K Number

Device Name

23ANDME PERSONAL GENOME SERVICE (HEREINAFTER KNOWN AS PGS)

Manufacturer

23andMe

Date Cleared

2015-02-19

(266 days)

Product Code

PKB

Regulation Number

866.5940

Intended Use

The 23andMe PGS Carrier Screening Test for Bloom Syndrome is indicated for the detection of the BLMAsh variant in the BLM gene from saliva collected using an FDA cleared collection device (Oragene DX model OGD-500.001). This test can be used to determine carrier status for Bloom syndrome in adults of reproductive age, but cannot determine if a person has two copies of the BLM140 variant. The test is most relevant for people of Ashkenazi Jewish descent.

Device Description

The 23andMe Personal Genome Service (PGS) Carrier Screening Test for Bloom Syndrome (hereafter the "PGS") is a non-invasive genetic information service that combines qualitative genotyping data for an individual. The PGS is indicated for use for the detection of the BLM4sh variant in the BLM gene from saliva collected using the Oragene•Dx Saliva Collection Device (Oragene Dx model OGD-500.01). The core components of the PGS consist of the saliva collection kit: custom genotyping chip: laboratory procedures, equipment and analysis; and result reporting software.

More Information

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Type

Center

Panel

Date Received

Product Code

Subsequent Product Codes

Applicant Name (Manufacturer)

Device Name

Decision

Street 1

Street 2

City

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Country Code

ZIP

Postal Code

Class Advise Committee

SSP Indicator

Third Party Reviewed

Expedited Review

Predicate Devices

No predicate device exists.

Reference Devices

K141410

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a genetic test using genotyping chips and standard laboratory procedures for data analysis and reporting. There is no mention of AI or ML being used in the data processing or interpretation.

Therapeutic

No.
This device is a genetic carrier screening test to detect a specific gene variant. It is used to determine carrier status for Bloom syndrome and does not treat or prevent any medical condition.

Diagnostic

Yes

The device is indicated for the "detection of the BLMAsh variant in the BLM gene" and can "determine carrier status for Bloom syndrome in adults of reproductive age," which are diagnostic purposes.

SaMD (Software as a Medical Device)

The device description explicitly states that the core components include a saliva collection kit, custom genotyping chip, laboratory procedures, equipment, and analysis, in addition to the result reporting software. This indicates significant hardware and physical components are integral to the device's function.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The intended use explicitly states that the device is for the "detection of the BLMAsh variant in the BLM gene from saliva". This is a diagnostic purpose, specifically for determining carrier status for Bloom syndrome.
Sample Type: The test is performed on saliva, which is a biological specimen collected from the human body.
Purpose: The purpose is to provide information about an individual's genetic makeup related to a specific condition (Bloom syndrome carrier status). This information is used for health-related purposes (determining reproductive risk).
Device Description: The description details the components involved in the testing process, including the saliva collection kit, genotyping chip, laboratory procedures, and analysis. These are all elements of an in vitro diagnostic system.
Performance Studies: The document includes detailed performance studies (Precision, Reproducibility, Detection Limit, Interference, Method Comparison) which are standard requirements for demonstrating the analytical and clinical validity of an IVD.
Key Metrics: The document reports key metrics like Positive Percent Agreement (PPA), Negative Percent Agreement (NPA), and Overall Agreement, which are used to evaluate the performance of diagnostic tests.

While the device is intended for over-the-counter (OTC) use and the results are reported directly to the user, the underlying process of analyzing a biological sample to provide diagnostic information about a health condition firmly places it in the category of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

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

PKB

Device Description

The saliva collection kit includes a sample collection tube with a unique barcode printed by the manufacturer, funnel, preservative solution, instructions for use, and pre-paid packaging for returning the sample to the processing laboratory. Saliva may be collected by spitting directly into the Oragene Dx container or may be transferred into the Oragene Dx container using a sponge. Saliva samples collected using Oragene•Dx are stabilized and can be transported and/or stored long term at ambient conditions.

The PGS is indicated for the detection of the BLMAsh variant in the BLM gene using DNA extracted from 2 mL saliva samples that are collected in a FDA cleared or approved collection device. Illumina manufactures a custom Infinium BeadChip genotyping chip for the device. The chip is designed to detect specific single nucleotide polymorphisms (SNPs) as well as other genetic variants; all markers refer to specific positions in the National Center for Biotechnology Information (NCBI) reference human genome.

After placing an order, an individual receives via post an Oragene-Dx saliva collection kit. Once the saliva sample is received by the laboratory. DNA extraction and quantitation steps occur. Samples meeting a minimum DNA concentration of 15 ng/uL are processed and prepared for amplification and BeadChip addition. BeadChips are read by the Illumina iScan, which is a laser-based, high-resolution optical imaging system. The instrument reads BeadChips by employing red and green lasers to excite the fluorophores of the allele-specific extended products found on the beads. Light emissions from these fluorophores are then recorded in high-resolution images of each BeadChip section. Data from these are analyzed to determine genotypes using Illumina's GenomeStudio software package. GenomeStudio is a modular software application that allows viewing and analyzing of genotypic data obtained from the iScan.

Mentions image processing

Yes

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Adults of reproductive age.

Intended User / Care Setting

For over-the-counter (OTC) use.

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

Not Found

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

Precision Study with Human Cell Line Samples:

Sample size: Six DNA samples from human cell lines (4 BLM homozygous common ("DD"), 1 BLMAs" heterozygous ("DI"), and 1 BLMAsh homozygous rare ("II")).
Data source: Human cell lines. Each sample was also sequenced by bi-directional Sanger Sequencing to confirm the BeadChip genotype.
Annotation protocol: Samples were tested over 5 days by 3 operator teams at site 1 and 3 operator teams at site 2, using 4 lots of reagents and 3 Tecan and 3 iScan instruments in different combinations. Information regarding samples that failed quality control (FQC) was evaluated.

Laboratory Reproducibility Study with Saliva Samples:

Sample size: 105 BLMAsh homozygous common ("DD") saliva samples obtained from individuals using the 23andMe Saliva Collection kit (Oragene-DX, OGD500.001, saliva collection kit).
Data source: Human saliva samples.
Annotation protocol: Sample processing was performed at two sites and tested with the PGS test for Bloom syndrome. Fifty samples were initially processed at the first site and 55 samples at the second site. An aliquot of initially processed samples was shipped to the other lab for a sample swap. Samples with "FQC" on the first run were re-tested.

Limit of Detection Testing:

Sample size: DNA samples from cell lines including BLMAsh (homozygous wild type, DD) - 4 samples/4 replicates; BLMAsh (heterozygous variant, DI) - 1 sample/8 replicates; BLMAsh (homozygous variant, II) - 1 sample/8 replicates.
Data source: Cell lines. Each DNA sample was tested at concentrations of 5, 15, and 50 ng/uL. Bi-directional Sanger sequencing was used to confirm BeadChip genotypes.
Annotation protocol: BeadChip genotyping was performed at two laboratory sites, with each site testing the same DNA sample replicates for each of 3 BLMAsh genotypes at 3 DNA concentrations, with 3 lots of reagents. BeadChip genotypes were compared with sequenced genotypes to determine correct call rates. Samples failing QC (call rate

Regulation Number and Section

§ 866.5940 Autosomal recessive carrier screening gene mutation detection system.

(a)
Identification. Autosomal recessive carrier screening gene mutation detection system is a qualitative in vitro molecular diagnostic system used for genotyping of clinically relevant variants in genomic DNA isolated from human specimens intended for prescription use or over-the-counter use. The device is intended for autosomal recessive disease carrier screening in adults of reproductive age. The device is not intended for copy number variation, cytogenetic, or biochemical testing.(b)
Classification. Class II (special controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 866.9, except § 866.9(c)(2). Autosomal recessive carrier screening gene mutation detection system must comply with the following special controls:(1) If the device is offered over-the-counter, the device manufacturer must provide 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.
(2) The device must use a collection device that is FDA cleared, approved, or classified as 510(k) exempt, with an indication for in vitro diagnostic use in DNA testing.
(3) The device's labeling must include a prominent hyperlink to the manufacturer's public Web site where the manufacturer shall make the information identified in this section publicly available. The manufacturer's home page, as well as the primary part of the manufacturer's Web site that discusses the device, must provide a prominently placed hyperlink to the Web page containing this information and must allow unrestricted viewing access. If the device can be purchased from the Web site or testing using the device can be ordered from the Web site, the same information must be found on the Web page for ordering the device or provided in a prominently placed and 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 would also have to be posted on the manufacturer's Web site. The information must include:
(i) A detailed device description including:
(A) Gene (or list of the genes if more than one) and variants the test detects (using standardized nomenclature, Human Genome Organization (HUGO) nomenclature, and coordinates).
(B) Scientifically established clinical validity of each variant detected and reported by the test, which must be well-established in peer-reviewed journal articles, authoritative summaries of the literature such as Genetics Home Reference (
http://ghr.nlm.nih.gov/ ), GeneReviews (http://www.ncbi.nlm.nih.gov/books/NBK1116/ ), or similar summaries of valid scientific evidence, and/or professional society recommendations, including:(
1 ) Genotype-phenotype information for the reported mutations.(
2 ) Relevant American College of Medical Genetics (ACMG) or American Congress of Obstetricians and Gynecologists (ACOG) guideline recommending testing of the specific gene(s) and variants the test detects and recommended populations, if available. If not available, a statement stating that professional guidelines currently do not recommend testing for this specific gene(s) and variants.(
3 ) Table of expected prevalence of carrier status in major ethnic and racial populations and the general population.(C) The specimen type (
e.g., saliva, whole blood), matrix, and volume.(D) Assay steps and technology used.
(E) Specification of required ancillary reagents, instrumentation, and equipment.
(F) Specification of the specimen collection, processing, storage, and preparation methods.
(G) 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.
(H) Information pertaining to the probability of test failure (
e.g., failed quality control) based on data from clinical samples, description of scenarios in which a test can fail (i.e., low sample volume, low DNA concentration, etc.), how customers will be notified, and followup actions to be taken.(I) Specification of the criteria for test result interpretation and reporting.
(ii) Information that demonstrates the performance characteristics of the device, including:
(A) Accuracy (method comparison) of study results for each claimed specimen type.
(
1 ) Accuracy of the device shall be evaluated with fresh clinical specimens collected and processed in a manner consistent with the device's instructions for use. If this is impractical, fresh clinical samples may be substituted or supplemented with archived clinical samples. Archived samples shall have been collected previously in accordance with the device's instructions for use, stored appropriately, and randomly selected. In some instances, use of contrived samples or human cell line samples may also be appropriate; the contrived or human cell line samples shall mimic clinical specimens as much as is feasible and provide an unbiased evaluation of the device's accuracy.(
2 ) Accuracy must be evaluated as compared to bidirectional sequencing or other methods identified as appropriate by FDA. Performance criteria for both the comparator method and device must be predefined and appropriate to the test's intended use. Detailed appropriate study protocols must be provided.(
3 ) Information provided shall include the number and type of specimens, broken down by clinically relevant variants, that were compared to bidirectional sequencing or other methods identified as appropriate by FDA. The accuracy, defined as positive percent agreement (PPA) and negative percent agreement (NPA), must be measured; accuracy point estimates must be greater than 99 percent (both per reported variant and overall) and uncertainty of the point estimate must be presented using the 95 percent confidence interval. Clinical specimens must include both homozygous wild type and heterozygous genotypes. The number of clinical specimens for each variant reported that must be included in the accuracy study must be based on the variant prevalence. Common variants (greater than 0.1 percent allele frequency in ethnically relevant population) must have at least 20 unique heterozygous clinical specimens tested. Rare variants (less than or equal to 0.1 percent allele frequency in ethnically relevant population) shall have at least three unique mutant heterozygous specimens tested. 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. Variants that have a point estimate for PPA or NPA of less than 99 percent (incorrect test results as 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. Point estimate of PPA should be calculated as the number of positive results divided by the number of specimens known to harbor variants (mutations) without “no calls” or invalid calls. The point estimate of NPA should be calculated as the number of negative results divided by the number of wild type specimens tested without “no calls” or invalid calls, for each variant that is being reported. Point estimates should be calculated along with 95 percent two-sided confidence intervals.(
4 ) Information shall be reported on the clinical positive predictive value (PPV) and negative predictive value (NPV) for carrier status (and where possible, for each variant) in each population. Specifically, to calculate PPV and NPV, estimate test coverage (TC) and the percent of persons with variant(s) included in the device among all carriers: PPV = (PPA * TC * π)/(PPA * TC * π + (1 − NPA) * (1 − π)) and NPV = (NPA * (1 − π))/(NPA (1 − π) + (1 − PPATC) * π) where PPA and NPA described either in paragraph (b)(3)(ii)(A)(4 )(i ) or in paragraph (b)(3)(ii)(A)(4 )(ii ) of this section and π is prevalence of carriers in the population (pre-test risk to be a carrier for the disease).(
i ) For the point estimates of PPA and NPA less than 100 percent, use the calculated estimates in the PPV and NPV calculations.(
ii ) Point estimates of 100 percent may have high uncertainty. If these variants are measured using highly multiplexed technology, calculate the random error rate for the overall device and incorporate that rate in the estimation of the PPA and NPA as calculated previously. Then use these calculated estimates in the PPV and NPV calculations. This type of accuracy study is helpful in determining that there is no systematic error in such devices.(B) Precision (reproducibility): Precision data must be generated using multiple instruments and multiple operators, on multiple non-consecutive days, and using multiple reagent lots. The sample panel must include specimens with claimed sample type (
e.g. saliva samples) representing different genotypes (i.e., wild type, heterozygous). 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. It must be clearly documented whether results were generated from clinical specimens, contrived samples, or cell lines. The study results shall state, in a tabular format, the variants tested in the study and the number of replicates for each variant, and what testing conditions were studied (i.e., number of runs, days, instruments, reagent lots, operators, specimens/type, etc). The study must include all nucleic acid 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 must be evaluated). The percentage of “no calls” or invalid calls, if any, in the study must be provided as a part of the precision (reproducibility) study results.(C) Analytical specificity data: Data must be generated evaluating the effect on test performance of potential endogenous and exogenous interfering substances relevant to the specimen type, evaluation of cross-reactivity of known cross-reactive alleles and pseudogenes, and assessment of cross-contamination.
(D) Analytical sensitivity data: Data must be generated demonstrating the minimum amount of DNA that will enable the test to perform accurately in 95 percent of runs.
(E) Device stability data: The manufacturer must establish upper and lower limits of input nucleic acid and sample stability that will achieve the claimed accuracy and reproducibility. Data supporting such claims must be described.
(F) Specimen type and matrix comparison data: Specimen type and matrix comparison data must be generated if more than one specimen type or anticoagulant can be tested with the device, including failure rates for the different specimen types.
(iii) If the device is offered over-the-counter, including cases in which the test results are provided direct-to-consumer, the manufacturer must conduct a study that assesses user comprehension of the device's labeling and test process and provide a concise summary of the results of the study. The following items must be included in the user study:
(A) The test manufacturer must perform pre- and post-test user comprehension studies to assess user ability to understand the possible results of a carrier test and their clinical meaning. The comprehension test questions must directly evaluate the material being presented to the user in the test reports.
(B) The test manufacturer must provide a carrier testing education module to potential and actual test report recipients. The module must define terms that are used in the test reports and explain the significance of carrier status.
(C) The user study must meet the following criteria:
(
1 ) The study participants must be comprised of a statistically justified and demographically diverse population (determined using methods such as quota-based sampling) that is representative of the intended user population. Furthermore, the users must be comprised of a diverse range of age and educational levels that have no prior experience with the test or its manufacturer. These factors shall 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 onsite 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 given to users must: Define the condition being tested and related symptoms; explain the intended use and limitations of the test; explain the relevant ethnicities regarding the variant tested; explain carrier status and relevance to the user's ethnicity; and provide links to additional information pertaining to situations where the user is concerned about their test results or would like followup information as indicated in test labeling. The study shall assess participants' ability to understand the following comprehension concepts: The test's limitations, purpose, and results.(
5 ) Study participants must be untrained, naive to the test subject of the study, and be provided only the materials that will be available to them when the test is marketed.(
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 to demonstrate that the education module and test reports are adequate for over-the-counter use.(D) A summary of the user comprehension study must be provided and include the following:
(
1 ) Results regarding reports that are provided for each gene/variant/ethnicity tested.(
2 ) Statistical methods used to analyze all data sets.(
3 ) Completion rate, non-responder rate, and reasons for non-response/data exclusion, as well as a summary table of comprehension rates regarding comprehension concepts (purpose of test, test results, test limitations, ethnicity relevance for the test results, etc.) for each study report.(4) Your 21 CFR 809.10 compliant labeling and any test report generated must include the following warning and limitation statements, as applicable:
(i) A warning that reads “The test is intended only for autosomal recessive carrier screening in adults of reproductive age.”
(ii) A statement accurately disclosing the genetic coverage of the test in lay terms, including, as applicable, information on variants not queried by the test, and the proportion of incident disease that is not related to the gene(s) tested. For example, where applicable, the statement would have to include a warning that the test does not or may not 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 be disease-related. Or, where applicable, the statement would have to include a warning that the basis for the disease for which the genetic carrier status is being tested is unknown or believed to be non-heritable in a substantial number of people who have the disease, and that a negative test result cannot rule out the possibility that any offspring may be affected with the disease. The statement would have to include any other warnings needed to accurately convey to consumers the degree to which the test is informative for carrier status.
(iii) For prescription use tests, the following warnings that read:
(A) “The results of this test are intended to be interpreted by a board-certified clinical molecular geneticist or equivalent and should be used in conjunction with other available laboratory and clinical information.”
(B) “This device is not intended for disease diagnosis, prenatal testing of fetuses, risk assessment, prognosis or pre-symptomatic testing, susceptibility testing, or newborn screening.”
(iv) For over-the-counter tests, a statement that reads “This test is not intended to diagnose a disease, or tell you anything about your risk for developing a disease in the future. On its own, this test is also not intended to tell you anything about the health of your fetus, or your newborn child's risk of developing a particular disease later on in life.”
(v) For over-the-counter tests, the following warnings that read:
(A) “This test is not a substitute for visits to a healthcare provider. It is recommended that you consult with a healthcare provider if you have any questions or concerns about your results.”
(B) “The test does not diagnose any health conditions. Results should be used along with other clinical information for any medical purposes.”
(C) “The laboratory may not be able to process your sample. The probability that the laboratory cannot process your saliva sample can be up to [actual probability percentage].”
(D) “Your ethnicity may affect how your genetic health results are interpreted.”
(vi) For a positive result in an over-the-counter test when the positive predictive value for a specific population is less than 50 percent and more than 5 percent, a warning that reads “The positive result you obtained may falsely identify you as a carrier. Consider genetic counseling and followup testing.”
(vii) For a positive result in an over-the-counter test when the positive predictive value for a specific population is less than 5 percent, a warning that reads “The positive result you obtained is very likely to be incorrect due to the rarity of this variant. Consider genetic counseling and followup testing.”
(5) The testing done to comply with paragraph (b)(3) of this section must show the device meets or exceeds each of the following performance specifications:
(i) The accuracy must be shown to be equal to or greater than 99 percent for both PPA and NPA. Variants that have a point estimate for PPA or NPA of less than 99 percent (incorrect test results as compared to bidirectional sequencing or other methods identified as appropriate by FDA) must not be incorporated into test claims and reports.
(ii) Precision (reproducibility) performance must meet or exceed 99 percent for both positive and negative results.
(iii) The user comprehension study must obtain values of 90 percent or greater user comprehension for each comprehension concept.
(6) The distribution of this device, excluding the collection device described in paragraph (b)(2) of this section, shall be limited to the manufacturer, the manufacturer's subsidiaries, and laboratories regulated under the Clinical Laboratory Improvement Amendments.

Summary

EVALUATION OF AUTOMATIC CLASS III DESIGNATION FOR The 23andMe Personal Genome Service Carrier Screening Test for Bloom Syndrome

DECISION SUMMARY

This decision summary corrects the decision summary dated February 2015.

A. DEN Number:

DEN140044

B. Purpose for Submission:

De Novo request for evaluation of automatic class III designation for the 23andMe Personal Genome Service (PGS) Carrier Screening Test for Bloom Syndrome

C. Measurands:

Genomic DNA obtained from a human saliva sample

D. Type of Test:

The 23andMe PGS Carrier Screening Test for Bloom Syndrome, using the Illumina Infinium BeadChip (23andMe BeadChip), is designed to be capable of detecting specific single nucleotide polymorphisms (SNPs) as well as other genetic variants. The 23andMe PGS Carrier Screening Test for Bloom Syndrome is a molecular assay indicated for use for the detection of the BLMAsh variant in the BLM gene from saliva collected using the OrageneDx® saliva collection device (OGD-500.001). Results are analyzed using the Illumina iScan System and Genome Studio and Coregen software. The 23andMe PGS Carrier Screening Test for Bloom Syndrome can be used to determine carrier status for Bloom syndrome, but cannot determine if a person has two copies of the BLMAS variant.

E. Applicant:

23andMe, Inc.

F. Proprietary and Established Names:

23andMe Personal Genome Service Carrier Screening Test for Bloom Syndrome

G. Regulatory Information:

1. Regulation section:
- 21 CFR 866.5940
1. Classification:

Class II

1. Product code(s):
  PKB
1. Panel:
  82- Immunology

H. Indication(s) for use:

1. Indication(s) for use:
  The 23andMe PGS Carrier Screening Test for Bloom Syndrome is indicated for the detection of the BLMAsh variant in the BLM gene from saliva collected using an FDA cleared collection device (Oragene DX model OGD-500.001). This test can be used to determine carrier status for Bloom syndrome in adults of reproductive age, but cannot determine if a person has two copies of the BLM140 variant. The test is most relevant for people of Ashkenazi Jewish descent.

2. Special conditions for use statement(s):

For over-the-counter (OTC) use.
This test is not intended to diagnose a disease, to tell you anything about the health of your fetus, or your risk or your new born child's risk of developing a particular disease later on in life.
This test is not a substitute for visits to a healthcare provider. It is recommended that you consult with a healthcare provider if you have any questions or concerns about your results.
23andMe PGS Carrier Screening Test for Bloom Syndrome does not detect all genetic variants associated with Bloom Syndrome. The absence of a variant tested does not rule out the presence of other genetic variants that may be disease related.
The test is intended only for autosomal recessive carrier screening in adults of reproductive age.
The test does not diagnose any health conditions. Results should be used along with other clinical information for any medical purposes.
The laboratory may not be able to process a patient's sample. The probability that the laboratory cannot process the sample can be up to 7.6%.
A user's ethnicity may affect how the genetic test results are interpreted.

1. Subject to meeting the limitations contained in the special controls under regulation 21 CFR 866.5940.

4. Special instrument requirements:

The 23andMe PGS Carrier Screening Test for Bloom Syndrome is to be performed using the Tecan Evo and Illumina iScan instruments.

GenomeStudio is a modular software application that is used to view and analyze genotypic data obtained from the iScan. Coregen software conducts a variety of control checks on the file, resulting in a final analytical genotype profile for each sample. The data is used to generate test reports that are based on information from reported scientific findings on genotypes.

I. Device Description:

genotypic data obtained from the iScan.

J. Substantial Equivalence Information:

1. Predicate device name(s):
  No predicate device exists.
1. Predicate 510(k) number(s):
  Not applicable.
1. Comparison with predicate:
  Not applicable.

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

Not applicable.

L. Test Principle:

The PGS is indicated to be performed using the BeadChip v4 assay (Illumina HumanOmniExpress-24 format chip), which covers ~750,000 SNPs. The BeadChip 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 SNP 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/processing and BeadChip quantification and scanning, respectively. The genotype content is separated, analyzed, and then integrated into pre-defined report templates specific for each condition associated with each genotype. The iScan software uses the dmap file to associate signal intensity measured by the iScan Reader with bead type. The algorithm uses sequential hybridizations of dye-labeled oligonucleotides, or decoders, complementary to bead sequences to create a combinatorial decoding scheme for arravs. The approach uses longer sequences; each is designed to hybridize to a defined target with high specificity. It is capable of decoding, with high accuracy, many thousands of bead types. Each bead type is defined by a unique DNA sequence that is recognized by a complementary decoder. Genotypes are determined using the GenomeStudio software package.

The PGS is indicated to detect the BLMAsh variant. The BLMAsh variant results from a deletion of six letters and an insertion of seven letters in the sequence of the gene. It results in a shortened protein that does not work properly (Typical Sequence → Deletion and Insertion > TAGATTC Variant Sequence).

Intensity values that fall outside of the expected range of intensities are returned as "no-calls"; however, all homozygous variant (BLMAsh II) genotype samples receive a "no call" result, since the calling software was designed not to detect BLMdsb II homozygous variant genotypes. Such results will be reported as "could not be determined" to the customer in their test report. Genetic results are returned to the customer in a secure online account on the 23andMe website.

M. Performance Characteristics:

Saliva samples were collected using the Oragene Dx saliva collection device (OGD-500.001). The samples were tested on the Illumina Infinium BeadChip. Results were analyzed using the Illumina iScan System and GenomeStudio and Coregen software.

1. Analytical performance:

a. Precision/Reproducibility

For evaluation of precision/reproducibility, two studies were conducted: the first study was conducted with human cell line samples and a second study was conducted with human saliva samples.

Precision Study with Human Cell Line Samples

The precision study was conducted at the b(4) TS/CCI Six DNA samples taken from b(4) TS/CCI cell b(4) TS/CCI lines (4 BLM homozygous common ("DD") samples, 1 BLMAs" heterozygous ("DI") sample and 1 BLMAsh homozygous rare ("II") sample) were tested over 5 days by 3 operator teams at site 1 and 33 operator teams at site 2. To confirm the BeadChip genotype, each [4] TS/CC sample was also sequenced by bi-directional Sanger Sequencing. Samples were tested with 4 lots of reagents (lot 1, lot 2 and lot 3 at site 1 and lot 2, lot 3 and lot 4 at site 2) using 3 Tecan instruments and 3 iScan instruments in different combinations. For each of 4 "DD" samples, there were 36 replicates per day per laboratory site with total number of 360 replicates (36 x 5 x 2). The other two samples, "DI" and "II", had 72 replicates per day per site with total number of 720 replicates (72 x 5 x 2). Information regarding samples that failed quality control (FQC) was also evaluated.

Results of the Study Stratified by Site are presented below:

| Site 1 | Total number
of replicates | Number of
correct calls | Number of
miscalls | Number of
"FQC"
replicates | Percent of
"FQC"
replicates |
|------------------|-------------------------------|----------------------------|-----------------------|----------------------------------|-----------------------------------|
| Sample 1
"DD" | 180 | 173 | 0 | 7 | 3.89% |
| Sample 2
"DD" | 180 | 174 | 0 | 6 | 3.33% |
| Sample 3 | 180 | 175 | 0 | 5 | 2.78% |

"DD"
Sample 4
"DD"	180	175	0	5	2.78%
Sample "DI"	360	350	0	10	2.78%
Sample "II"	360	343	0	17	4.72%
Total	1,440	1,390	0	50	3.47%

| Site 2 | Total number
of replicates | Number of
correct calls | Number of
miscalls | Number of
"FQC"
replicates | Percent of
"FQC"
replicates |
|------------------|-------------------------------|----------------------------|-----------------------|----------------------------------|-----------------------------------|
| Sample 1
"DD" | 180 | 171 | 0 | 9 | 5.00% |
| Sample 2
"DD" | 180 | 174 | 0 | 6 | 3.33% |
| Sample 3
"DD" | 180 | 176 | 0 | 4 | 2.22% |
| Sample 4
"DD" | 180 | 179 | 0 | 1 | 0.56% |
| Sample "DI" | 360 | 350 | 0 | 10 | 2.78% |
| Sample "II" | 360 | 350 | 0 | 10 | 2.78% |
| Total | 1,440 | 1,400 | 0 | 40 | 2.78% |

Results of the study (percent of "FQC" replicates) stratified by instrument combinations are shown for each site below:

Percent of "FQC" for nine different combinations of 3 Tecan and 3 iScan instruments at Percent
Site

		Percent of "FQC"
Tecan	iScan	4 Samples
"DD"	Sample "DI"	Sample "II"	Total
1	1	0.00%
(0/96)	0.00%
(0/48)	0.00%
(0/48)	0.00%
1	2	0.00%
(0/48)	4.17%
(1/24)	4.17%
(1/24)	2.08%
1	3	8.33%
(8/96)	4.17%
(2/48)	2.08%
(1/48)	5.73%
2	1	0.00%
(0/96)	0.00%
(0/48)	0.00%
(0/48)	0.00%
2	2	7.29%
(7/96)	4.17%
(2/48)	14.58%
(7/48)	8.33%
2	3	4.17%
(2/48)	12.50%
(3/24)	12.50%
(3/24)	8.33%
3	1	4.17%
(2/48)	0.00%
(0/24)	0.00%
(0/24)	2.08%
3	2	0.00%
(0/48)	0.00%
(0/48)	0.00%
(0/48)	0.00%

3	3	4.17%	4.17%	10.42%
		(4/96)	(2/48)	(5/48)	5.73%
Percent of
"FQC" for
nine different
combinations
of 3 Tecan		Percent of "FQC"
and 3 iScan
instruments at
b(4) TS/CCl
Tecan	iScan	4 Samples
"DD"	Sample "DI"	Sample "II"	Total
4	4	4.17%	0.00%	4.17%	3.13%
		(4/96)	(0/48)	(2/48)
4	5	2.08%	2.08%	6.25%	3.13%
		(2/96)	(1/48)	(3/48)
4	6	0.00%	0.00%	0.00%	0.00%
		(0/48)	(0/24)	(0/24)
5	4	0.00%	5.36%	0.00%	1.34%
		(0/112)	(3/56)	(0/56)
5	5	6.25%	0.00%	0.00%	3.13%
		(3/48)	(0/24)	(0/24)
5	6	12.5%	7.50%	2.50%	8.75%
		(10/80)	(3/40)	(1/40)
6	4	0.00%	0.00%	0.00%	0.00%
		(0/48)	(0/24)	(0/24)
6	5	1.04%	4.17%	4.17%	2.60%
		(1/96)	(2/48)	(2/48)
6	6	0.00%	2.08%	4.17%	1.56%
		(0/96)	(1/48)	(2/48)

The percent of FQC for 9 combinations of Tecan and iScan instruments at "frises ranged from 0% to 8.33%. The percent of FQC for 9 combinations of Tecan and
iScan instruments at 7(4) TSCCI ranged from 0% to 8.75%

Results of the Study (percent of "FQC" replicates) Stratified by Lot of Reagents are presented below:

| | Sample 1
"DD" | Sample 2
"DD" | Sample 3
"DD" | Sample 4
"DD" | Sample
"DI" | Sample
"II" | Total |
|-------|------------------|------------------|-------------------|------------------|------------------|-------------------|--------------------------|
| Lot 1 | 8.33%
(5/60) | 8.33%
(5/60) | 0.00%
(0/60) | 0.00%
(0/60) | 5.00%
(6/120) | 8.33%
(10/120) | 5.42%
(26/480) |
| Lot 2 | 5.00%
(6/120) | 0.83%
(1/120) | 2.50%
(3/1200) | 0.00%
(0/120) | 1.67%
(4/240) | 3.75%
(9/240) | 2.40%
(23/960) |
| Lot 3 | 4.17%
(5/120) | 4.17%
(5/120) | 5.00%
(6/120) | 4.17%
(5/120) | 1.67%
(4/240) | 1.67%
(4/240) | 3.02%
(29/960) |

Lot 4	0.00%	1.67%	0.00%	1.67%	5.00%	3.33%	2.50%
	(0/60)	(1/60)	(0/60)	(1/60)	(6/120)	(4/120)	(12/480)

The percent of "FQC" replicates for 4 different lots of reagents ranged from 2.4% to 5.4%.

The combined data of the reproducibility study for 6 human cell line samples are presented in the table below:

| | Total number
of replicates | Number of
correct calls | Number of
miscalls | Number of
"FQC"
replicates | Percent of
"FQC"
replicates |
|------------------|-------------------------------|----------------------------|-----------------------|----------------------------------|-----------------------------------|
| Sample 1
"DD" | 360 | 344 | 0 | 16 | 4.44% |
| Sample 2
"DD" | 360 | 348 | 0 | 12 | 3.33% |
| Sample 3
"DD" | 360 | 351 | 0 | 9 | 2.50% |
| Sample 4
"DD" | 360 | 354 | 0 | 6 | 1.67% |
| Sample "DI" | 720 | 700 | 0 | 20 | 2.78% |
| Sample “II” | 720 | 693 | 0 | 27 | 3.75% |
| Total | 2,880 | 2,790 | 0 | 90 | 3.13% |

96.9% (2,790/2,880) replicates produced correct genotyping results and 3.1% (90/2,880) replicates did not pass Quality Control (QC) acceptance criteria. Samples with failed QC on the first run are re-tested per laboratory SOPs; therefore, an anticipated rate of samples with two times failed QC based on precision study data of the human cell line samples is 0.1% (=0.0313 x 0.0313).

Laboratory Reproducibility Study with Saliva Samples

A reproducibility study was performed at the same 2 sites as the reproducibility study with human cell line samples with a total of 105 BLMAsh homozygous common ("DD") saliva samples obtained from individuals using the 23andMe Saliva Collection kit (Oragene-DX, OGD500.001, saliva collection kit). Sample processing was performed at both sites and tested with PGS test for Bloom syndrome. Fifty samples were initially processed at the first site and 55 samples were processed at the second site. A sample swap was performed, where an aliquot of the initially processed samples were shipped to the other lab. Results for 105 saliva samples are presented below for both sites.

		Saliva samples “DD”
		Site 1	Site 2
PGS
Carrier
test
for
Bloom
Syndrome	First run
"1 Variant Detected"	0	0
	First run
"0 Variants Detected"	104	87
First
run
FQC
Result	Re-run
"1 Variant Detected"	0	0
	Re-run
"0 Variants Detected"	1	10
	Re-run
FQC	0	8
Total		105	105

Laboratory Reproducibility Study with First Run and Re-run Summary Results

Since the only samples tested were homozygous common (DD), there were no results for the "1 Variant Detected" category for either site. The percent of saliva samples with "FOC" on the first run was 1.0% (1/105) at site 1 and 17.1% (18/105) at site 2. Samples with the "FQC" on the first run were re-tested and the percent of saliva samples with a final failed QC result (Re-run FQC) was 0% (0/105) at site 1 and 7.6% (8/105) at site 2.

b. Linearity/assay Reportable Range:
Not applicable
c. Traceability, Stability, Expected Values (controls, calibrators, or methods):
The PGS requires two types of controls, the sample processing control and the reproducibility control. The sample processing control material is generated from b(4) TS/CCl

the 23andMe BeadChip according to routine Standard Operating Procedures (SOPs) at the contracted laboratory sites.

The reproducibility control consists of b(4) TS/CCI

Each new lot of the

reproducibility control is tested by comparison with reference BeadChip genotype results. DNA is extracted from this cell suspension and genotyped using the 23andMe BeadChip, according to routine SOPs at the contracted laboratory.

The sample processing control is routinely run on every sample genotyping plate and the reproducibility control is routinely run approximately once per week. Historical data from all such runs were analyzed for one lot of the sample processing control spanning 3 months and one lot of the reproducibility control spanning 1 year.

Stability protocols and acceptance criteria were reviewed and deemed acceptable. The information provided demonstrates that the sample processing control is stable for up to 3 months and the reproducibility control is stable for up to 12 months.

d. Detection Limit:
Limit of Detection testing was performed using DNA samples [4) Tscc cell lines. The following samples/genotypes/replicates were tested: BLMAsh (homozygous wild type, DD), 4 samples/4 replicates per sample; BLMAsh (heterozygous variant, DI), 1 sample/8 replicates per sample; BLMAsh (homozygous variant, II), 1 sample/8 replicates per sample. Each DNA sample was tested at the following concentrations: 5, 15, and 50 ng/μL. BeadChip genotyping was performed [4] Tscc] and b(4) TS/CCI DNA, where each site tested the same DNA sample replicates for each of 3 BLMAsh genotypes at 3 DNA concentrations, with 3 lots of reagents. To confirm the BeadChip genotype, each 64756Cl sample was also sequenced by bi-directional Sanger sequencing. BeadChip genotypes were compared with sequenced genotypes to determine the rates of correct BeadChip genotype calls at each DNA concentration. If a sample replicate failed BeadChip or sequencing Quality Control (OC) criteria, it was marked as "FQC" ("failed QC") if the sample replicate did not demonstrate a call rate ≥ 0.980.

The lower LoD was defined as the lowest DNA concentration at which at least 95% of samples yielded the correct call at each of two laboratory sites. The LoD study vielded 100% correct call rates for all samples across all reagent lots, at all sample concentrations tested at two independent laboratory sites. Therefore, the study passed the acceptance criteria of 95% correct calls at the lowest concentration tested (5 ng/uL). The performance requirement for the PGS, has been set at a minimum of 15 ng/uL DNA and maximum of 50 ng/uL DNA.

Results from the studies also indicated that all but one replicate passed OC acceptance criteria on the re-run (per laboratory SOP) and yielded correct genotype calls. One replicate failed QC (due to call rate 0.980". Upon re-running these samples, all replicates produced correct genotype calls.

f. Assay Cut-off:
Not applicable.
Specimen Stability at 2-8° C g.
Saliva samples for testing are collected with the Oragene Dx OGD-500.001 collection device. See K141410 for sample stability information.
h. Shipping Stability
Saliva samples are shipped for testing in the Oragene Dx OGD-500.001 collection device. See K141410 for sample shipping stability information.
1. Comparison Studies:
- Method Comparison with Predicate Device: a.

Accuracy was evaluated by the agreement of the genetic variant determinations by this test with bi-directional sequencing results.

Saliva samples were randomly selected from the 23andMe Biobank, in which saliva samples were collected using the DNA Genotek OrageneDx 500.001 collection device. Saliva sample selection was blinded to previously determined genotypes and at least 20 carrier samples (DI, as detected by genotyping) were selected. A total of 65 saliva samples were selected for the study (25 DD and 22 DI samples tested at Site 1; 18 DI samples samples tested at Site 2) in addition to 6 human cell line samples tested (4 DD samples, 1 DI sample and 1 II sample tested at both sites). All 71 samples were sequenced using bi-directional sequencing. The comparison study was conducted at 2 sites; results of the test were compared with sequencing results. If a replicate fails QC ("FQC") criteria on the first run, the replicate was re-run once using the same sample. Five saliva samples failed QC at Site 1 and 1 saliva sample failed QC at the Site 2. All re-run samples produced correct genotype results. Study

Summary of Comparison Data of Saliva Samples for b(4) TS/CCI
			Bi-directional sequencing
			"DI"	"DD"
PGS
Carrier
test
for
Bloom
Syndrome	First run
"1 Variant Detected"		21	0
	First run
"0 Variants Detected"		0	21
First
Run
FQC
Result	Re-run
"1 Variant
Detected"	1	0


	Re-run
"0 Variants
Detected"	0	4

	Re-run
FQC		0	0
Total			22	25

results and % agreement are provided in the tables below:

Summary of Comparison Data of Saliva Samples for b(4) TS/CCI

		Bi-directional sequencing
		"DI"	"DD"
PGS
Carrier
test
for
Bloom
Syndrome	First run	"1 Variant Detected"	17	0
		"0 Variants Detected"	0	0
	First
Run
FQC
Result	Re-run
"1 Variant
Detected"	1	0
		Re-run
"0 Variants
Detected"	0	0
		Re-run
FQC	0	0
		Total

The homozygous common (DD) genotype results at both sites did not produce results for the "1 Variant Detected Category" and vice versa for the heterozygous variant genotype regarding the "0 Variants Detected" category. All samples that failed QC on the first run produced correct genotype results upon re-running them at both sites.

| | Positive Percent Agreement
Saliva Samples with “DI” by
bi-directional sequencing | | Negative Percent Agreement
Saliva Samples with “DD” by
bi-directional sequencing | |
|----------|----------------------------------------------------------------------------------------|--------------------------|----------------------------------------------------------------------------------------|--------------------------|
| | Percent correct
results | %FQC
on the first run | Percent correct
results | %FQC
on the first run |
| Site 1 | 100% (22/22) | 4.5% (1/22) | 100% (25/25) | 16.0% (4/25) |
| Site 2 | 100% (18/18) | 5.6% (1/18) | n/a | n/a |
| Combined | 100% (40/40) | 5.0% (2/40) | 100% (25/25) | 16.0% (4/25) |

Positive and Negative Percent Agreements for Saliva Samples for Both Sites

Results for the cell line samples tested at both sites are as follows:

i) The DI sample had correct genotyping results on the first run at both sites;
ii) All 4 DD samples had correct genotyping on the first run at both sites;

iii) The II sample failed QC on the first run at Site 2 and produced a correct (no call) result upon re-running the sample.

The following table presents PPA and NPA for saliva and human cell line samples combined.

| | Percent of
correct calls | 95% CI |
|-------------------------------------|-----------------------------|-----------------|
| Positive Percent
Agreement (PPA) | 100% (41/41) | 91.4% to 100%* |
| Negative Percent
Agreement (NPA) | 100% (29/29) | 88.3% to 100%* |
| Overall Agreement | 100% (70/70) | 96.3% to 100%** |

PPA and NPA for Saliva and Human Cell Line Samples Combined for Both Sites

*95% two-sided confidence interval

** 95% one-sided confidence interval

Overall agreement was 100% (70/70) with 95% confidence interval of 96.3% to 100%.

b. Matrix Comparison:
Not applicable. This test is for use with human saliva samples only.
1. Clinical Studies:

Clinical Performance

The BLMAsh variant covered by this test is mainly found in people of Ashkenazi Jewish descent. Approximately 1 in 107 people (0.93%) with this ethnicity carries this variant. The BLMAsh variant is rare and not well studied in other ethnic groups.

Ancestry Group	Frequency	Number of Tested
Ashkenazi Jewish	1.03%	b(4) TS/CC
European	0.02%
Latino	99%1	Unknown
Pre-test carrier risk	1 in 1071	Likely 0.950.

Reproducibility Control (RC) evaluates performance at weekly ● intervals from extraction through detection. RC consists of the same b(4) TS/CCl as the SP control, but is assayed from b(4) TS/CCI

P. Other Supportive Instrument Performance Characteristics Data Not Covered In The "Performance Characteristics" Section above:

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

O. Identified Risks and Required Mitigations:

Carrier screening is a type of genetic testing performed on people who display no symptoms for a recessive genetic disorder but may be at risk for passing it on to their children. A carrier for a genetic disorder has inherited one normal and one abnormal allele for a gene associated with the disorder. A child must inherit two abnormal alleles in order for symptoms to appear.

By definition, autosomal (non-sex chromosome-related) recessive disorders require that two abnormal copies of a gene, one inherited from each parent, be present in order for the disorder to be manifested. Therefore, to have a child with an autosomal recessive disorder both parents must be carriers of an abnormal gene copy. When both parents are carriers for the abnormal copy, there is a 1 in 4 chance (25%) that the child will inherit two abnormal copies of the gene and manifest the specific disease/condition or trait.

Most autosomal recessive genetic diseases are very rare with frequencies much less than 1% in the general population. Some autosomal recessive genetic diseases are more common in certain ethnic groups; however, even in these ethnic groups disease frequencies tend to be very low.

In defining an inheritance pattern of genetic disease, one must demonstrate the appropriate genetic patterns are present in an informed population that includes affected persons. Reputable iournals and professional bodies would be very likely to reject a weak or mistaken

interpretation of the mode of inheritance of a genetic disease, because in order to demonstrate heritability, certain criteria must be met, e.g., a family with affected and unaffected individuals would have to display the expected segregation of alleles, or the inheritance pattern would be considered inconclusive. There is very little danger that fraudulent or wishful claims for autosomal recessive inheritance would be accepted by such groups.

Risk of False Results:

When considering the risks of tests for being a carrier of an abnormal gene for an autosomal recessive genetic disease, one should consider the effects of both false positive and false negative results, as well as the appropriate mitigations for these risks. Note that the risks of carrier testing are generally similar regardless of the genetic carrier condition to be detected.

Because carrier screening is only intended to detect heterozygotes (carriers) and not homozygotes, false positive results would suggest that a person was a carrier of a mutation, but would not generally contain any information that could lead to conclusions of disease for the tested person. Further, no conclusion about an individual's future children could be made given the contribution of the carrier status of the child's second parent would be needed for such a conclusion.
. As stated above, false positive results obtained by one individual would have to be complemented by a partner who was also a carrier to cause a couple to consider action based upon the test result. In this scenario, the false-positive could lead to couples choosing not to get married nor have children, or could lead to unnecessary fetal testing in current or future pregnancies. Fetal testing may consist of amniocentesis or chorionic villus sampling (CVS), which pregnant women might already be scheduled for due to other risk factors such as age. Amniocentesis and CVS do carry a risk of spontaneous abortion, so thev are not risk-free themselves.
The probability of two false-positive carrier results for a couple is expected to be . significantly smaller than for one false-positive.
A false-positive result for an individual could potentially lead to adverse . psychological effects, particularly if that individual didn't fully understand the nature of autosomal recessive disorders (i.e. that both the mother and father must both be carriers in order to have a 25% chance that their child would have the disorder).

Risk of False-Negative Results:

False negative results would suggest that a person was not a carrier, but would not directly affect the health of the tested person.
. The risk of a clinical false-negative result is already "high" because not all clinically relevant mutations are known or tested for most diseases. The number of people who are true carriers who would be detected by any test is known as the test's "coverage". The clinical false-negative rate due to "coverage" less than 100% is likely higher than the false-negative rate from analytical failure or random error of a test.
. Current genetic testing recommendations typically recommend that initially only one member of a couple be tested for carrier status; therefore, the risks associated with false-negative results generally occur when only one member of a couple is tested and experiences a false-negative result. The risk of the false-negative would only have consequence if the non-tested partner was a carrier of the condition or disorder. In this case, there is a 25% chance that a future child would inherit the condition or disorder.

Additional Risks:

In addition to the risks associated with false-positive or negative results, we identified an additional risk associated with this device system, which is the risk that results from an incorrect interpretation of the test result. This could be characterized by an individual not understanding the nature of autosomal recessive carrier screening tests (i.e. that both the mother and father must both be carriers in order to have a 25% chance that their child would have the disorder) and making critical decisions based upon this information. The risk for this element is considered to be greater for over-the-counter devices where no healthcare professional is directly involved with test ordering or interpretation.

Special Controls:

The special controls outlined in the Order address the risks identified above:

Special control 1 mandates that OTC manufacturers of these tests must provide . information to a potential or actual test report recipient about how to obtain access to a board-certified clinical molecular geneticist or equivalent to assist in pre and posttest counseling to a potential purchaser and actual test report recipient.
Special control 2 requires the use of a collection device that is FDA . cleared/approved or classified as 510(k) exempt, with an indication for use in in vitro diagnostic use in DNA testing. The use of a FDA-compliant collection device provides assurances regarding safety, effective and quality of that component, which helps assure safety and effectiveness of the test system.
. Special control 3 includes a detailed outline of clinical and analytical and performance information that must be generated and posted on the manufacturer's website. This special control also provides details on how analytical testing must be performed and provides criteria on the appropriate standard for performance for many

of these elements. This mitigates risk through lowering the probability of inaccurate test results and provides transparency on test limitations and performance.

Special control 4 outlines what the test should and should not be used for and . mandates appropriate warning statements be included in product labeling. These controls help ensure that users have the information available to enable them to understand the limitations of the test results prior to ordering and after receiving results. The controls also provide context for the use and further interpretation of the results.
Special control 5 provides specific requirements for accuracy and precision/reproducibility performance characteristics. Providing such specific requirements mitigates risk through lowering the probability of inaccurate test results by ensuring a minimum level of performance.
. Special control 6 limits the distribution of devices, excluding the collection device to the manufacturer, manufacturer's subsidiaries and laboratories subject to regulation under the Clinical Laboratory Improvement Amendments. This limitation is intended to mitigate risk through lowering the probability of inaccurate test results by ensuring that testing is performed by qualified individuals and in a manner that provides greater assurance of quality of the testing process.

Identified Risks	Required Mitigations
Incorrect understanding of the device and test system	Special Controls (1) and (4)
Incorrect test results	Special Controls (2), (3), (5) and (6)
Incorrect interpretation of test results	Special Controls (1), (3), (4), and (5)

Identified Risks and Required Mitigations Table R

S. Benefit/Risk Analysis:

Summary
Summary of
the Benefit(s)	People ≥ 18 years of age may be able to determine preconception carrier status
to assist in reproductive decision making.

| Summary of
the Risk(s) | Associated device risks include erroneous false negative results due to device or
user error or false positive results due to device or user error. A person with a
false positive result could have unnecessary additional testing (e.g., invasive
prenatal testing with risk of miscarriage) or make an inappropriate reproductive
choice (e.g., deciding against pregnancy due to unwarranted concern for having
an affected child). A person with a false negative result could forego prenatal
testing and/or have a child with an autosomal recessive disorder. |
|----------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Conclusions
Do the
probable
benefits
outweigh the
probable risks? | Based upon the nature of autosomal recessive carrier testing, which requires
that both parents carry a mutation in order to have a 25% chance of bearing an
affected child, and the level of scientific evidence required to establish
autosomal recessive inheritance patterns which are straightforward to verify, in
conjunction with the special controls drafted for this class of test systems, OIR
determines that the probable benefits of Autosomal Recessive Carrier Screening
Gene Mutation Detection Systems outweigh their probable risks. |

T. Conclusion:

The information provided in this de novo submission is sufficient to classify this device into class II under regulation 21 CFR 866.5940. FDA believes that special controls, along with the applicable general controls, provide reasonable assurance of the safety and effectiveness of the device type. The device is classified under the following:

Product Code: РКВ

Autosomal Recessive Carrier Screening Gene Mutation Detection System. Device Type:

Class: II (special controls).

Regulation: 21 CFR 866.5940

(a)Identification. Autosomal recessive carrier screening gene mutation detection system is a qualitative in vitro molecular diagnostic system used for genotyping of clinically relevant variants in genomic DNA isolated from human specimens intended for prescription use or overthe-counter use. The device is intended for autosomal recessive disease carrier screening in adults of reproductive age. The device is not intended for copy number variation, cytogenetic, or biochemical testing.
(b) Classification. Class II (special controls). Autosomal recessive carrier screening gene mutation detection system must comply with the following special controls:

(1) If the device is offered over-the-counter, the device manufacturer must provide 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.
(2) The device must use a collection device that is FDA cleared, approved, or classified as 510(k) exempt, with an indication for in vitro diagnostic use in DNA testing.
(3) The device's labeling must include a prominent hyperlink to the manufacturer's public website where the manufacturer shall make the information identified in this subsection 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 prominently-placed 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 prominently-placed and 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 would also have to be posted on the manufacturer's website. The information must include:
- A detailed device description including: (i)
  - (A)Gene (or list of the genes if more than one) and variants the test detects (using standardized nomenclature, Human Genome Organization (HUGO) nomenclature and coordinates).
  - (B) Scientifically established clinical validity of each variant detected and reported by the test, which must be well-established in peer-reviewed journal articles, authoritative summaries of the literature such as Genetics Home Reference (http://ghr.nlm.nih.gov/), GeneReviews (http://www.ncbi.nlm.nih.gov/books/NBK1116/) or similar summaries of valid scientific evidence, and/or professional society recommendations, including:
    - (1) Genotype-phenotype information for the reported mutations.
    - (2) Relevant American College of Medical Genetics (ACMG) or American Congress of Obstetricians and Gynecologists (ACOG) guideline recommending testing of the specific gene(s) and variants the test detects and recommended populations, if available. If not available, a statement stating that professional guidelines currently do not recommend testing for this specific gene(s) and variants.
    - (3) Table of expected prevalence of carrier status in major ethnic and racial populations and the general population.
  - (C) The specimen type (e.g., saliva, whole blood), matrix and volume.
  - (D) Assay steps and technology used.
  - (E) Specification of required ancillary reagents, instrumentation and equipment.

(F) Specification of the specimen collection, processing, storage and preparation methods.
(G)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.
(H) Information pertaining to the probability of test failure (e.g., failed quality control) based on data from clinical samples, description of scenarios in which a test can fail (i.e., low sample volume, low DNA concentration, etc.), how customers will be notified and follow-up actions to be taken.
(I) Specification of the criteria for test result interpretation and reporting.
(ii) Information that demonstrates the performance characteristics of the device, including:
- (A)Accuracy (method comparison) of study results for each claimed specimen type.
  - (1) Accuracy of the device shall be evaluated with fresh clinical specimens collected and processed in a manner consistent with the device's instructions for use. If this is impractical, fresh clinical samples may be substituted or supplemented with archived clinical samples. Archived samples shall have been collected previously in accordance with the device's instructions for use, stored appropriately and randomly selected. In some instances, use of contrived samples or human cell line samples may also be appropriate; the contrived or human cell line samples shall mimic clinical specimens as much as is feasible and provide an unbiased evaluation of the device's accuracy.
  - (2) Accuracy must be evaluated as compared to bidirectional sequencing or other methods identified as appropriate by FDA. Performance criteria for both the comparator method and device must be pre-defined and appropriate to the test's intended use. Detailed appropriate study protocols must be provided.
  - (3) Information provided shall include the number and type of specimens, broken down by clinically relevant variants, that were compared to bidirectional sequencing or other methods identified as appropriate by FDA. The accuracy, defined as positive percent agreement (PPA) and negative percent agreement (NPA), must be measured; accuracy point estimates must be > 99% (both per reported variant and overall) and uncertainty of the point estimate must be presented using the 95% confidence interval. Clinical specimens must include both homozygous wild type and heterozygous genotypes. The number of clinical specimens for each variant reported that must be included in the accuracy study must be based on the variant prevalence. Common variants (> 0.1% allele frequency in ethnically relevant population) must have at least 20 unique heterozvgous clinical specimens tested. Rare variants (≤ 0.1% allele frequency in ethnically relevant population) shall have at least 3 unique mutant heterozygous specimens tested.

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. Variants that have a point estimate for PPA or NPA of