23andMe® Personal Genome Service® (PGS®) Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants)

K223597 · 23AndMe, Inc. · QAZ · Aug 31, 2023 · Medical Genetics

Device Facts

• Record ID: K223597
• Device Name: 23andMe® Personal Genome Service® (PGS®) Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants)
• Applicant: 23AndMe, Inc.
• Product Code: QAZ · Medical Genetics
• Decision Date: Aug 31, 2023
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.6090
• Device Class: Class 2
• Attributes: Software as a Medical Device, PCCP

Intended Use

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 with the Oragene Dx model OGD500.001 for the purpose of reporting and interpreting genetic health risks, including the 23andMe PGS Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants).

Device Story

Device uses qualitative genotyping to detect 44 specific BRCA1/BRCA2 variants in human saliva. Saliva collected via Oragene-Dx device; DNA isolated and processed using multiplex microarray technology (Illumina BeadChip v5). DNA fragmented, hybridized to SNP-specific primers, extended with hapten-labeled nucleotides, and detected via fluorescently labeled antibodies. Tecan Evo and Illumina iScan systems perform extraction, processing, and scanning. Raw data analyzed by GenomeStudio and 23andMe proprietary Coregen software to determine genotypes. Results integrated into personalized reports for OTC user access. Reports provide information on detected variants and associated cancer risks (breast, ovarian, prostate, pancreatic). Device includes a Pre-Determined Change Control Plan (PCCP) for adding future validated variants. Output informs user-healthcare provider discussions; does not diagnose disease or determine treatment. Benefits include providing genetic risk information to individuals for proactive health management.

Clinical Evidence

Bench testing only. Method comparison study (n=samples from database) vs. bidirectional Sanger sequencing demonstrated >99% PPA and NPA. Precision study (intra-assay, inter-lot, inter-instrument, inter-operator, inter-day, inter-lab) showed 100% correct genotype calls. Minimum DNA input study validated performance across 5-50 ng/μL range. Pathogenicity of variants supported by peer-reviewed literature and ENIGMA consortium criteria.

Technological Characteristics

Qualitative genetic test for SNP detection. Sample: genomic DNA from saliva. Method: bead array hybridization, primer extension, fluorescent antibody detection. Instrumentation: Illumina iScan, GenomeStudio. Software: Coregen. PCCP implemented for variant expansion. Materials: Oragene Dx OGD500.001 collection kit.

Indications for Use

Indicated for reporting 44 specific BRCA1 and BRCA2 variants in adults ≥18 years. Provides genetic information regarding increased risk for breast, ovarian, prostate, and pancreatic cancers. Not for diagnostic use, treatment determination, or overall cancer risk assessment. Over-the-counter use.

Regulatory Classification

Identification

A qualitative in vitro molecular diagnostic system used for the detection of select variants in specified cancer-related genes. The device is intended to be used on genomic DNA isolated from human specimens collected by the user. The results of the test provide users with a genetic health risk assessment for developing certain cancers. The test may not include all variants associated with a predisposition of developing cancer and is not intended to describe a person’s overall risk of developing any type of cancer nor to aid in determination of treatment or act as a substitute for recommended cancer screenings or appropriate follow-up. The device is for over-the-counter use.

Special Controls

*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 <99 percent (incorrect test results compared to bidirectional sequencing or other methods identified as appropriate by FDA) must not be incorporated into test claims and reports. Accuracy measures generated from clinical specimens versus contrived samples or cell lines must be presented separately. Results must be summarized and presented in tabular format, by sample and by genotype.( *6* ) Point estimate of PPA for each genotype must be calculated as the number of correct calls for that genotype divided by the number of samples known to contain that genotype. The point estimate of NPA for each genotype must be calculated as the number of correct calls that do not contain that genotype divided by the number of samples known to not contain that genotype. 'No calls' must not be included in these calculations. Point estimates must be calculated along with 95 percent two-sided confidence intervals.(B) Precision and reproducibility data must be provided using multiple instruments and multiple operators, on multiple non-consecutive days, and using multiple reagent lots. The sample panel must include specimens from the claimed sample type ( *e.g.,* saliva) representing all genotypes for each variant (*e.g.,* wild type, heterozygous, and homozygous). Performance criteria must be predefined. A detailed study protocol must be created in advance of the study and then followed. The failed quality control rate must be indicated (*i.e.,* the total number of sample replicates for which a sequence variant cannot be called (no calls) or that fail sequencing quality control criteria divided by the total number of replicates tested). It must be clearly documented whether results were generated from clinical specimens, contrived samples, or cell lines. The study results must state, in a tabular format, the variants tested in the study and the number of replicates for each variant, and what conditions were tested (*e.g.,* number of runs, days, instruments, reagent lots, operators, specimens/type). The study must include all extraction steps from the claimed specimen type or matrix, unless a separate extraction study for the claimed sample type is performed. If the device is to be used at more than one laboratory, different laboratories must be included in the precision study (and reproducibility across sites must be evaluated). Any no calls or invalid calls in the study must be listed as a part of the precision and reproducibility study results.(C) Analytical specificity data: data must be provided evaluating the test performance ( *e.g.,* specimen extraction and variant detection) effect of potential endogenous and exogenous interferents relevant to the specimen type, and assessment of cross-contamination. Alternatively, for each suspected interfering mutation for which data is not provided demonstrating the effect of the interfering variant, the manufacturer must clearly identify the suspected interfering variants in the labeling to user test reports, and indicate that the impact the interfering variants may have on the test's performance has not been studied by providing a statement that reads, “It is possible that the presence of [insert identifying information for the suspected interfering variant] in a sample may interfere with the performance of this test. However, its effect on the performance of this test has not been studied.”(D) Analytical sensitivity data: data must be provided demonstrating the minimum amount of DNA that will enable the test to perform correctly in 95 percent of runs. (E) Device stability data: the manufacturer must establish upper and lower limits of input nucleic acid, sample, and reagent stability that will achieve the test's claimed accuracy and reproducibility. The manufacturer must evaluate stability using wild-type, heterozygous, and homozygous samples. Data supporting such claims must be provided. (F) Specimen type and matrix comparison data: specimen type and matrix comparison data must be generated if more than one specimen type can be tested with this device, including failure rates for the different specimens. (xiii) Clinical Performance Summary. (A) Information to support the clinical performance of each variant in the specific condition which is labeled as “are associated with increased risk” and reported by the test must be provided, as identified in paragraph (b)(4)(iii)(C) of this section. (B) Manufacturers must organize information by the specific variant combination as appropriate ( *e.g.,* wild type, heterozygous, homozygous, compound heterozygous, hemizygous genotypes). For each variant combination, information must be provided in the clinical performance section to support clinical performance for the risk category (*e.g.,* not at risk, increased risk). For each variant combination, a summary of key results must be provided in tabular format or using another method identified as appropriate by FDA to include the appropriate information regarding variant type, data source, definition of the target condition (*e.g.,* disease), clinical criteria for determining whether the target disease is present or absent, description of subjects with the target disease present and target disease absent (exclusion or inclusion criteria), and technical method for genotyping. When available, information on the effect of the variant on risk must be provided as the risk of a disease (lifetime risk or lifetime incidences) for an individual compared with the general population risk.(xiv) User comprehension study: information on a study that assesses comprehension of the test process and results by potential users of the test must be provided, including the following, as appropriate: (A) The test manufacturer must provide a genetic health risk education module to naïve user comprehension study participants prior to their participation in the user comprehension study. The module must define terms that are used in the test reports and explain the significance of genetic risk reports. (B) The test manufacturer must perform pre- and post-test user comprehension studies. The comprehension test questions must directly evaluate the material being presented to the user as described in paragraph (b)(3)(ii) of this section. (C) The manufacturer must provide a justification from a physician and/or genetic counselor that identifies the appropriate general and variant-specific concepts contained within the material being tested in the user comprehension study to ensure that all relevant concepts are incorporated in the study. (D) The user comprehension study must meet the following criteria: ( *1* ) The study participants must comprise a statistically sufficient sample size and demographically diverse population (determined using methods such as quota-based sampling) that is representative of the intended user population. Furthermore, the study participants must comprise a diverse range of age and educational levels and have no prior experience with the test or its manufacturer. These factors must be well-defined in the inclusion and exclusion criteria.( *2* ) All sources of bias (*e.g.,* non-responders) must be predefined and accounted for in the study results with regard to both responders and non-responders.( *3* ) The testing must follow a format where users have limited time to complete the studies (such as an on-site survey format and a one-time visit with a cap on the maximum amount of time that a participant has to complete the tests).( *4* ) Users must be randomly assigned to study arms. Test reports in the user comprehension study given to users must define the target condition being tested and related symptoms, explain the intended use and limitations (including warnings) for the test, explain the relevant ethnicities in regard to the variant tested, explain genetic health risks and relevance to the user's ethnicity, and assess participants' ability to understand the following comprehension concepts: the test's limitations, purpose, appropriate action, test results, and other factors that may have an impact on the test results.( *5* ) Study participants must be untrained, be naïve to the test subject of the study, and be provided the labeling prior to the start of the user comprehension study.( *6* ) The user comprehension study must meet the predefined primary endpoint criteria, including a minimum of a 90 percent or greater overall comprehension rate (*i.e.,* selection of the correct answer) for each comprehension concept. Other acceptance criteria may be acceptable depending on the concept being tested. Meeting or exceeding this overall comprehension rate demonstrates that the materials presented to the user are adequate for over-the-counter use.( *7* ) The analysis of the user comprehension results must include:( *i* ) Results regarding reports that are provided for each gene/variant/ethnicity tested;( *ii* ) Statistical methods used to analyze all data sets; and( *iii* ) Completion rate, non-responder rate, and reasons for nonresponse/data exclusion. A summary table of comprehension rates regarding comprehension concepts (*e.g.,* purpose of test, test results, test limitations, ethnicity relevance for the test results, appropriate actions following receipt of results) for each study report must be included.

Predicate Devices

• 23andMe PGS Risk Report for BRCA1/BRCA2 (Selected Variants) (DEN170046)

Submission Summary (Full Text)

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September 22, 2023

Image /page/0/Picture/1 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, which is a blue square with the letters "FDA" in white. To the right of the square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

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

Re: K223597

Trade/Device Name: 23andMe® Personal Genome Service® (PGS®) Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants) Regulation Number: 21 CFR 866.6090 Regulation Name: Cancer Predisposition Risk Assessment System Regulatory Class: Class II Product Code: OAZ

Dear Marianna Frendo:

The Food and Drug Administration (FDA) is sending this letter to notify you of an administrative change related to your previous substantial equivalence (SE) determination letter dated August 31, 2023. Specifically, FDA is updating this SE Letter to correct an error in the device trade name from 23andMe Personal Genome Service® (PGS®) Cancer Predisposition Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants) to 23andMe® Personal Genome Service® (PGS®) Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants) as an administrative correction.

Please note that the 510(k) submission was not re-reviewed. For questions regarding this letter please contact Shyam Kalavar, OHT7: Office of In Vitro Diagnostics, at 301-796-6807 or at Shyam.Kalavar(@fda.hhs.gov.

Sincerely.

# Shyam Kalavar -S

Shyam Kalavar Deputy Branch Chief Molecular Pathology and Cytology Branch Division of Molecular Genetics and Pathology OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

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Image /page/1/Picture/0 description: The image contains the logos of the Department of Health & Human Services and the Food and Drug Administration (FDA). The Department of Health & Human Services logo is on the left, and the FDA logo is on the right. The FDA logo is a blue square with the letters "FDA" in white, followed by the words "U.S. Food & Drug Administration" in blue.

August 31, 2023

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

Re: K223597

Trade/Device Name: 23andMe® Personal Genome Service® (PGS®) Cancer Predisposition Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants) Regulation Number: 21 CFR 866.6090 Regulation Name: Cancer Predisposition Risk Assessment System Regulatory Class: Class II Product Code: OAZ Dated: November 29, 2022 Received: December 2, 2022

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.

FDA's substantial equivalence determination also included the review and clearance of your Predetermined Change Control Plan (PCCP). Under section 515C(b)(1) of the Act, a new premarket notification is not

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required for a change to a device cleared under section 510(k) of the Act, if such change is consistent with an established PCCP granted pursuant to section 515C(b)(2) of the Act. Under 21 CFR 807.81(a)(3), a new premarket notification is required if there is a major change or modification in the intended use of a device, or if there is a change or modification in a device that could significantly affect the safety or effectiveness of the device, e.g., a significant change or modification in design, material, chemical composition, energy source, or manufacturing process. Accordingly, if deviations from the established PCCP result in a major change or modification in the intended use of the device, or result in a change or modification in the device that could significantly affect the safety or effectiveness of the a new premarket notification would be required consistent with section 515C(b)(1) of the Act and 21 CFR 807.81(a)(3). Failure to submit such a premarket submission would constitute adulteration and misbranding under sections 501(f)(1)(B) and 502(o) of the Act, respectively. Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

Your device is also subject to, among other requirements, the Quality System (OS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807): labeling (21 CFR Part 801 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 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 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-regulatory

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assistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

# Shyam Kalayar -S

Shyam Kalavar Deputy Branch Chief Molecular Pathology and Cytology Branch Division of Molecular Genetics and Pathology OHT7: Office of In Vitro Diagnostics 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) K223597

#### Device Name

23andMe Personal Genome Service (PGS) Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants)

#### Indications for Use (Describe)

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 with the Oragene Dx model OGD500.001 for the purpose of reporting and interpreting genetic health risks, including the 23andMe PGS Genetic Risk Report for BRCA1/BRCA2 (Selected Variants).

The 23andMe Personal Genome Service (PGS) Risk Report for BRCA1/BRCA2 (Selected Variants) is indicated for the reporting of the following 44 variants in the BRCA1 and BRCA2 genes.

BRCA1: c.68 69del, c.213-11T>G, c.427G>T, c.815 824dup, c.1556del, c.1687C>T, c.1961del, c.2681 2682del, c.2864C>A, c.3481 3491del, c.3598C>T, c.3627dup, c.3756 3759del, c.3770 3771del, c.4035del, c.4065 4068del.c.4327C>T.c.4357+1G>A.c.4964 4982del.c.4986+6T>G.c.5123C>A.c.5177 5180del.c.5266dup

BRCA2: c.658 659del, c.771 775del, c.2808 2811del, c.2957 2958insG, c.3170 3174del, c.3264dup, c.3545 3546del, c.3847 3848del, c.4471 4474del, c.5542del, c.5576 5579del, c.5682C>G, c.5946del, c.6037A>T, c.6275 6276del, c.7024C>T, c.7480C>T, c.7934del, c.8904del

The report describes if a person's genetic result is associated with an increased risk of developing breast cancer and ovarian cancer and may be associated with an increased risk for prostate cancer, and potentially other cancers. The variants included in this report do not represent the majority of the BRCA1/BRCA2 variants in people of most ethnicities. 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 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. 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.

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

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#### Traditional 510(k) Summary

# This summary of Traditional 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 Traditional 510(k) number is: K223597

#### Submitter/Primary Contact

Marianna Frendo, Manager, Requlatory Affairs 23andMe. Inc. 349 Oyster Point Blvd. SSF, CA 94080 [phone] [e-mail] Establishment Registration Number: 3007699459 Owner Operator Number: 10029404

#### Alternative Contact

Kathy Hibbs, Chief Administrative Officer 23andMe, Inc. 349 Oyster Point Blvd. SSF, CA 94080 [phone] [e-mail]

#### Date Prepared

29 August 2023

#### 5.0 PURPOSE OF SUBMISSION

In this Traditional 510(k) submission 23andMe seeks the following:

- 1. Clearance for an additional 41 BRCA1 and BRCA2 variants to be added to the existing authorized BRCA1/BRCA2 (Selected Variants) report, (DEN170046);
- 2. Establish a Pre-Determined Change Control Plan (PCCP) for reporting additional BRCA1 and BRCA2 variants and associated cancer risk information to the report.

This 510(k) summary describes the submission content supporting pre-market review for the proposed 41 additional variants to be added to the existing BRCA1/BRCA2 (Selected Variants) report, (DEN170046). This submission included a predetermined change control plan (PCCP) that was reviewed and authorized by FDA for adding additional

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validated BRCA1 and BRCA2 variants and associated cancer risk information to the 23andMe PGS® Cancer Predisposition Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants) without additional pre-market review. The PCCP outlined the specific protocols and acceptance criteria that 23andMe intends to use to clinically and analytically validate eligible BRCA1/BRCA2 variants.

# 5.1. REGULATORY INFORMATION

# Table 1. Proposed New Device

| Type of Submission: | Traditional 510(k) |
|-------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Common/Usual Name: | BRCA1/BRCA2 (Selected Variants) |
| Trade/proprietary Name: | 23andMe Personal Genome Service (PGS) Genetic<br>Health Risk Report for BRCA1/BRCA2 (Selected Variants) |
| Regulation Description: | A Cancer Predisposition Risk Assessment System is a<br>qualitative in vitro molecular diagnostic system used for<br>determining predisposition for cancer where the result of<br>the test may lead to prophylactic screening, confirmatory<br>procedures, or treatments that may incur morbidity or<br>mortality to the patient. The test could help to inform<br>conversations with a healthcare professional. This<br>assessment system is for over-the-counter use. This<br>device does not determine the person's overall risk of<br>developing any types of cancer. This test is not a<br>substitute for visits to a healthcare provider for<br>recommended screenings or appropriate follow-up and<br>should not be used to determine any treatments. |
| Regulation Number: | 21 CFR §866.6090 |
| Product Code: | QAZ |
| Class | Class II |
| Predicate Device: | 23andMe PGS Risk Report for BRCA1/BRCA2 (Selected<br>Variants), DEN170046, authorized on March 6, 2018 |

# 5.2. INTENDED USE

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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 with the Oragene Dx model OGD500.001 for the purpose of reporting and interpreting genetic health risks, including the 23andMe PGS Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants).

## 5.3. INDICATIONS FOR USE

The 23andMe Personal Genome Service (PGS) Risk Report for BRCA1/BRCA2 (Selected Variants) is indicated for the reporting of the following 44 variants in the BRCA1 and BRCA2 genes.

| Gene | Variant(s) |
|-------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| BRCA1 | c.68_69del, c.213-11T>G, c.427G>T, c.815_824dup, c.1556del, c.1687C>T, c.1960A>T, c.1961del, c.2681_2682del, c.2864C>A, c.3481_3491del, c.3598C>T, c.3627dup, c.3756_3759del, c.3770_3771del, c.4035del, c.4065_4068del, c.4327C>T, c.4357+1G>A, c.4964_4982del, c.4986+6T>G, c.5123C>A, c.5177_5180del, c.5266dup |
| BRCA2 | c.658_659del, c.771_775del, c.1929del, c.2808_2811del, c.2957_2958insG, c.3170_3174del, c.3264dup, c.3545_3546del, c.3847_3848del, c.4471_4474del, c.5542del, c.5576_5579del, c.5682C>G, c.5946del, c.6037A>T, c.6275_6276del, c.7024C>T, c.7480C>T, c.7934del, c.8904del |

The report describes if a person's genetic result is associated with an increased risk of developing breast cancer and ovarian cancer and may be associated with an increased risk for prostate cancer, pancreatic cancer, and potentially other cancers. The variants included in this report do not represent the majority of the BRCA1/BRCA2 variants in people of most ethnicities. 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 report is for over-thecounter use by adults over the age of 18, and provides genetic information to inform discussions with a healthcare professional. 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.

# 5.4. SUBSTANTIALLY EQUIVALENT PREDICATE DEVICE

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The components of the PGS are unchanged from the De Novo authorization for the predicate device, the PGS Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants) (DEN170046). These components include the saliva collection kit, reagents, beadchip, instrumentation, software, test processes and procedures.

The purpose of this Traditional 510(k) submission is to modify the 23andMe Personal Genome Service (PGS) Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants) to include 41 additional BRCA1 and BRCA2 variants to the existing authorized BRCA1/BRCA2 (Selected Variants) report (DEN170046) and to establish a Pre-Determined Change Control Plan (PCCP) for adding additional BRCA1 and BRCA2 variants and associated cancer risk information to the report.

The 23andMe Personal Genome Service (PGS) Risk Report for BRCA1/BRCA2 (Selected Variants) is similar to the predicate, having similar indications for use, the same intended use, and the same technological characteristics as its predicate device, with the exception of the implementation of a Predetermined Change Control Plan (PCCP) that specifies the protocols and acceptance criteria for making modifications to the reportable BRCA1/BRCA2 variants in a controlled manner such that the device is as safe and effective as the predicate.

Specific test methods for clinical and analytical validation are specified in the PCCP to establish substantial equivalence relative to DEN170046, and include sample size determination, analysis methods, and acceptance criteria. The Sponsor will perform testing of the additional BRCA1/2 variants according to the specified protocols, and if the validation data meet the specified acceptance criteria, they may add those variants to the BRCA1/2 report without additional premarket review.

The PCCP is limited to the addition of single nucleotide variants and small insertions and deletions (≤20 bp) in the BRCA1 and BRCA2 genes. The plan describes the specific clinical validation criteria that must be met to demonstrate that the new BRCA1/2 variants are high-risk, highly penetrant BRCA1/2 variants (i.e., those that are demonstrated to be linked to hereditary breast and ovarian cancer (HBOC) syndrome). Specific analytical validation protocols and acceptance criteria are also detailed in the plan to ensure that the device maintains the following performance characteristics for each new BRCA1/2 variant:

- Accuracy point estimates of ≥99% positive percent agreement (PPA) and negative . percent agreement (NPA), established by comparing the results of the 23andMe to bidirectional Sanger sequencing
- . ≥99% correct genotype calls assigned at each of two laboratory sites
- . ≥95% of samples vielding the correct genotype call at the minimum DNA input tested

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Additionally, software verification and validation activities are detailed in the PCCP and all must be completed successfully to modify the report to add the new BRCA1/2 variants. The plan specifies change control for genotype calling definitions and labeling updates to ensure the device remains as safe and effective as the predicate device.

Customers who previously opted-in to receive their BRCA1/2 report will receive an email notification informing them that the report has been updated. Customers will have access to their updated report unless they exercise their option to opt-out.

The PGS Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants) report is intended for over-the-counter, direct-to consumer use without prescription or physician order. As with the predicate device, the proposed modifications to the labeling of the BRCA1/BRCA2 (Selected Variants) report conform to submission requirements, and special controls of the original authorization (DEN170046). Before viewing the proposed modified labeling, customers must view an updated version of the BRCA1/BRCA2 (Selected Variants) education module that reflects the additional 41 variants, and customers with positive results are strongly advised to share their results with their healthcare provider. As instructed in the predicate device authorization DEN170046, the same healthcare provider limitations are included in the package insert:

- . 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.
- This test is intended to provide users with their genetic information, which may inform health-related lifestyle decisions and conversations with their doctor or other healthcare professional.
- · Any diagnostic or treatment decisions must be based on confirmatory prescription testing and/or other information that you determine to be appropriate for your patient, such as additional clinical testing and other risk factors that may affect individual risk and health care.

The Package Insert has been revised to incorporate information pursuant to the special controls agreed upon during DEN170046 - in particular, the addition of clinical and analytical validation information in support of the proposed additional 41 BRCA1/BRCA2 variants introduced in the modified BRCA1/BRCA2 (Selected Variants) report. Additionally, the Package Insert has been revised to provide a link to consumers to find a genetic counselor in the area in which they live. These revisions are consistent with the product classification under 21 CFR §866.6090. The proposed modifications are based on the additional test system performance data, developed according to the well

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established study protocols and acceptance criteria for 23andMe PGS cancer predisposition risk reports and submitted in this Traditional 510(k).

This submission proposes the same intended use for the Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants), does not introduce new risks to safety and effectiveness, and is supported by performance data collected for this purpose. As such, the proposed modifications to the labeling of the 23andMe Personal Genome Service Genetic Health Risk Report for BRCA1/BRCA2 (Selected Variants) are substantially equivalent to the predicate device authorized under DEN170046.

## 5.5. DEVICE DESCRIPTION

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 under K141410, which consists of a sealable collection tube containing a stabilizing buffer solution. Once the sample is collected, it is shipped to one of two 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, reagents and instrumentation manufactured by Illumina.

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 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 and clinically 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.

Engineering drawings, schematics, etc. of the 23andMe Personal Genome Service (PGS) BRCA1/BRCA2 (Selected Variants) report are not applicable to this device.

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# 5.6. 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(s): BRCA1, BRCA2 Variants: BRCA1 gene variants: c.68 69del, c.213-11T>G, c.427G>T, c.815 824dup, c.1556del, c.1687C>T. c.1960A>T. c.1961del, c.2681 2682del, c.2864C>A. c.3481 3491del, c.3598C>T, c.3627dup, c.3756 3759del, c.3770 3771del, c.4035del, c.4065 4068del, c.4327C>T, c.4357+1G>A, c.4964 4982del, c.4986+6T>G, c.5123C>A, c.5177 5180del, c.5266dup

BRCA2 gene variants: c.658 659del, c.771 775del, c.1929del, c.2808 2811del, c.2957 2958insG, c.3170 3174del, c.3264dup, c.3545 3546del, c.3847 3848del, c.4471 4474del, c.5542del, c.5576 5579del, c.5682C>G, c.5946del, c.6037A>T, c.6275 6276del, c.7024C>T, c.7480C>T, c.7934del, c.8904del

Genotyping principle: The DNA is fragmented and captured on a bead array by hybridization to immobilized SNP-specific primers, followed by extension with haptenlabeled 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, analyzed, and then integrated into pre-defined report templates specific for each condition associated with each genotype.

# Table 2. Substantial Equivalence

| Predicate<br>BRCA1/BRCA2<br>(Selected Variants)<br>Report<br>DEN170046 | Proposed modified<br>BRCA1/BRCA2<br>(Selected Variants)<br>Report<br>K223597 | |
|------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------|
| General Device Characterisitic Similarities | | |
| Intended<br>Use | The 23andMe Personal<br>Genome Service (PGS) | Same |
| | Predicate<br>BRCA1/BRCA2<br>(Selected Variants)<br>Report<br>DEN170046 | Proposed modified<br>BRCA1/BRCA2<br>(Selected Variants)<br>Report<br>K223597 |
| | uses qualitative<br>genotyping to detect<br>select clinically relevant<br>variants in genomic DNA<br>isolated from human<br>saliva collected from<br>individuals ≥18 years with<br>the Oragene Dx model<br>OGD500.001 for the<br>purpose of reporting and<br>interpreting genetic<br>health risks, including the<br>23andMe PGS Genetic<br>Health Risk Report for<br>BRCA1/BRCA2 (Selected<br>Variants). | |
| Collection<br>Kit | Oragene·Dx® saliva<br>collection device (OGD-<br>500.001)<br>K141410 | Same |
| BeadChip | Illumina Global Screening<br>Array customized for the<br>PGS. The chip is<br>designed to detect<br>specific single nucleotide<br>polymorphisms (SNPs)<br>as well as other genetic<br>variants; all markers refer<br>to specific positions in the<br>National Center for<br>Biotechnology<br>Information (NCBI) | Same |
| Predicate<br>BRCA1/BRCA2<br>(Selected Variants)<br>Report<br>DEN170046 | Proposed modified<br>BRCA1/BRCA2<br>(Selected Variants)<br>Report<br>K223597 | |
| reference human<br>genome. | | |
| Beadpool | Customized for 23andMe | |
| Instruments | Tecan Evo<br>Illumina iScan | |
| Software | Genome Studio<br>Coregen | |
| Specimen<br>Type | Saliva | |
| Differences | | |
| Measurand | 3 variants | |
| Reagents | Illumina Infinium HTS<br>Assay Reagents | |
| | Same | |
| | Same | |
| | Same | |
| | Same | |
| | 44 variants | |
| | Illumina Infinium HTS<br>Extra Assay Reagents | |

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# 5.7. PERFORMANCE TESTING SUMMARY

#### 5.7.1 Method Comparison (Accuracy)

23andMe performed a method comparison study to assess the accuracy of the 23andMe Personal Genome Service (PGS) test for the additional 41 variants to be added to the existing BRCA1/BRCA2 (Selected Variants) report. All 41 variants were included in this study.

The purpose of the study was to show equivalent genotype assignment between the 23andMe PGS assay and bidirectional Sanger sequencing. Samples were randomly selected from the 23andMe customer database based on their putative genotype. Genotyping of these samples was performed at a CLIA certified contract laboratory. All chosen samples were then tested using bidirectional Sanger sequencing. Genotyping results were compared between the 23andMe PGS assay and sequencing to calculate

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positive percent agreement (PPA) and negative percent agreement (NPA), with the sequencing results considered to be "truth". The passing criteria were greater than 99% PPA and greater than 99% NPA for each SNP.

This method comparison study vielded greater than 99% agreement. Therefore, the study passed the acceptance criteria of greater than 99% PPA and greater than 99% NPA. The method comparison study showed that the 23andMe assay is comparable to bidirectional Sanger sequencing.

# 5.7.2 Precision (Reproducibility)

23andMe performed a precision study to assess the repeatability and reproducibility of the 23andMe Personal Genome Service (PGS) test for the additional 41 variants to be added to the existing BRCA1/BRCA2 (Selected Variants) report. All 41 variants were included in this study.

This study evaluated intra-assay, inter-lot, inter-instrument, inter-operator, inter-day, and inter-lab precision. Samples were identified from the 23andMe customer database based on their putative genotype and 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, over 3 days, at each of 2 laboratory sites. Genotype results were confirmed using bidirectional Sanger sequencing. The passing criteria were a minimum of 99% correct genotype calls at each of two laboratory sites.

This precision study yielded 100% correct genotype calls for all samples across multiple days, operator teams, instruments, and reagent lots at 2 independent laboratory sites. Therefore, the study passed the acceptance criteria of at least 99% correct calls. In addition, the study had greater than 99% reproducibility and greater than 99% repeatability.

# 5.7.3 Minimum DNA Input (MDI)

23andMe performed a minimum DNA input study to assess the sensitivity of the 23andMe Personal Genome Service (PGS) test for the additional 41 variants to be added to the existing BRCA1/BRCA2 (Selected Variants) report. All 41 variants were included in this study.

Samples were identified from the 23andMe customer database based on their putative genotype. Each sample was diluted to 3 different concentrations and genotyped by the assay in a blinded fashion using 3 lots of reagents. Genotype results were confirmed using bidirectional Sanger sequencing. The minimum DNA requirement was defined as the lowest concentration at which at least 95% of samples yield the correct call.

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This minimum DNA input study yielded 100% correct genotype calls for all samples and 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. The performance requirement, specified by contract laboratory SOPs, is conservatively set at a minimum of 15 ng/μL and a maximum of 50 ng/μL. This minimum DNA input study demonstrated that the 23andMe assay is valid for samples with a DNA concentration range of 5 ng/μL to 50 nq/μL.

## 5.7.4 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.

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## 5.8. CLINICAL PERFORMANCE

The variants included in the modified BRCA1/BRCA2 (Selected Variants) report account for more than 90% of cancer-related BRCA1 and BRCA2 variants among people of Ashkenazi Jewish descent; about 30-40% among African Americans, people of European descent, and people of Hispanic or Latino descent; about 5-25% among people of East Asian Descent; and up to 35% among people of South Asian descent1. The pathogenicity of each variant is supported by multiple peer-reviewed studies and, in many cases, by the classification determination of an expert panel (ENIGMA²). In addition, most pathogenic BRCA1/2 variants are truncating; for those that are not, we rely on strong functional evidence demonstrating an impact on protein function.

# Table 3. Allele frequencies from 23andMe database and Genome Aggregation Database (gnomAD)

| Variant # | Gene | Variant name | rsID | Population | Allele frequencies<br>from 23andMe<br>(%)* | Allele<br>frequencies<br>from gnomAD<br>(%)** |
|--------------------|---------|---------------------|--------------|--------------------|--------------------------------------------|-----------------------------------------------|
| 1 | BRCA1 | c.68_69del | rs386833395 | European | 0.012% | 0.009% |
| | | | | African American | 0.004% | 0.000% |
| | | | | Ashkenazi Jewish | 0.445% | 0.405% |
| | | | | East Asian | 0.000% | 0.000% |
| | | | | Hispanic or Latino | 0.017% | 0.003% |
| | | | | South Asian | 0.025% | 0.013% |
| | | | | Middle Eastern | 0.007% | not available |
| 2 | BRCA1 | c.213-11T>G | rs80358061 | European | 0.003% | 0.003% |
| | | | | African American | <0.002% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| Variant # | Gene | Variant name | rsID | Population | Allele frequencies<br>from 23andMe<br>(%)* | Allele<br>frequencies<br>from gnomAD<br>(%)** |
| | | | | East Asian | 0.000% | 0.000% |
| | | | | Hispanic or Latino | 0.001% | 0.000% |
| | | | | South Asian | 0.000% | 0.000% |
| | | | | Middle Eastern | <0.007% | not available |
| 3 | BRCA1 | c.427G>T | rs80356991 | European | 0.001% | not observed in<br>gnomAD |
| | | | | African American | <0.002% | |
| | | | | Ashkenazi Jewish | 0.000% | |
| | | | | East Asian | 0.000% | |
| | | | | Hispanic or Latino | <0.001% | |
| | | | | South Asian | 0.000% | |
| | | | | Middle Eastern | 0.000% | |
| 4 | BRCA1 | c.815_824dup | rs387906563 | European | 0.000% | 0.000% |
| | | | | African American | 0.011% | 0.004% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | 0.000% | 0.000% |
| | | | | Hispanic or Latino | 0.003% | 0.003% |
| | | | | South Asian | 0.000% | 0.000% |
| | | | | Middle Eastern | 0.000% | not available |
| 5 | BRCA1 | c.1556del | rs80357662 | European | 0.001% | not observed in<br>gnomAD |
| | | | | African American | 0.000% | |
| Variant # | Gene | Variant name | rsID | Population | Allele frequencies<br>from 23andMe<br>(%)* | Allele<br>frequencies<br>from gnomAD<br>(%)** |
| | | | | Ashkenazi Jewish | 0.000% | |
| | | | | East Asian | 0.000% | |
| | | | | Hispanic or Latino | <0.001% | |
| | | | | South Asian | 0.000% | |
| | | | | Middle Eastern | 0.000% | |
| 6 | BRCA1 | c.1687C>T | rs80356898 | European | 0.002% | 0.006% |
| | | | | African American | 0.000% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | 0.000% | 0.000% |
| | | | | Hispanic or Latino | 0.001% | 0.000% |
| | | | | South Asian | 0.000% | 0.000% |
| | | | | Middle Eastern | 0.000% | not available |
| 7 | BRCA1 | c.1960A>T | rs80357355 | European | <0.001% | 0.006% |
| | | | | African American | 0.000% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | 0.000% | 0.000% |
| | | | | Hispanic or Latino | 0.001% | 0.000% |
| | | | | South Asian | 0.000% | 0.000% |
| | | | | Middle Eastern | 0.000% | not available |
| 8 | BRCA1 | c.1961del | rs80357522 | European | 0.001% | 0.002% |
| Variant # | Gene | Variant name | rsID | Population | Allele frequencies<br>from 23andMe<br>(%)* | Allele frequencies<br>from gnomAD<br>(%)** |
| | | | | African American | 0.000% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | 0.000% | 0.000% |
| | | | | Hispanic or Latino | <0.001% | 0.000% |
| | | | | South Asian | <0.006% | 0.000% |
| | | | | Middle Eastern | <0.007% | not available |
| 9 | BRCA1 | c.2681_2682del | rs80357971 | European | 0.002% | not observed in<br>gnomAD |
| | | | | African American | 0.000% | |
| | | | | Ashkenazi Jewish | 0.000% | |
| | | | | East Asian | 0.000% | |
| | | | | Hispanic or Latino | 0.001% | |
| | | | | South Asian | 0.000% | |
| | | | | Middle Eastern | 0.000% | |
| 10 | BRCA1 | c.2864C>A | rs80357295 | European | <0.001% | not observed in<br>gnomAD |
| | | | | African American | 0.000% | |
| | | | | Ashkenazi Jewish | 0.000% | |
| | | | | East Asian | 0.000% | |
| | | | | Hispanic or Latino | 0.001% | |
| | | | | South Asian | 0.000% | |
| | | | | Middle Eastern | 0.000% | |
| Variant # | Gene | Variant name | rsID | Population | Allele frequencies<br>from 23andMe<br>(%)* | Allele frequencies<br>from gnomAD<br>(%)** |
| 11 | BRCA1 | c.3481_3491del | rs80357877 | European | 0.001% | not observed in<br>gnomAD |
| | | | | African American | <0.002% | |
| | | | | Ashkenazi Jewish | 0.000% | |
| | | | | East Asian | 0.000% | |
| | | | | Hispanic or Latino | <0.001% | |
| | | | | South Asian | 0.000% | |
| | | | | Middle Eastern | 0.000% | |
| 12 | BRCA1 | c.3598C>T | rs62625307 | European | <0.001% | 0.000% |
| | | | | African American | 0.000% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | 0.000% | 0.000% |
| | | | | Hispanic or Latino | 0.001% | 0.000% |
| | | | | South Asian | <0.006% | 0.003% |
| | | | | Middle Eastern | 0.000% | not available |
| 13 | BRCA1 | c.3627dup | rs80357729 | European | 0.000% | 0.000% |
| | | | | African American | <0.002% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | 0.002% | 0.011% |
| | | | | Hispanic or Latino | 0.000% | 0.000% |
| | | | | South Asian | 0.000% | 0.000% |
| Variant # | Gene | Variant name | rsID | Population | Allele frequencies<br>from 23andMe<br>(%)* | Allele frequencies<br>from gnomAD<br>(%)** |
| 14 | BRCA1 | c.3756_3759del | rs80357868 | Middle Eastern | 0.000% | not available |
| | | | | European | 0.003% | 0.004% |
| | | | | African American | <0.002% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | 0.000% | 0.000% |
| | | | | Hispanic or Latino | 0.001% | 0.000% |
| | | | | South Asian | 0.000% | 0.000% |
| 15 | BRCA1 | c.3770_3771del | rs80357579 | Middle Eastern | <0.007% | not available |
| | | | | European | <0.001% | 0.001% |
| | | | | African American | <0.002% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | <0.002% | 0.000% |
| | | | | Hispanic or Latino | 0.000% | 0.000% |
| | | | | South Asian | <0.006% | 0.003% |
| 16 | BRCA1 | c.4035del | rs80357711 | European | 0.002% | 0.009% |
| | | | | African American | 0.000% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | 0.000% | 0.000% |
| | | | | Hispanic or Latino | <0.001% | 0.000% |
| Variant # | Gene | Variant name | rsID | Population | Allele frequencies<br>from 23andMe<br>(%)* | Allele<br>frequencies<br>from gnomAD<br>(%)** |
| | | | | South Asian | 0.000% | 0.000% |
| | | | | Middle Eastern | 0.000% | not available |
| 17 | BRCA1 | c.4065_4068del | rs80357508 | European | 0.002% | 0.002% |
| | | | | African American | <0.002% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | <0.002% | 0.000% |
| | | | | Hispanic or Latino | 0.002% | 0.000% |
| | | | | South Asian | 0.007% | 0.003% |
| | | | | Middle Eastern | <0.007% | not available |
| 18 | BRCA1 | c.4327C>T | rs41293455 | European | 0.002% | 0.004% |
| | | | | African American | <0.002% | 0.004% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | <0.002% | 0.000% |
| | | | | Hispanic or Latino | 0.002% | 0.008% |
| | | | | South Asian | 0.000% | 0.000% |
| | | | | Middle Eastern | <0.007% | not available |
| 19 | BRCA1 | c.4357+1G>A | rs80358027 | European | <0.001% | 0.000% |
| | | | | African American | 0.002% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | 0.000% | 0.000% |
| Variant # | Gene | Variant name | rsID | Population | Allele frequencies<br>from 23andMe<br>(%)* | Allele<br>frequencies<br>from gnomAD<br>(%)** |
| | | | | Hispanic or Latino | 0.000% | 0.000% |
| | | | | South Asian | 0.000% | 0.000% |
| | | | | Middle Eastern | <0.007% | not available |
| 20 | BRCA1 | c.4964_4982del | rs80359876 | European | 0.001% | 0.000% |
| | | | | African American | 0.000% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | | | East Asian | 0.000% | 0.000% |
| | | | | Hispanic or Latino | 0.001% | 0.000% |
| | | | | South Asian | 0.000% | 0.000% |
| | | | | Middle Eastern | 0.000% | not available |
| 21 | BRCA1 | c.4986+6T>G | rs80358086 | European | <0.001% | 0.001% |
| | | | | African American | 0.000% | 0.000% |
| | | | | Ashkenazi Jewish | 0.000% | 0.000% |
| | | |…