 0.22%	African American: 0.11%	African: 0.03611%
	Ashkenazi Jewish: <0.01%	Ashkenazi Jewish: <0.01%	Ashkenazi Jewish: 0.000%
	East Asian: 12.49%	East Asian: 6.50%	East Asian: 6.261%
	Hispanic or Latino: 0.28%	Hispanic or Latino: 0.14%	Latino: 0.01981%
	South Asian: 0.67%	South Asian: 0.34%	South Asian: 0.3986%
	Middle Eastern: 0.24%	Middle Eastern: 0.12%	European (Finnish): 0.007976%
	Other: 3.50%	Other: 1.77%	Othera: 0.2357%
	Overall: 0.61%	Overall: 0.32%	Overall: 0.5097%
*17	n=8,004,302 customers	n=16,008,604 alleles	n=31,250 alleles
(rs12248560:T)	European: 38.74%	European: 21.76%	European (Non-Finnish): 23.14%
	African American: 38.79%	African American: 21.78%	African: 20.92%
	Ashkenazi Jewish: 38.41%	Ashkenazi Jewish: 21.57%	Ashkenazi Jewish: 18.97%
	East Asian: 1.72%	East Asian: 0.86%	East Asian: 0.7051%
	Hispanic or Latino: 29.82%	Hispanic or Latino: 16.30%	Latino: 10.07%
	South Asian: 30.73%	South Asian: 16.96%	South Asian: n/a
	Middle Eastern: 37.72%	Middle Eastern: 21.18%	European (Finnish): 18.84%
	Other: 29.77%	Other: 16.24%	Othera: 22.55%
	Overall: 36.05%	Overall: 20.17%	Overall: 20.52%

a Small changes to observed allele and variant frequencies are expected to occur as the database grows.

Frequencies are truncated at a minimum frequency if the number of carriers is fewer than 5.

ኮ Individuals were classified as "Other" by gnomAD if they did not unambiguously cluster with the major populations (i.e. EUR, NFE, AFR, SAS, EAS and AMR) in a principal component analysis.

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5.7. DISCUSSION

The modified 23andMe Personal Genome Service (PGS) Pharmacogenetic Reports has an anticipated intended use under DEN180028 subject to the additional analytical data, is not technologically different than the predicate, and presents no new issues of safety or effectiveness when compared to the previously authorized predicate device (DEN180028). Specifically, this submission is for the removal of the confirmatory testing requirement and the modification of the pharmacogenetic report for CYP2C19, and provides analytical data demonstrating that the assay generates accurate results specific to the CYP2C19 variants reported in the results. Additionally, it was shown through robust user comprehension testing, previously reviewed and authorized under DEN180028, that the user can adequately interpret the results for the three variants, as well as understand interpretive drug information as presented in the pharmacogenetic report for CYP2C19.

5.8. CONCLUSION

The modifications to the 23andMe Personal Genome Service (PGS) Pharmacogenetic Reports are substantially equivalent to the predicate device DEN180028. As presented, the modified pharmacogenetic report for CYP2C19, without the confirmatory testing requirement, is a safe and effective consumer product that can safely and effectively assist customers with certain CYP2C19 genotypes in understanding how their body may respond to certain medications and encourage informed conversations with their healthcare provider.

Regulation Number and Section

§ 862.3364 Pharmacogenetic assessment system.

(a)
Identification. A pharmacogenetic assessment system is a qualitative in vitro molecular diagnostic system intended to detect nucleic acid variants isolated from human specimens for the purpose of assessing the presence of genetic variants that impact the metabolism, exposure, response, risk of adverse events, dosing, or mechanisms of prescription or over-the-counter medications. The intended use of the device must not include an indication for use in supporting or sustaining human life, being of substantial importance in preventing impairment of human health, or presenting a potential, unreasonable risk of illness or injury.(b)
Classification. Class II (special controls). A pharmacogenetic assessment system must comply with the following special controls:(1) Design verification and validation must include:
(i) Data appropriate, as determined by FDA, to demonstrate the analytical accuracy and reliability of the device in intended use specimens, including precision, reproducibility, accuracy, limits of detection, and interferences. This information must include:
(A) Data demonstrating appropriate, as determined by FDA, reproducibility for each genotype using each claimed sample type. Reproducibility data must be evaluated using specimens collected and processed in a manner consistent with the device's instructions for use, or, as determined by FDA, an appropriate alternative sample panel.
(B) Analytical data demonstrating the limits of detection, including the minimum amount of input deoxy-ribonucleic acid (DNA) that will consistently produce accurate results.
(C) Data demonstrating no clinically significant effects from endogenous and exogenous interferents relevant to each intended use specimen type. Interference data must also include an assessment of potentially interfering genetic sequences (
e.g., variants proximal to the variant of interest, pseudogenes).(D) Validation data appropriate, as determined by FDA, to support specimen collection and handling claims.
(E) Clinical data generated in intended use patient populations demonstrating the pharmacogenetic association between the genetic variant tested and any clinical claims or therapy-related recommendations associated with that genotype.
(ii) Results from an appropriate, as determined by FDA, user comprehension study that demonstrate the intended user can use the device safely. The user comprehension study must be designed to include the following:
(A) Study participants from a statistically sufficient sample size and a demographically diverse (
e.g., age, education level) population that is representative of the intended use population and naive to use of the device, and(B) An evaluation of all result comprehension concepts that are critical for safe use of the device.
(2) The labeling required under § 809.10 of this chapter must include:
(i) Clear information, written in language appropriate for the intended user, that describes instructions for how test results should be interpreted. These instructions must be supported by valid scientific evidence and include:
(A) Appropriate explanation of the claimed pharmacogenetic associations for all variants included in the test, any relevant variants not included in the test (
e.g., that may contribute to false negative results), and specific considerations by ethnicity, and(B) Appropriate explanation of non-genetic and non-tested genetic factors that may impact interpretation of the test results;
(ii) Detailed descriptions of analytical performance including, as applicable, precision, reproducibility, accuracy, limits of detection, and interferences as specified in paragraph (b)(1)(i) of this section, in language appropriate for the intended user;
(iii) A warning statement that the patient should not use the test results to stop or change any medication, and that medications should always be taken as prescribed by a healthcare professional;
(iv) A limiting statement explaining that this test is not intended to inform the patient about their current state of health, including whether the patient should or should not take a medication, or how much of a medication the patient should take, as appropriate;
(v) A warning statement that the test does not diagnose any health conditions and is not a substitute for visits to a doctor or other healthcare professional; and
(vi) A prominent and conspicuous limiting statement that the test provides only a preliminary test result that needs to be confirmed using an independent pharmacogenetic test without such a limitation prior to making any medical decisions. Alternatively, appropriate design verification and validation activities, including the generation of robust analytical data demonstrating appropriate analytical accuracy and reliability of test results for each genetic variant included in the test report, must be performed that demonstrate that the test can be used to make well-informed clinical decisions.
(3) The test report must include an appropriate description of how the test results should be used by healthcare providers who may receive the test results from their patients, as applicable.
(4) Publicly available pre-purchase labeling with unrestricted access that contains the following information must be provided:
(i) A clear description of the test and its technology, the genotypes detected, and relevant clinical claims associated with each genotype;
(ii) A clear description of what information the test will provide. This includes variant information, the limitations associated with the test, and any precautionary information about the test the user should be aware of before purchase; and
(iii) A discussion of answers to frequently asked questions that is sufficient to provide intended users with an appropriate understanding of information specific to each pharmacogenetic association that is claimed.
(5) The genetic test must use a sample collection device that is FDA-cleared or -approved, or classified as 510(k) exempt, with an indication for in vitro diagnostic use in DNA testing.
(6) The intended use of the device must not include an indication for use in supporting or sustaining human life, being of substantial importance in preventing impairment of human health, or presenting a potential, unreasonable risk of illness or injury.