← Product Code [QAZ](/submissions/MG/subpart-g%E2%80%94tumor-associated-antigen-immunological-test-systems/QAZ) · K182784

# MUTYH-Associated Polyposis (MAP) (K182784)

_23AndMe, Inc. · QAZ · Jan 18, 2019 · Medical Genetics · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/MG/subpart-g%E2%80%94tumor-associated-antigen-immunological-test-systems/QAZ/K182784

## Device Facts

- **Applicant:** 23AndMe, Inc.
- **Product Code:** [QAZ](/submissions/MG/subpart-g%E2%80%94tumor-associated-antigen-immunological-test-systems/QAZ.md)
- **Decision Date:** Jan 18, 2019
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 866.6090
- **Device Class:** Class 2
- **Review Panel:** Medical Genetics
- **Attributes:** Software as a Medical Device

## Indications for 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 MUTYHAssociated Polyposis. The 23andMe PGS Genetic Health Risk Report for MUTYHAssociated Polyposis is indicated for reporting of the Y179C and the G396D variants in the MUTYH gene. The report describes if a person is at increased risk of developing colorectal cancer. The two variants included in this report are most common and best studied in people of Northern European descent and may not represent the majority of the MUTYH variants in people of other 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 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.

## Device Story

Direct-to-consumer qualitative in vitro diagnostic test; uses self-collected saliva in Oragene-Dx device. DNA isolated and analyzed via multiplex microarray (Illumina Infinium HumanOmniExpress-24 BeadChip). Proprietary Coregen software determines genotypes for Y179C and G396D MUTYH variants. Results provided via user report; includes risk information for MUTYH-associated polyposis. Used by consumers; requires no physician intervention for sample collection. Healthcare providers use output to guide clinical screening or follow-up. Benefits include identifying increased colorectal cancer risk; enabling informed discussions with healthcare professionals.

## Clinical Evidence

Bench testing only. Method comparison study (BeadChip vs. Sanger sequencing) demonstrated >99% PPA and NPA. Precision/reproducibility study showed 100% correct genotype calls across multiple sites, operators, instruments, and reagent lots. DNA input study confirmed performance at 5ng/μL. Clinical performance estimated via allele frequencies in 23andMe database and published literature.

## Technological Characteristics

Qualitative SNP detection via Illumina Infinium HTS Assay. Genomic DNA from saliva. Bead array hybridization, primer extension with hapten-labeled nucleotides, fluorescent antibody detection. Illumina iScan/GenomeStudio instrumentation. Software: Coregen. Oragene-Dx collection device.

## Regulatory 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 Personal Genome Service (PGS) Risk Report for BRCA1/BRCA2 (Selected Variants) ([DEN170046](/device/DEN170046.md))

## Submission Summary (Full Text)

> This content was OCRed from public FDA records by [Innolitics](https://innolitics.com). If you use, quote, summarize, crawl, or train on this content, cite Innolitics at https://innolitics.com.
>
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number:
K182784

B. Purpose for Submission:
New device

C. Measurand:
Two variants (Y179C and G396D) in MUTYH gene

D. Type of Test:
Qualitative genetic test for detection of two variants in MUTYH gene

E. Applicant:
23andMe, Inc.

F. Proprietary and Established Names:
23andMe Personal Genome Service (PGS) Risk Report for MUTYH-Associated Polyposis (MAP)

G. Regulatory Information:
1. Regulation section:
21 CFR 866.6090
2. Classification:
Class II
3. Product code:
QAZ
4. Panel:
Pathology

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H. Intended Use:

1. Indication(s) for 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 MUTYHAssociated Polyposis. The 23andMe PGS Genetic Health Risk Report for MUTYHAssociated Polyposis is indicated for reporting of the Y179C and the G396D variants in the MUTYH gene. The report describes if a person is at increased risk of developing colorectal cancer. The two variants included in this report are most common and best studied in people of Northern European descent and may not represent the majority of the MUTYH variants in people of other 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 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.

2. Special conditions for use statement(s):

a. For over-the-counter (OTC) use.
b. The test does not diagnose cancer or any other health condition and should not be used to make medical decisions. Results should be confirmed in a clinical setting before taking any medical action.
c. This test is not a substitute for visits to a healthcare provider for recommended screening or appropriate follow-up. It is recommended that users consult with a healthcare provider if there are any questions or concerns about the test results or their current state of health.
d. The 23andMe PGS Genetic Health Risk Report for MUTYH-Associated Polyposis (MAP) detects only two variants in MUTYH gene and does not detect all genetic variants in this gene associated with increased risk of developing colorectal cancer (CRC). There are more than 100 variants in the MUTYH gene known to be associated with increased risk of developing cancer. The absence of a variant tested does not rule out the presence of other genetic variants that may be disease-related.
e. The test is intended for users ≥ 18 years old.
f. The laboratory may not be able to process a user's sample. The probability that the laboratory cannot process a sample can be up to 6-33%.
g. Three potentially interfering mutations near Y179C, and four potentially interfering mutations near G396D that are within the binding region for the variant being tested have been identified and are noted below. Interference due to these mutations was not tested.

|  MYTUH variant | Potentially Interfering Mutation  |
| --- | --- |
|  Y179C | rs190500741, rs533899702, rs201678305  |
|  G396D | rs559963863, rs529008617, rs3219490, rs531232542  |

h. A user's race, ethnicity, age, and sex may affect how the genetic test results are interpreted.

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i. It is important for the user to discuss their personal or family history of cancer with a healthcare professional. If the user has a personal or family history of cancer, or think they may have symptoms of cancer; the user should consult with their healthcare provider about appropriate testing.

j. Subject to meeting the limitations contained in the special controls under regulation 21 CFR 866.6090.

4. Special instrument requirements:

Tecan Evo, Illumina iScan and GenomeStudio system (qualified by 23andMe)

I. Device Description:

The 23andMe Personal Genome Service (PGS) is an over-the-counter (direct-to-consumer), qualitative in vitro diagnostic DNA test that provides genotype information using a customer's DNA.

Customer saliva is self-collected using the Oragene-Dx® Device manufactured by DNA Genotek, Inc. (previously cleared for carrier screening indications under K141410, which consists of a sealable collection tube containing a stabilizing buffer solution. Sample are shipped to one of two 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 and qualified by 23andMe.

The raw data is generated using Illumina GenomeStudio software and analyzed using the 23andMe's proprietary Coregen software, where a genotype is determined for each tested SNP. The results are used to generate reports for the customer that provide information about the detected genotypes.

Customer reports for the MUTYH variants include variant(s) detected and associated risk of cancer as reported in literature and guidelines. If no variant was detected, that information is also provided. The reports are designed to present scientific concepts to users in an easy-to-understand format, as well as the limitations of the testing and test results.

J. Substantial Equivalence Information:

1. Predicate device name(s):

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

2. Predicate 510(k) number(s):

DEN170046

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3. Comparison with predicate:

|  Similarities  |   |   |
| --- | --- | --- |
|  Item | Device | Predicate  |
|  Technology | Customized Illumina BeadChip | Same platform  |
|  Specimen type | Saliva | Same  |
|  Collection Device | Oragene-Dx® saliva collection device (OGD-500.001) | Same  |
|  Measurand | DNA | Same  |
|  Instruments and software | Tecan Evo, Illumina iScan and Genome Studio Coregen | Same  |
|  Differences  |   |   |
| --- | --- | --- |
|  Item | Device | Predicate  |
|  Variants to be reported | Y179C and G396D in MUTYH gene | 185delAG and 5382insC in BRCA1 and 6174delT in BRCA 2 gene  |
|  Indication | Qualitative reporting of risk for MUTYH-Associated Polyposis (MAP) and Colon Cancer | Qualitative reporting of risk for Breast Cancer  |

# K. Standard/Guidance Document Referenced (if applicable):

Not applicable

# L. Test Principle:

The assay uses multiplex microarray technology for the simultaneous detection of variants in human DNA. The BeadChip v5 assay (Illumina Infinium HumanOmniExpress-24 format chip) consists of silicon wafers etched to form wells loaded with silica beads, on which oligonucleotide capture probes are immobilized. DNA from saliva is fragmented and captured on a bead array by hybridization to immobilized SNP-specific primers, followed by extension with hapten-labeled nucleotides. The primers hybridize adjacent to the SNPs and are extended with a single nucleotide corresponding to the variant allele. The incorporated hapten-modified nucleotides are detected by adding fluorescently labeled antibodies in several steps to amplify the signals. The Tecan Evo and Illumina iScan instruments are used for extraction and processing of the DNA, and the BeadChip for scanning and quantification of the results. The genotype content is separated, analyzed, and then integrated into predefined report templates specific for each condition associated with each genotype. Genotypes are determined using the GenomeStudio and Coregen software packages. For the 23andMe PGS Genetic Health Risk Report for MUTYH-Associated Polyposis (MAP), information on two specific variants (Y179C, c.536A&gt;G in exon 7 and G396D, c.1187G&gt;A in exon 13) in the MUTYH gene are integrated into the report.

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# M. Performance Characteristics (if/when applicable):

# 1. Analytical performance:

# a. Precision/Reproducibility:

Reproducibility studies were conducted for the two variants reported in MUTYH gene. The reproducibility studies were designed to determine the imprecision due to assay run, lot, instrument, operator, day and site. DNA samples were procured and genotyped in blinded fashion. Genotypes of the DNA samples were confirmed through bidirectional Sanger sequencing. The study included one unique specimen for each genotype of the marker (i.e., specimens representing wild-type, heterozygous and homozygous cases for each allele). Each sample was run in triplicates and genotyped at the two laboratory sites on three days using three laboratory operator teams at each site, three lots of reagents (chosen at random from all available), three Tecan instruments, and three iScan instruments. Total number of replicates per sample is 162 except Y179C. Due to the rarity of Y179C marker, the "CC" genotype was tested with 2 instruments (Tecan and iScan), over 3 days, at each of 2 laboratory sites  $(N = 36)$ .

Among samples with valid calls, the precision study yielded  $100\%$  correct genotype calls including samples that were rerun. Information regarding samples that failed quality control (FQC) was also evaluated (as listed in Table 1). The data presented below are based on FQCs following a single run. Samples with FQC on the first run are re-tested in accordance with the laboratory protocol (i.e., one repeat test following invalid result). The results are shown in Table 1

Tables 1A-B. Precision Study Results Stratified by Site and Genotype
Table 1A. MUTYH Y179C (i5012768) Results

|  Genotype | Number of Replicates (including FQCs) | Number of Correct Calls | Number of Incorrect Calls | Number of FQCs | Percentage of FQCs  |
| --- | --- | --- | --- | --- | --- |
|  Site 1  |   |   |   |   |   |
|  Homozygous Common | 81 | 81 | 0 | 0 | 0%  |
|  Heterozygous | 81 | 81 | 0 | 0 | 0%  |
|  Homozygous Rare | 18 | 18 | 0 | 0 | 0%  |
|  Total | 180 | 180 | 0 | 0 | 0%  |
|  Site 2  |   |   |   |   |   |
|  Homozygous Common | 81 | 80 | 0 | 1 | 0%  |
|  Heterozygous | 81 | 73 | 0 | 8 | 10%  |
|  Homozygous Rare | 18 | 12 | 0 | 6 | 33%  |
|  Total | 180 | 165 | 0 | 15 | 8%  |

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Table 1B. MUTYH G396D (rs36053993) Results

|  Genotype | Number of Replicates (including FQCs) | Number of Correct Calls | Number of Incorrect Calls | Number of FQCs | Percentage of FQCs  |
| --- | --- | --- | --- | --- | --- |
|  Site 1  |   |   |   |   |   |
|  Homozygous Common | 81 | 81 | 0 | 0 | 0%  |
|  Heterozygous | 81 | 81 | 0 | 0 | 0%  |
|  Homozygous Rare | 81 | 81 | 0 | 0 | 0%  |
|  Total | 243 | 243 | 0 | 0 | 0%  |
|  Site 2  |   |   |   |   |   |
|  Homozygous Common | 81 | 80 | 0 | 1 | 0%  |
|  Heterozygous | 81 | 75 | 0 | 6 | 7%  |
|  Homozygous Rare | 81 | 73 | 0 | 8 | 10%  |
|  Total | 243 | 228 | 0 | 15 | 6%  |

# b. Linearity/assay reportable range:

Not applicable.

# c. Traceability, Stability, Expected values (controls, calibrators, or methods):

The assay requires two types of controls: the sample processing control and the reproducibility control. The information provided demonstrates that the sample processing control is stable for up to three months and the reproducibility control is stable for up to 12 months. See DEN140044 for detailed information.

# d. Detection limit:

The Limit of Detection (LoD) study was performed to determine the lowest concentration of DNA that is necessary for successful assignment of the correct Y179C and G396D variants using the 23andMe PGS test. Study samples were obtained from an external vendor and the 23andMe biobank based on their listed genotypes and included both homozygous and heterozygous common genotypes for each variant. Each sample, including three replicates per sample, was diluted to three different DNA concentrations (5, 15, and  $50\mathrm{ng} / \mu \mathrm{l}$ ) and genotyped by the PGS test in a blinded fashion using 3 lots of reagents. To confirm the genotype call, each sample was sequenced by bidirectional Sanger sequencing. Genotype calls from the test were compared with genotypes from Sanger sequencing to determine the rates of correct genotype calls at each DNA concentration.

The LoD was defined as the lowest DNA concentration at which at least  $95\%$  of samples yielded the correct call. This study yielded  $100\%$  correct calls per genotype for all samples across all reagent lots, at all sample concentrations tested. Therefore, the study passed the acceptance criteria of  $95\%$  correct calls at the lowest

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concentration tested (5 ng/μL). The performance requirement for the PGS Test, specified in the laboratory SOPs, is set at a minimum of 15 ng/μL DNA and maximum of 50 ng/μL DNA.

e. Analytical specificity:

Interfering Substances – Endogenous and Exogenous Substances

A series of studies were conducted to assess the effects of endogenous substances, exogenous substances, microbial substances, and smoking on the 23andMe PGS Test. The results of the Endogenous and Exogenous Interference studies can be found in the Decision Summary for DEN140044. No interference was observed with the endogenous substances tested. The study indicated that saliva samples should be collected at least 30 minutes after eating, drinking, chewing gum, using mouthwash.

Interfering Mutations

Analyses were performed to identify potentially interfering variants within the 50-nucleotide probe-binding regions of the two MUTYH variants detected by the test. Three potentially interfering mutations near Y179C, and four potentially interfering mutations near G396D that are within the binding region for the variant being tested have been identified (see list in Table 2). The specific mutations potentially interfering with detection of each tested variant are noted below. Interference due to these mutations was not tested. Therefore it is listed in the limitation statements in the package insert.

Table 2 Potentially Interfering Mutations in MUTYH gene
|  MYTUH variant | Potentially Interfering Mutation  |
| --- | --- |
|  Y179C | rs190500741, rs533899702, rs201678305  |
|  G396D | rs559963863, rs529008617, rs3219490, rs531232542  |

Smoking and Microbial Interference

The effects of smoking before the saliva collection and microbial interference were performed. The studies indicated that saliva samples should be collected at least 30 minutes after smoking and there is no effect on the accuracy of the test by five microbes that maybe found in human saliva. See DEN140044 for additional information.

f. Assay cut-off:

Not applicable.

g. Specimen Stability

Saliva samples for testing are collected with the Oragene-Dx collection device. The claimed specimen stability is 12 months at ambient temperature. See K110701 for sample stability information.

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f. Shipping Stability:

Saliva samples are shipped for testing in the Oragene-Dx collection device. Environmental conditions experienced during shipping were simulated by subjecting samples to freeze-thaw cycles (samples stored at high temperatures that could be experienced during shipping were evaluated in specimen stability. The claimed shipping stability is up to three freeze-thaw cycles. See K110701 for sample shipping stability information.

2. Comparison studies:

a. Method comparison with Sanger Bidirectional Sequencing:

Accuracy was evaluated through calculation of agreement of the genetic variant determinations between the 23andMe PGS test MUTYH results and Sanger bidirectional sequencing (comparator) results. All Sanger bidirectional sequencing was performed at an independent laboratory site. Saliva samples were randomly selected from the 23andMe customer biobank based on predetermined genotypes and the minimum volume required for testing. All chosen samples were then genotyped using Sanger bidirectional sequencing. Genotyping results were compared between the PGS test and bidirectional sequencing to calculate percent agreements with the sequencing results used as the reference. The comparison study results for the two variants are shown in Table 3 below. The accuracy data generated for each test report met the Manufacturer's pre-defined acceptance criteria: a minimum of  $99\%$  positive percent agreement (PPA) and negative percent agreement (NPA) for each genotype.

Table 3. Percent Agreement for MUTYH Variants by Genotypes

|  Genotype by Sanger | PGS Test Genotype Call |   |   |   |   |   | %PPA | %NPA | 95% CI‡  |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |  Correct* | Incorrect* | No Call | FQC | FQC % | Total Sample #  |   |   |   |
|  MUTYH Y179C Homozygous Common | 25 | 0 | 0 | 0 | 0% | 25 | 100 | 100 | 86.3-100  |
|  MUTYH Y179C Heterozygous | 26 | 0 | 0 | 0 | 0% | 26 | 100 | 100 | 86.8-100  |
|  MUTYH Y179C Homozygous Rare | 14 | 0 | 0 | 1 | 7% | 15 | 100 | 100 | 76.8-100  |
|  MUTYH G396D Homozygous Common | 26 | 0 | 0 | 0 | 0% | 26 | 100 | 100 | 86.8-100  |
|  MUTYH | 27 | 0 | 0 | 0 | 0% | 27 | 100 | 100 | 87.2-100  |

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|  G396D Heterozygous |  |  |  |  |  |  |  |  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  MUTYH G396D Homozygous Rare | 25 | 0 | 0 | 0 | 0% | 25 | 100 | 100 | 86.3-100  |

*Relative to Sanger sequencing
†Clopper-Pearson exact method

b. Matrix comparison:

Not applicable. This test is for use with human saliva samples only.

3. Clinical studies:

a. Disease Description and Clinical Summary:

The MUTYH gene encodes for an enzyme called MYH glycosylase, which is involved in the repair of DNA. This enzyme corrects particular errors that are made when DNA is copied (DNA replication) in preparation for cell division. During normal cellular activities, guanine sometimes becomes altered by oxygen, which causes it to pair with adenine instead of cytosine. MYH glycosylase fixes this error so mutations do not accumulate in the DNA and lead to tumor formation. This type of repair is known as base excision repair and can lead to a type of cancer known as MUTYH-associated polyposis.

MUTYH-associated polyposis is considered to be autosomal recessive polyposis syndrome (one variant must be inherited by each parent), though monoallelic carriers may have slightly increased risk for colorectal cancer. Affected patients have multiple colorectal adenomas and an increased risk of colorectal cancer.

The two most common MUTYH gene pathogenic variants in Western Europeans and North Americans are Y179C (rs34612342, c.536A&gt;G in exon 7) and G396D (rs36053993, c.1187G&gt;A in exon 13) (Nielsen 2012; ClinVar). (previously referred to as Y165C and G382D, respectively). However, pathogenic variants at different loci have been reported in other populations. Patients with MUTYH-associated polyposis may be homozygous or compound heterozygous for these or other pathogenic variants in the MUTYH gene.

These two variants cover about 80-90% of all MUTYH pathogenic variants in northern European populations (GeneReviews, Nielsen 2012; Win 2014; Cleary 2009). The MUTYH carrier frequencies in this population are about 1-2%, from which a prevalence of 1:10,000 to 1:40,000 could be calculated (GeneReviews, Nielsen 2012). These two variants have also been observed in people of other ethnicities.

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Table 4 below listed the frequency of MUTYH variants in 23andme customers and public databases.

Table 4 Frequency of MUTYH variants in 23andMe customers

|  Variant Name | European | African American | Ashkenazi Jewish | East Asian | Hispanic or Latino | South Asian  |
| --- | --- | --- | --- | --- | --- | --- |
|  Y179C | 0.42% | 0.12% | 0.00% | 0.00% | 0.29% | 0.05%  |
|  G396D | 1.16% | 0.41% | 0.00% | 0.01% | 1.02% | 0.05%  |

Clinical validity of the variants is supported by published data and NCCN and ACG guidelines. Clinical data relating to pathogenic variants in MUTYH is summarized in Table 5 below.

Table 5 Health Risk Estimates and Test Interpretation for Colorectal Cancer

|  Genotype | Risk Estimates | References  |
| --- | --- | --- |
|  0 variant | 4.2% | SEER Cancer Statistics Review, 1975-20151  |
|  1 MUTYH variant | Uncertain to slightly increase risk | NCCN 20172; Win 20143  |
|  2 MUTYH variants | 43-100% | ACG Guidelines4, Nielsen 20125  |

Individuals with MUTYH-associated polyposis usually develop between 10 to 100 colorectal polyps by the fifth or sixth decade. Individuals with MUTYH-associated polyposis are at high risk for developing CRC, and approximately 60 percent of patients have CRC at presentation. In contrast, the risk of CRC in monoallelic MUTYH carriers appears to be only marginally increased with an estimated lifetime risk of slightly evaluated $^{2,3}$ .

# b. Other clinical supportive data:

# i. User Comprehension Study

User comprehension studies were performed to assess the comprehension of the Genetic Health Risk report. See DEN160026 supportive user comprehension studies.

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ii. Frequently Asked Questions Material

The Manufacturer has developed a Frequently Asked Questions (FAQ) section for the MUTYH-Associated Polyposis (MAP) Genetic Health Risk (GHR) report, which is included in the test report and accessible to the user on the Manufacturer’s public website. The FAQs are specific to the variants and disease risk associations being reported, where applicable. The FAQ section was created to provide users with information to adequately understand the purpose, limitations and meaning of the results of the test. The FAQ section was developed using methodology consistent with the Manufacturer’s labeling design, identification of primary communication messages, and label comprehension. The concepts covered in the FAQ section include: the test results, purpose of the test, limitations of the test, relevance of race and ethnicity on test results, meaning of the result, other risk factors that contribute to disease, appropriate follow-up procedures, how the results of the test may affect the user’s family and children, and links to resources that provide additional information. Additionally, the FAQ section provides definitions for terminology found in Genetic Health Risk Reports that is used to describe risks associated with detected variants.

iii. User Opt-In Page

Prior to receiving the test results, a pre-purchase page informs users that there is a choice of whether or not to receive the MUTYH-Associated Polyposis (MAP) test report. Users have an opportunity to opt into receiving these results after reviewing important information included in an opt-in page. The opt-in page is provided for the MUTYH-Associated Polyposis (MAP) GHR report users due to the nature of the diseases and associated risks for this report and the fact that this test is not designed to inform clinical decision-making. Users will be directed to a page entitled, “Choose your health reports” which provides the option to exclude this report from the users account. The report selection page includes important information to allow the users to make an informed decision. Results of the MUTYH-Associated Polyposis (MAP) report are locked by default, and will never be shown to users unless they have specifically chosen to receive the report at any time, including after results for other reports have been received.

4. Expected values/Reference range:

Not applicable.

N. Instrument Name:

Illumina iScan BeadChip scanner with GenomeStudio software (qualified by the laboratory)

O. System Descriptions:

1. Modes of Operation:

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Same as referenced in DEN140044

2. Software:

FDA has reviewed applicant’s Hazard Analysis and software development processes for this line of product types:

Yes ☐ X ☐ or No ☐

Level of Concern:

Moderate

Software Description:

Same as referenced in DEN140044

Revision Level History:

A software revision history record for the 23andMe software system software was acceptable.

Unresolved Anomalies:

There are no known unresolved anomalies associated with the system software.

EMC Testing:

Not applicable.

3. Specimen Identification:

Same as referenced in DEN140044

4. Specimen Sampling and Handling:

Same as referenced in DEN140044

5. Calibration:

Same as referenced in DEN140044

6. Quality Control:

Same as referenced in DEN140044

P. Other Supportive Instrument Performance Characteristics Data Not Covered In The

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"Performance Characteristics" Section above:

Refer to K141410 for saliva collection device details and study results.

Q. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Parts 801 and 809, as applicable, and the special controls for this device type.

R. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

---

**Source:** [https://fda.innolitics.com/submissions/MG/subpart-g%E2%80%94tumor-associated-antigen-immunological-test-systems/QAZ/K182784](https://fda.innolitics.com/submissions/MG/subpart-g%E2%80%94tumor-associated-antigen-immunological-test-systems/QAZ/K182784)

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