The 23andMe Personal Genome Service (PGS) uses qualitative genotyping to detect select clinically relevant variants in genomic DNA isolated from human saliva collected from individuals ≥18 years for the purpose of reporting and interpreting genetic health risks, including the 23andMe PGS Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13-Related). The 23andMe PGS Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13- Related) is indicated for reporting of the G84E variant in the HOXB13 gene. The report describes if a person has the G84E variant and if a male is at increased risk for prostate cancer. The variant included in this report is most common in people of European descent. The test report does not describe a person's overall risk of developing any type of cancer, and the absence of a variant tested does not rule out the presence of other variants that may be cancer-related. This test is not a substitute for visits to a healthcare provider for recommended screenings or appropriate follow-up and should not be used for diagnosis, to determine any treatments or medical interventions.

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 23andMe Personal Genome Service (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. It is a home use, over-thecounter (direct-to-consumer) DNA testing service intended to provide information and tools for consumers to learn about and explore their DNA.

Customer saliva is self-collected using the Oragene-Dx® Device manufactured by DNA Genotek, Inc. (previously cleared for carrier screening indications under K141410, and the same collection kit used to generate performance data for DEN140044, DEN160026, DEN170046, K182784, DEN180028, and K193492, which consists of a sealable collection tube containing a stabilizing buffer solution. Once the sample is collected, it is shipped to one of our Clinical Laboratory Improvement Amendments (CLIA) certified laboratories for testing.

DNA is isolated from the saliva and tested in a multiplex assay using a customized genotyping beadchip, and off the shelf reagents and instrumentation manufactured by Illumina. The multiplex assay simultaneously tests for more than 500,000 variants, including those for the previously authorized indications, as well as for the indications proposed herein.

Raw data is generated using Illumina GenomeStudio software, and then sent to 23andMe. The data is then 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 genetic health risk variant(s) have been detected in their sample and provide information about the disease 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 risks associated with the presence of a particular variant. 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 modified components of the Personal Genome Service included in this 510(k) submission are new labeling to include (a) one new variant to be reported, and (b) the qualitative reporting of one's Genetic Health Risk for Hereditary Prostate Cancer (HOXB13-Related).

Engineering drawings, schematics, etc. of Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13-Related) are not applicable to this device.

AI/ML Overview

The provided document describes the acceptance criteria and study proving the device meets these criteria for the 23andMe PGS Genetic Risk Report for Hereditary Prostate Cancer (HOXB13-Related).

Here's the breakdown of the information requested:

1. Table of Acceptance Criteria and Reported Device Performance

Performance Metric	Acceptance Criteria	Reported Device Performance
Method Comparison (Accuracy)	≥99% PPA and NPA for each SNP	>99% PPA and NPA for all genotypes. Study passed the criteria.
Precision / Reproducibility	≥99% correct calls	100% correct genotype calls. 100% reproducibility and repeatability.
DNA Input (Lowest Concentration)	≥95% correct calls at 5 ng/µL	100% correct genotype calls at 5, 15, and 50 ng/µL. Study passed.
Interfering Substance (Specificity)	100% accuracy when following IFU	100% accuracy when following instructions for use.
Labeling Comprehension	≥90% overall comprehension	Average comprehension rate ranged from 90.7% to 96.1%. Study met criteria.

2. Sample Sizes Used for the Test Set and Data Provenance

Accuracy/Method Comparison Study:
- Sample Size: Not explicitly stated as a number, but "Saliva samples were selected from the 23andMe customer biobank, based on their predetermined genotype and minimum volume required for testing." This implies a varied sample size based on the availability of specific genotypes.
- Data Provenance: From the "23andMe customer biobank" and "approved contract laboratory sites." The origin of the customers is not specified beyond "23andMe customer" which is a US-based company, suggesting primarily US data. The study was retrospective, using pre-existing samples from the biobank.
Precision Study:
- Sample Size: "DNA samples were selected based on their confirmed genotypes, and were obtained from the 23andMe biobank." Not an explicit number.
- Data Provenance: From the "23andMe biobank." Implies primarily US data, retrospective.
DNA Input Study:
- Sample Size: "DNA samples were obtained from the 23andMe biobank based on their listed genotypes." Not an explicit number.
- Data Provenance: From the "23andMe biobank." Implies primarily US data, retrospective.
Interfering Substance Study (referenced from DEN140044):
- Sample Size: Over 35,000 sample replicates.
- Data Provenance: Not explicitly stated for this particular study, but given it's for a US regulatory submission by a US company, it's highly likely to be US data, retrospective.
Labeling Comprehension Study (referenced from DEN160026):
- Sample Size: Not explicitly stated.
- Data Provenance: Not explicitly stated, but also likely US data.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts

Ground Truth Method: For the analytical studies (Method Comparison, Precision, DNA Input), the ground truth for genotyping was established by bi-directional Sanger sequencing.
Number/Qualifications of Experts: The document does not specify the number or qualifications of experts involved in performing or interpreting the Sanger sequencing results to establish the "truth." It only states that sequencing was performed "by an approved supplier" and that the sequencing results were "considered to be 'truth.'"

4. Adjudication Method for the Test Set (e.g., 2+1, 3+1, none)

The document does not describe any human adjudication method for establishing the ground truth from Sanger sequencing. It implies that the sequencing results themselves were directly taken as ground truth without further expert consensus or adjudication.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

No MRMC or comparative effectiveness study involving human readers (e.g., radiologists) with or without AI assistance was performed or described. This device is a direct-to-consumer genetic test, not an imaging-based AI diagnostic tool.
The closest concept is the "Labeling Comprehension" study, which assesses how well consumers understand the report. It indicates that the report and educational materials were effective in communicating relevant concepts for safe use. This is a measure of user comprehension, not human reader improvement with AI assistance.

6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was done

Yes, the performance studies (Accuracy, Precision, DNA Input, Interfering Substance) represent a standalone evaluation of the genotyping assay, which is essentially the "algorithm" or technical process of the device. The accuracy and precision figures are "algorithm only" performance metrics, as they compare the device's genotype calls directly against Sanger sequencing as the ground truth.

7. The Type of Ground Truth Used (expert consensus, pathology, outcomes data, etc.)

For the analytical performance studies (Accuracy, Precision, DNA Input), the ground truth for specific genetic variants (genotype) was established by bi-directional Sanger sequencing.
For the clinical performance, the document refers to "published studies of variant frequencies in various populations and the results of analytical studies" and "allele frequencies in the 23andMe customer database." This relies on established scientific literature and aggregated anonymized real-world data rather than individual outcomes or pathology reports.

8. The Sample Size for the Training Set

The document primarily describes validation studies (test sets) for the analytical performance of the device. It does not provide information about a separate "training set" sample size for developing the genotyping assay or the underlying "Coregen software." The genotyping method described relies on physical beadchip arrays and established principles of DNA analysis, not on a machine learning model that would typically have a distinct training phase with a dedicated dataset.
The "Customer biobank" is used for selecting samples for the performance studies, which may implicitly reflect data used in the development or refinement of their overall genotyping process, but it's not explicitly defined as a separate 'training set' for an AI model.

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

As mentioned above, the document does not elaborate on a distinct "training set" with established ground truth in the context of an AI/ML model for this genetic test. The "Coregen software" analyzes raw data from the beadchip, and its accuracy is validated against Sanger sequencing. The development process of this proprietary software, and any data used to "train" it (if it involves statistical modeling beyond simple rule-based interpretation of genotyping signals), is not detailed in terms of ground truth establishment.

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 is the Department of Health & Human Services logo. To the right of that is the FDA logo, with the letters "FDA" in a blue square. To the right of the square are the words "U.S. FOOD & DRUG ADMINISTRATION" in blue.

January 06, 2022

23andMe, Inc. Marianna Frendo Manager, Regulatory Affairs 349 Oyster Point Blvd. South San Francisco, CA 94080

Re: K211499

Trade/Device Name: 23andMe PGS Genetic Risk Report for Hereditary Prostate Cancer (HOXB13-Related) Regulation Number: 21 CFR 866.6090 Regulation Name: Cancer Predisposition Risk Assessment System Regulatory Class: Class II Product Code: QAZ Dated: May 12, 2021 Received: May 14, 2021

Dear Marianna Frendo:

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 statutes and regulations administered by other Federal agencies. You must comply with all the Act's

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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 (OS) 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 mediation-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,

Soma Ghosh, Ph.D. Chief Molecular Pathology and Cytology Branch Division of Molecular Genetics and Pathology 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) K211499

Device Name

23andMe Personal Genome Service (PGS) Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13-Related)

Indications for Use (Describe)

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

| | Prescription Use (Part 21 CFR 801 Subpart D)

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

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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

SUBMITTER / PRIMARY COMPANY

Marianna Frendo, Manager Regulatory Affairs 23andMe Inc. 349 Oyster Point Blvd SSF, CA 94080 mfrendo@23andme.com Establishment Registration Number: 3007699459 Owner Operator Number: 10029404

ALTERNATIVE CONTACT

Nikki Arora, Sr. Manager, Regulatory Affairs 349 Oyster Point Blvd SSF, CA 94080 nikkia@23andme.com

5.1. REGULATORY INFORMATION

Type of Submission:	Traditional 510k
Common/Usual Name:	Hereditary Prostate Cancer (HOXB13-Related)
Trade/proprietary Name:	23andMe Personal Genome Service (PGS) Genetic Health RiskReport for Hereditary Prostate Cancer (HOXB13-Related)
Regulation Description:	A Cancer Predisposition Risk Assessment System is aqualitative in vitro molecular diagnostic system used fordetermining predisposition for cancer where the result of the testmay lead to prophylactic screening, confirmatory procedures, ortreatments that may incur morbidity or mortality to the patient.The test could help to inform conversations with a healthcareprofessional. This assessment system is for over-the-counteruse. This device does not determine the person's overall risk ofdeveloping any types of cancer. This test is not a substitute forvisits to a healthcare provider for recommended screenings or

Table 5.1 Proposed new device

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	appropriate follow-up and should not be used to determine anytreatments.
Regulation Number:	21 CFR §866.6090
Product Code:	QAZ
Class	Class II

5.2. LEGALLY MARKETED EXISTING DEVICE

Trade Name: 23andMe Personal Genome Service (PGS)
Report Type	Carrier StatusReports	Genetic HealthRisk Reports	Genetic HealthRisk Reports	PharmacogeneticReports
ClassificationName	AutosomalRecessiveCarrierScreening GeneMutationDetectionSystem	Genetic VariantDetection andHealth RiskAssessmentSystem	CancerPredispositionRiskAssessmentSystem	Direct-to-ConsumerAccessPharmacogeneticAssessmentSystem
ClassificationPanel	Immunology /ClinicalChemistry	Immunology/Hematology	MolecularGenetics/Pathology	MolecularGenetics/Clinical Chemistry
RegulationNumber	21 CFR§866.5940	21 CFR§866.5950	21 CFR§866.6090	21 CFR §862.3364
Device Class	Class II	Class II	Class II	Class II
510(k)	Exempt	Exempt	Required	Required
Product Code	PKB	PTA	QAZ	QDJ
DEN/510(k)number	DEN140044	DEN160026	DEN170046,K182784	DEN180028,K193492
DecisionDate(s)	19Feb2015	06Apr2017	06Mar2018,18Jan2019	31Oct2018,17Aug2020

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5.3. LEGALLY MARKETED PREDICATE DEVICE(S)

23andMe Personal Genome Service (PGS)
Predicate	#1	#2
Report Name	Genetic Health Risk Report forBRCA1/BRCA2 (SelectedVariants)	Genetic Health Risk Report forMUTYH-Associated Polyposis (MAP)
ClassificationName	Cancer Predisposition RiskAssessment System	Cancer Predisposition RiskAssessment System
ClassificationPanel	Molecular Genetics/Pathology	Molecular Genetics/Pathology
RegulationNumber	21 CFR §866.6090	21 CFR §866.6090
Device Class	Class II	Class II
Product Code	QAZ	QAZ
DEN/510(k)number	DEN170046	K182784
DecisionDate(s)	06Mar2018	18Jan2019

5.5. 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.

Customer saliva is self-collected using the Oragene-Dx® Device manufactured by DNA Genotek, Inc. (previously cleared for carrier screening indications under K141410, and the same collection

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kit used to generate performance data for DEN140044, DEN160026, DEN170046, K182784, DEN180028, and K193492, which consists of a sealable collection tube containing a stabilizing buffer solution. Once the sample is collected, it is shipped to one of our Clinical Laboratory Improvement Amendments (CLIA) certified laboratories for testing.

Engineering drawings, schematics, etc. of Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13-Related) are not applicable to this device.

5.6. INDICATIONS FOR USE

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for visits to a healthcare provider for recommended screenings or appropriate follow-up and should not be used for diagnosis, to determine any treatments or medical interventions.

5.7. COMPARISON TO SUBSTANTIALLY EQUIVALENT PREDICATE DEVICES

BRCA1/BRCA2(SelectedVariants)DEN170046	MUTYHAssociatedPolyposisK182784	HereditaryProstate Cancer(HOXB13-Related)	Similaritiesanddifferences
Indicationsfor Use	The 23andMePersonal GenomeService (PGS) usesqualitativegenotyping todetect selectclinically relevantvariants in genomicDNA isolated fromhuman salivacollectedfrom individuals ≥18years with theOragene Dx modelOGD500.001 forthe purpose ofreporting andinterpreting genetichealth risks,including the23andMe PGSGenetic Health RiskReport forBRCA1/BRCA2(SelectedVariants)	The 23andMePersonal GenomeService (PGS) usesqualitativegenotyping todetect selectclinically relevantvariants in genomicDNA isolated fromhuman salivacollectedfrom individuals ≥18years with theOragene Dx modelOGD500.001 forthe purpose ofreporting andinterpreting genetichealth risks,including the23andMe PGSGeneticHealth Risk Reportfor MUTYHAssociatedPolyposis.	The 23andMePersonal GenomeService (PGS) usesqualitativegenotyping todetect selectclinically relevantvariants in genomicDNA isolated fromhuman salivacollected fromindividuals ≥18years with theOragene Dx modelOGD500.001 forthe purpose ofreporting andinterpreting genetichealth risks,including the23andMe PGSGenetic HealthRisk Report forHereditary ProstateCancer (HOXB13-Related).	Similar.

5.7.1 Intended use / Indications for use

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BRCA1/BRCA2(Selected Variants) isindicated forreporting of the185delAG and5382insC variantsin the BRCA1 geneand the 6174delTvariant in theBRCA2 gene. Thereport describes if awoman is atincreased risk ofdeveloping breastand ovarian cancer,and if a man is atincreased risk ofdeveloping breastcancer or may be atincreased risk ofdeveloping prostatecancer. The threevariants included inthis report are mostcommon in peopleof AshkenaziJewish descent anddo not representthe majority ofBRCA1/BRCA2variants in thegeneral population.The test reportdoes not describe aperson's overall riskof developing anytype of cancer, andthe absence of avariant tested doesnot rule out thepresence ofother variants that	MUTYH AssociatedPolyposis isindicated forreporting of theY179C and theG396D variants inthe MUTYH gene.The reportdescribes if aperson is atincreased risk ofdevelopingcolorectal cancer.The two variantsincluded in thisreport are mostcommon and beststudied in people ofNorthern Europeandescent and maynot represent themajority of theMUTYH variants inpeople of otherethnicities. The testreport does notdescribe a person'soverall risk ofdeveloping any typeof cancer, and theabsence of avariant tested doesnot rule out thepresence of othervariants that maybe cancer-related.This test is not asubstitute for visitsto a healthcareprovider forrecommendedscreenings or	Hereditary ProstateCancer (HOXB13-Related) isindicated forreporting of theG84E variant in theHOXB13 gene. Thereport describes ifa person has theG84E variant and ifa male is atincreased risk forprostate cancer.The variantincluded in thisreport is mostcommon in peopleof Europeandescent. The testreport does notdescribe a person'soverall risk ofdeveloping anytype of cancer, andthe absence of avariant tested doesnot rule out thepresence of othervariants that maybe cancer-related.This test is not asubstitute for visitsto a healthcareprovider forrecommendedscreenings orappropriate follow-up and should notbe used fordiagnosis, todetermine anytreatments or
may be cancer-related.This test is not asubstitute for visitsto a healthcareprovider forrecommendedscreenings orappropriate follow-up and should notbe used todetermineany treatments.	appropriate follow-up and should notbe used todetermineany treatments.	medicalinterventions.

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5.7.2 Technological Characteristics

	BRCA1/BRCA2(SelectedVariants)DEN170046	MUTYHAssociatedPolyposisK182784	HereditaryProstate Cancer(HOXB13-Related)	Similaritiesanddifferences
Test Type	Qualitative genetictest for singlenucleotidepolymorphismdetection	Qualitative genetictest for singlenucleotidepolymorphismdetection	Qualitativegenetic test forsingle nucleotidepolymorphismdetection	Same
SampleType/Matrix	Genomic DNAobtained from ahuman salivasample	Genomic DNAobtained from ahuman salivasample	Genomic DNAobtained from ahuman salivasample	Same
Gene	BRCA1and BRCA2	MUTYH	HOXB13	Different
Variant	185delAG and5382insC variantsin the BRCA1gene, and the6174delT variant	Y179C and G396D	G84E	Different
	in the BRCA2gene
SNP	i4000377,i4000378, andi4000379	rs34612342 andrs36053993	rs138213197	Different
GenotypingPrinciple	DNA is fragmentedand captured on abeadchip array byhybridization toimmobilized SNP-specific primers,followed byextension withhapten-labelednucleotides.Primers hybridizeadjacent to theSNPs and areextended with asingle nucleotidecorresponding tothe SNP allele.The incorporatedhapten-modifiednucleotides aredetected by addingfluorescentlylabeled antibodiesin several steps toamplify thesignals. Dataanalysis isperformed usingscatter plots	DNA is fragmentedand captured on abeadchip array byhybridization toimmobilized SNP-specific primers,followed byextension withhapten-labelednucleotides.Primers hybridizeadjacent to theSNPs and areextended with asingle nucleotidecorresponding tothe SNP allele.The incorporatedhapten-modifiednucleotides aredetected by addingfluorescentlylabeled antibodiesin several steps toamplify thesignals. Dataanalysis isperformed usingscatter plots	DNA isfragmented andcaptured on abeadchip array byhybridization toimmobilized SNP-specific primers,followed byextension withhapten-labelednucleotides.Primers hybridizeadjacent to theSNPs and areextended with asingle nucleotidecorresponding tothe SNP allele.The incorporatedhapten-modifiednucleotides aredetected byaddingfluorescentlylabeled antibodiesin several steps toamplify thesignals. Dataanalysis isperformed usingscatter plots	Same
Assay Components and instruments
Collection kit	Oragene.Dx®	Oragene.Dx®	Oragene.Dx®	Same
	saliva collectiondevice (OGD-500.001)K141410	saliva collectiondevice (OGD-500.001)K141410	saliva collectiondevice (OGD-500.001)K141410
Reagents	Illumina InfiniumHTS assayReagents	Illumina InfiniumHTS assayReagents	Illumina InfiniumHTS assayReagents	Same
Beadchip	Illumina InfiniumArray customizedfor the PGS. Thebeadchip array isdesigned to detectspecific singlenucleotidepolymorphisms(SNPs) as well asother geneticvariants; allmarkers refer tospecific positionsin the NationalCenter forBiotechnologyInformation (NCBI)reference humangenome	Illumina InfiniumArray customizedfor the PGS. Thebeadchip array isdesigned to detectspecific singlenucleotidepolymorphisms(SNPs) as well asother geneticvariants; allmarkers refer tospecific positionsin the NationalCenter forBiotechnologyInformation (NCBI)reference humangenome	Illumina InfiniumArray customizedfor the PGS. Thebeadchip array isdesigned todetect specificsingle nucleotidepolymorphisms(SNPs) as well asother geneticvariants; allmarkers refer tospecific positionsin the NationalCenter forBiotechnologyInformation(NCBI) referencehuman genome	Same
Instruments	Tecan EvoIllumina iScan withGenomeStudioSoftware	Tecan EvoIllumina iScan withGenomeStudioSoftware	Tecan EvoIllumina iScanwithGenomeStudioSoftware	Same
Software	Coregen	Coregen	Coregen	Same
Performance
Accuracy /MethodComparison	>99% PPA andNPA for allgenotypes	>99% PPA andNPA for allgenotypes	>99% PPA andNPA for allgenotypes	Same
Precision	>99%reproducibility and>99% repeatability	>99%reproducibility and>99% repeatability	>99%reproducibilityand >99%repeatability	Same
Minimum DNAInput	5ng/uL	5ng/uL	5ng/uL	Same
InterferingSubstance	100% accuracywhen following theinstructions for use	100% accuracywhen following theinstructions for use	100% accuracywhen followingthe instructionsfor use	Same
PotentiallyInterferingMutations	Warnings andLimitations:The performanceof these tests maybe affected by thepresence of raremutations. Theimpact ofpotentiallyinterferingmutations has notbeen evaluated	Warnings andLimitations:The performanceof these tests maybe affected by thepresence of raremutations. Theimpact ofpotentiallyinterferingmutations has notbeen evaluated	Warnings andLimitations:The performanceof these testsmay be affectedby the presenceof rare mutations.The impact ofpotentiallyinterferingmutations has notbeen evaluated	Same
MatrixComparison	Human SalivaOnly	Human SalivaOnly	Human SalivaOnly	Same
Stability	Specimen: 8 yearsReagents: 6-12MonthsCollection Kit: 24months	Specimen: 8 yearsReagents: 6-12MonthsCollection Kit: 24months	Specimen: 8yearsReagents: 6-12MonthsCollection Kit: 24months	Same
LabelingComprehension	90% or greateroverallcomprehensionrate	90% or greateroverallcomprehensionrate	90% or greateroverallcomprehensionrate	Same

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5.8. PERFORMANCE TESTING SUMMARY

All performance testing is managed by our ISO 13485:2016 certified Quality Management System, and conforms to design control requirements specified in 21CFR §820.30.

Analytical and Comprehension studies were performed and documented as described in Special Controls published in reclassification order DEN170046.

Software Verification and Validation studies were performed and documented as described in Guidance for the Content of Premarket Submissions for Software Contained in Medical Devices issued on 11Mav2005. 23andMe employs a variety of administrative, technical, and physical safequards in order to efficiently mitigate cybersecurity risks. 23andMe has been certified for compliance with ISO/IEC 27001:2013 Information Security Management System, including control implementation guidance within ISO/IEC 27018:2019 as well as the requirements and control implementation quidance within ISO/IEC 27701:2019 as a data controller. 23andMe is also actively implementing the controls and policies as defined in the "HITRUST CSF" (Common Security Framework), which includes the FDA recommended NIST Cybersecurity Framework (CsF).

5.8.1 Method Comparison (Accuracy)

23andMe performed a Method Comparison study to assess the accuracy of the 23andMe assay. The goal of the study was to show equivalent genotype assignment between the 23andMe assay and Sanger bi-directional DNA sequencing. Saliva samples were selected from the 23andMe customer biobank, based on their predetermined genotype and minimum volume required for testing. All assay genotyping was performed at approved contract laboratory sites. All chosen samples were then sequenced using Sanger bi-directional sequencing by an approved supplier. Genotyping results were compared between genotyping assay and bi-directional sequencing to calculate positive percent agreement (PPA) and negative percent agreement (NPA), with the sequencing results considered to be "truth." The passing criteria were a minimum of 99% PPA and NPA for each SNP. This method comparison study yielded >99% PPA and NPA for all genotypes. Therefore, the study passed the acceptance criteria of at least 99% PPA and NPA.

5.8.2 Precision / Reproducibility

23andMe performed a precision study to determine the reproducibility of the 23andMe assay. The goal of the study was to evaluate the following precision parameters of the assay: intra-assay, inter-lot, inter-instrument, inter-operator, inter-day, and inter-lab differences. DNA samples were selected based on their confirmed genotypes, and were obtained from the 23andMe biobank. To confirm, each sample was sequenced by bi-directional Sanger sequencing. The same samples were genotyped by the assay in a blinded fashion, with 3 lots of reagents, by multiple operator teams per day, using 3 different serial numbers of each of 2 instruments (Tecan and iScan), over 3 days, at each of 2 laboratory sites. Assay genotypes were compared with sequenced genotypes to determine the rates of correct genotype calls.

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This 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 at each of the two laboratory sites. There was no variation between any study conditions or any replicates for a given sample. Therefore, the study demonstrated 100% reproducibility and 100% repeatability for a given sample. Therefore, the study had 100% reproducibility and 100% repeatability.

5.8.3 DNA Input

23andMe performed a DNA input study for the genotyping assay. The design of the study was consistent with our previous agreement recorded in Q160823-A001 meeting minutes dated August 9, 2016. The goal of the study was to determine the lowest concentration of DNA that is necessary for successful assignment of the correct genotype. The DNA requirement for the PGS is defined as the lowest DNA concentration at which at least 95% of samples yielded the correct call. DNA samples were obtained from the 23andMe biobank based on their listed genotypes. Each sample was diluted to 3 different DNA concentrations and genotyped in a blinded fashion using 3 lots of reagents. To confirm the genotype call, each sample was also sequenced using bi-directional Sanger sequencing. Genotype results were compared with sequenced genotypes to determine the rates of correct genotype calls at each DNA concentration.

This study vielded 100% correct genotype calls for all samples and all reagent lots tested at sample DNA concentrations of 5, 15, and 50 ng/uL. Therefore, the study passed the acceptance criteria of 95% correct calls at a sample DNA concentration of 5 ng/μL. This study demonstrates that the assay can correctly genotype samples with a DNA concentration range of 5 ng/ul_to 50 ng/µL. The performance requirement specified by contracted laboratory SOPs is conservatively set at a minimum DNA requirement of 15 ng/μL.

5.8.4 Interfering Substance (Specificity)

The PGS assay requires the use of an FDA-cleared collection device (K141410). The device used for the test is the exact same device used for all carrier screening reports (DEN140044) and all genetic health risk reports (DEN160026 & DEN170046). A test for potentially interfering mutations was performed in support of DEN140044, and no new interfering substances have been identified. The cleared device includes labeling in the form of a package insert instructing the user to not eat, drink, smoke or chew gum for 30 minutes prior to collecting their saliva, thus minimizing the availability of potentially interfering substances in the saliva sample. 23andMe performed a series of studies to assess the effects of endogenous substances, exogenous substances, microbial substances, and smoking on the assay in support of submission DEN140044. Over these four studies, more than 35,000 sample replicates were tested, with no discordant or No Call results across 99 SNPs, for an accuracy of 100% when following the instructions for use (i.e., nothing by mouth for at least 30 minutes prior to collecting saliva).

5.8.5 Potentially Interfering Mutations

23andMe has identified one(1) potentially interfering mutation near G84E. The Company was unable to identify commercially available samples for these potentially interfering mutations.

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Therefore, a statement has been added to the limitations section of the package insert stating that the impact of interfering mutations has not been evaluated.

5.8.6 Matrix Comparison:

Human saliva is the only suitable sample matrix Comparison studies are not applicable.

5.8.7 Shelf life:

The PGS requires the use of the same FDA-cleared collection device and reagents that have been previously reviewed and authorized in K141410 and DEN140044.

5.8.8 Clinical Performance:

Clinical performance is based on published studies of variant frequencies in various populations and the results of analytical studies reported in this submission, as well as allele frequencies in the 23andMe customer database for the genetic variants and associations included in this submission.

The following table summarizes the clinical performance data for the PGS Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13-Related).

Table 5.3 Clinical Performance

HOXB13 G84E Allele Frequencies (%) in the 23andMe databaseª and the gnomAD database8

Ancestry group	23andMe	gnomAD
European	0.18%	0.76%c0.24%d
African American	0.04%	0.04%
Ashkenazi Jewish	<0.01%	0.01%
East Asian	0.00%	0.00%
Hispanic/Latino	0.06%	0.003%
South Asian	0.00%	0.00%
Middle Eastern	<0.01%	n/a

ª Based on approximately 5,552,000 individuals with European ancestry, 303,000 individuals with African ancestry, 168,000 individuals with Ashkenazi Jewish ancestry, 235,000 individuals with East Asian ancestry, 957,000 individuals with Hispanic/Latino ancestry, 57,000 individuals with South Asian ancestry, and 60,000 individuals with Middle Eastern ancestry. Because of the privacy considerations surrounding the use of customer data (namely, the risk of exposing the identity of individuals in the database), the frequencies provided to a hundredth of a percent and truncated at a minimum frequency it the number of individuals with a variant is fewer than five.

b Data was extracted from the gnomAD database: https://gnomad.broadinstitute.org/ accessed 09May2020.

C European (Finnish)

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ª European (Non-Finnish)

5.8.9 Labeling Comprehension

The key report message concepts for the Hereditary Prostate Cancer (HOXB13-Related) test were reviewed and determined to be the same as those identified and previously tested in the Genetic Health Risk device label comprehension study (DEN160026), which was determined suitable for the predicate devices (DEN170046 and K182784). The average comprehension rate per comprehension concept ranged from 90.7% to 96.1%, therefore the comprehension study met the predefined acceptance criteria of 90% or greater overall comprehension.

The representative Genetic Health Risk content and supporting educational materials were effective in communicating relevant concepts to users unfamiliar with genetic testing sufficient for the safe use of the PGS Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13-Related).

5.9. DISCUSSION

The PGS Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13-Related) has a similar intended use as the predicate devices, is not technologically different than the predicate devices, and therefore presents no new issues of safety or effectiveness when compared to the previously authorized predicate devices (DEN170046, K182784). Specifically, this submission for the PGS Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13-Related) includes analytical and clinical data demonstrating that the assay generates accurate results which the user can appropriately interpret.

5.10. CONCLUSION

The PGS Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13-Related) is substantially equivalent to the predicate devices DEN170046 (23andMe PGS Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants), authorized March 6, 2018) and K182784 (23andMe PGS Genetic Health Risk Report for MUTYH Associated Polyposis cleared on January 18, 2019). As presented, the PGS Genetic Health Risk Report for Hereditary Prostate Cancer (HOXB13-Related) is a safe and effective consumer product that can safely and effectively assist and encourage women and men with positive results to engage in potentially life-saving conversations with their healthcare provider.

Regulation Number and Section

§ 866.6090 Cancer predisposition risk assessment system.

(a)
Identification. A cancer predisposition risk assessment system is a qualitative in vitro molecular diagnostic system used for determining predisposition for cancer where the result of the test may lead to prophylactic screening, confirmatory procedures, or treatments that may incur morbidity or mortality to the patient. The test could help to inform conversations with a healthcare professional. This assessment system is for over-the-counter use. This device does not determine the person's overall risk of developing any types of cancer. This test is not a substitute for visits to a healthcare provider for recommended screenings or appropriate follow-up and should not be used to determine any treatments.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The labeling required under § 809.10 of this chapter and any pre-purchase page and test report generated, unless otherwise specified, must include:
(i) An intended use that specifies in the indications for use the genetic variants detected by the test. The specific variants must be appropriately validated as described in paragraphs (b)(4)(xii) and (b)(4)(xiii) of this section.
(ii) A section addressed to users with the following information:
(A) A warning statement accurately disclosing the genetic coverage of the test in lay terms, including information on variants not queried by the test, and the proportion of pathogenic variants in the genes that the assay detects in a specific population as identified in paragraph (b)(1)(i) of this section. The warning statement must indicate that the test [does not/may not, as appropriate] detect all genetic variants related to the genetic disease, and that the absence of a variant tested does not rule out the presence of other genetic variants that may impact cancer risk. The warning statement must also include the relevant population for which the variants reported by the test are most relevant.
(B) A limiting statement explaining that some people may feel anxious about getting genetic test health results. This is normal. If the potential user feels very anxious, such user should speak to his or her doctor or other healthcare professional prior to collection of a sample for testing. This test is not a substitute for visits to a doctor or other healthcare professional. Users should consult with their doctor or other healthcare professional if they have any questions or concerns about the results of their test or their current state of health.
(C) A limiting statement that a user's ethnicity may affect whether the test is relevant for them and may also affect how their genetic health results are interpreted.
(D) A warning statement that the test is not a substitute for visits to a healthcare professional for recommended screenings, and should not be used to determine any treatments or medical interventions.
(E) A warning statement that the test does not diagnose cancer or any other health conditions and should not be used to make medical decisions. The warning statement must indicate that the results should be confirmed in a clinical setting before taking any medical action.
(F) A limiting statement explaining that other companies offering a genetic risk test may be detecting different genetic variants for the same disease, so the user may get different results using a test from a different company.
(G) If applicable, a limiting statement that states the test does not test for variants in other genes linked to hereditary cancer.
(H) A limiting statement explaining that this test does not account for non-genetic factors and that other factors such as environmental and lifestyle risk factors may affect the risk of developing a given disease.
(I) Information to a potential purchaser or actual test report recipient about how to obtain access to a board-certified clinical molecular geneticist or equivalent to assist in pre- and post-test counseling.
(J) A limiting statement explaining that this test is not intended to tell you anything about your current state of health, or be used to make medical decisions, including whether or not you should take a medication or how much of a medication you should take.
(K) A limiting statement explaining that the laboratory may not be able to process a sample, and a description of the next steps to be taken by the manufacturer and/or the customer, as applicable.
(iii) A section in the labeling required under § 809.10 of this chapter and any test report generated that is for healthcare professionals who may receive the test results from their patients with the following information:
(A) A limiting statement explaining that this test is not intended to diagnose a disease, determine medical treatment or other medical intervention, or tell the user anything about their current state of health.
(B) A limiting statement explaining that this test is intended to provide users with their genetic information to inform health-related lifestyle decisions and conversations with their doctor or other healthcare professional.
(C) A limiting statement explaining that any diagnostic or treatment decisions should be based on confirmatory prescription testing and/or other information that is determined to be appropriate for the patient (
e.g., additional clinical testing and other risk factors that may affect individual risk and health care).(2) The genetic test must use a sample collection device that is FDA-cleared, -approved, or -classified as 510(k) exempt, with an indication for in vitro diagnostic use in over-the-counter DNA testing.
(3) The device's labeling must include a hyperlink to the manufacturer's public website where the manufacturer must make the information identified in paragraph (b)(3) of this section publicly available. The manufacturer's home page, as well as the primary part of the manufacturer's website that discusses the device, must provide a hyperlink to the web page containing this information and must allow unrestricted viewing access. If the device can be purchased from the website or testing using the device can be ordered from the website, the same information must be found on the web page for ordering the device or provided in a publicly accessible hyperlink on the web page for ordering the device. Any changes to the device that could significantly affect safety or effectiveness would require new data or information in support of such changes, which must also be posted on the manufacturer's website. The information must include:
(i) An index of the material being provided to meet the requirements in paragraph (b)(3) of this section and its location.
(ii) Technical information about the device, as specified in paragraph (b)(4) of this section.
(iii) A section that highlights summary information that allows the user to understand how the test works and how to interpret the results of the test. This section must, at a minimum, be written in plain language understandable to a lay user and include:
(A) Consistent explanations of the risk of disease associated with all variants included in the test, variants not included in the test, and specific considerations by ethnicity. If there are different categories of risk, the manufacturer must provide literature references and/or data that support the different risk categories. If there will be multiple test reports and multiple variants, the risk categories must be defined similarly among them. For example, “increased risk” must be defined similarly between different test reports and different variant combinations.
(B) Clear context for the user to understand the context in which the cited clinical performance data support the risk reported. This includes any risks that are influenced by ethnicity, age, gender, environment, and lifestyle choices.
(C) Materials that explain the main concepts and terminology used in the test that include:
(
1 ) Definitions: scientific terms that are used in the test reports.(
2 ) Pre-purchase page: this page must contain information that informs the user about what information the test will provide. This includes variant information, the condition(s) or disease(s) associated with the variant(s), professional guideline recommendations for general genetic risk testing, the limitations associated with the test (e.g., test does not detect all variants related to the disease), relevance of race/ethnicity, and any precautionary information about the test the user should be aware of before purchase. When the test reports the risk of a life-threatening or irreversibly debilitating disease or condition for which there are few or no options to prevent, treat, or cure the disease, a user opt-in page must be provided. This opt-in page must be provided for each disease type that falls into this category and must provide specific information relevant to each test result. The opt-in page must include:(
i ) An option to accept or decline to receive this specific test result;(
ii ) Specification of the risk involved if the user is found to have the specific genetic test result;(
iii ) Summary of professional guidelines that recommend when genetic testing for the associated target condition is or is not recommended;(
iv ) A recommendation to speak with a healthcare professional, genetic counselor, or equivalent professional before getting the results of the test;(
v ) The implications of receiving a no variants detected result; and(
vi ) The statement that the test does not diagnose cancer or any other health conditions and should not be used to make medical decisions. Results should be confirmed in a clinical setting before taking any medical action. Users should consult with a healthcare professional before taking any medical action.(
3 ) Frequently asked questions (FAQ) page: This page must provide information that is specific for each variant/disease pair that is reported. Information provided in this section must be scientifically valid and supported by corresponding peer-reviewed publications. The FAQ page must explain the health condition/disease being tested, the purpose of the test, the information the test will and will not provide, the relevance of race and ethnicity to the test results, information about the population to which the variants in the test is most applicable, the meaning of the result(s), other risk factors that contribute to disease, appropriate follow-up procedures, how the results of the test may affect the user's family, including children, and links to resources that provide additional information.(4) The device labeling must include a technical information section containing the following information:
(i) Gene(s) and variant(s) the test detects using standardized nomenclature, Human Genome Organization (HUGO) nomenclature and coordinates, as well as Single Nucleotide Polymorphism Database (dbSNP) reference SNP numbers (rs#).
(ii) A statement indicating that more than 1,000 variants in the BRCA1 and BRCA2 genes are known to increase cancer risk, as applicable.
(iii) Scientifically established disease-risk association of each variant detected and reported by the test. This risk association information must include:
(A) Genotype-phenotype information for the reported variants.
(B) When available, a table of expected frequency in the general population and different ethnicities, and risks of developing the disease in relevant ethnic populations and the general population.
(C) Information such as peer-reviewed published literature and/or professional guidelines used to determine what types and levels of evidence will distinguish whether the selected variants are reported as “are associated with increased risk” versus “may be associated with increased risk” of developing other cancers. All selected variants must be appropriately validated as required under paragraph (b)(1)(i) of this section. For selected variants reported as “are associated with increased risk,” the clinical evidence must be demonstrated with sufficient information (
e.g., professional guidelines and consistent associations in peer-reviewed published literature). For the selected variants reported as “may be associated with increased risk,” the clinical evidence must be reported in professional guidelines, but peer-reviewed published literature may not be consistent.(D) A statement about the current professional guidelines for testing these specific gene(s) and variant(s) for the specified disease(s).
(
1 ) If professional guidelines are available, provide the recommendations in the professional guideline(s) for the gene, variant, and disease for when genetic testing should or should not be performed, and cautionary information that should be communicated when a particular gene and variant is detected.(
2 ) If professional guidelines are not available, provide a statement that the professional guidelines are not available for these specific gene(s) and variant(s).(iv) The specimen type (
e.g., saliva, whole blood).(v) Assay steps and technology used.
(vi) Specification of required ancillary reagents, instrumentation, and equipment.
(vii) Specification of the specimen collection, processing, storage, and preparation methods.
(viii) Specification of risk mitigation elements and description of all additional procedures, methods, and practices incorporated into the directions for use that mitigate risks associated with testing.
(ix) Information pertaining to the probability of test failure (
e.g., percentage of tests that failed quality control) based on data from clinical samples, a description of scenarios in which a test can fail (e.g., low sample volume, low DNA concentration), how users will be notified of a test failure, and the nature of follow-up actions on a failed test to be taken by the user and the manufacturer.(x) When available, information specifying the probability of a false negative and false positive analytical result and any additional considerations by ethnicity.
(xi) Specification of the criteria for test result interpretation and reporting, including any distinctions between risk categories (
i.e., increased risk and greatly increased risk; are associated and may be associated).(xii) Information that demonstrates the performance characteristics of the test including:
(A) Accuracy of study results for each claimed specimen type.
(
1 ) Accuracy of the test must be evaluated with fresh clinical specimens collected and processed in a manner consistent with the test's instructions for use. If this is impractical, fresh clinical samples may be substituted or supplemented with archived clinical samples. Archived samples must have been collected previously in accordance with the instructions for use, stored appropriately, and randomly selected. In some limited circumstances, use of contrived samples or human cell line samples may also be appropriate and used as an acceptable alternative. The contrived or human cell line samples must mimic clinical specimens as much as is feasible and provide an unbiased evaluation of the test's accuracy.(
2 ) Accuracy must be evaluated by comparison to bidirectional Sanger sequencing or other methods identified as appropriate by FDA. Performance criteria for both the comparator method and the test must be pre-defined and appropriate to the test's intended use. Detailed study protocols must be provided.(
3 ) Information provided must include the number and type of specimens, broken down by clinically relevant variants for each indicated report that were compared to bidirectional sequencing or other methods identified as appropriate by FDA. The accuracy as positive percent agreement (PPA) and negative percent agreement (NPA) must be measured, and accuracy point estimates must be >99 percent (both per reported variant and overall). Uncertainty of the point estimate must be within an acceptable range, as identified by FDA, and must be presented using the 95 percent confidence interval.(
4 ) Sufficient specimens must be tested per genotype and must include all genotypes that will be included in the tests and reports. The number of samples tested in the accuracy study for each variant reported must be based on the variant frequency.(
5 ) Any no calls (i.e., absence of a result) or invalid calls (e.g., failed quality control) in the study must be included in accuracy study results and reported separately. The percent of final 'no calls' or 'invalid calls' must be clinically acceptable. Variants that have a point estimate for PPA or NPA of