The GlutenID Celiac Genetic Health Risk Test uses qualitative genotyping to detect clinically relevant variants in genomic DNA isolated from salva collected from individuals 18 years of age or older with ORAcollect Dx OCD-100 for the purpose of reporting and interpreting Genetic Health Risks (GHR). The GlutenID Celiac GHR Test is indicated for reporting of one variant associated with the HLA-DQ2.5 haplotype, one variant associated with the HLA-DQ8 haplotype, one variant associated with the HLA-DQ7 haplotype, and three variants associated with the HLA-DQ2.2 haplotype. The report describes if a person has variants linked to haplotypes associated with an increased risk for developing celiac disease, but it does not describe a person's overall risk of developing celiac disease. This report is most relevant for people of European descent.
Device Story
The GlutenID Celiac Genetic Health Risk Test is an in vitro diagnostic device that performs qualitative genotyping on genomic DNA. Input consists of saliva samples collected using the ORAcollect Dx OCD-100 collection kit. The device analyzes DNA to identify specific genetic variants associated with HLA-DQ2.5, HLA-DQ8, HLA-DQ7, and HLA-DQ2.2 haplotypes. The output is a report indicating the presence of these variants, which are linked to an increased risk of developing celiac disease. The test is intended for use by adults 18 years or older. The report provides information regarding genetic risk factors but does not determine an individual's overall risk of developing celiac disease. Healthcare providers use this information to assess genetic predisposition in patients, particularly those of European descent, to inform clinical discussions regarding celiac disease risk.
Clinical Evidence
Bench testing only. Analytical performance validated via precision/reproducibility studies (n=270), DNA extraction precision (n=144), lot-to-lot precision (n=54), and interference studies (endogenous, exogenous, smoking, microbial). Accuracy established by comparing 143 clinical samples against bi-directional Sanger sequencing, showing 100% concordance. User comprehension study (n=314) confirmed participants understood test purpose, limitations, and results with >94% post-test comprehension scores.
Technological Characteristics
Qualitative genotyping assay for detection of specific genomic DNA variants. Utilizes saliva samples collected via ORAcollect Dx OCD-100. Analyzes HLA-DQ2.5, HLA-DQ8, HLA-DQ7, and HLA-DQ2.2 haplotype-associated variants.
Indications for Use
Indicated for individuals ≥ 18 years old to detect clinically relevant variants in genomic DNA from saliva associated with HLA-DQ2.5, HLA-DQ8, HLA-DQ7, and HLA-DQ2.2 haplotypes to report genetic health risk for celiac disease. Not for diagnosis; not a substitute for genetic counseling or medical consultation.
Regulatory Classification
Identification
A genetic health risk assessment system is a qualitative in vitro molecular diagnostic system used for detecting variants in genomic deoxyribonucleic acid (DNA) isolated from human specimens that will provide information to users about their genetic risk of developing a disease to inform lifestyle choices and/or conversations with a health care professional. This assessment system is for over-the-counter use. This device does not determine the person's overall risk of developing a disease.
Special Controls
*Classification.* Class II (special controls). The genetic health risk assessment system device, when it has previously received a first-time FDA marketing authorization (*e.g.,* 510(k) clearance) for the genetic health risk assessment system (a “one-time FDA reviewed genetic health risk assessment system”), is exempt from the premarket notification procedures in part 807, subpart E, of this chapter subject to the limitations in § 866.9. The device must comply with the following special controls:(1) The 21 CFR 809.10 compliant labeling and any prepurchase page and test report generated, unless otherwise specified, must include:
(i) A section addressed to users with the following information:
(A) The limiting statement explaining that this test provides genetic risk information based on assessment of specific genetic variants but does not report on a user's entire genetic profile. This test [does not/may not, as appropriate] detect all genetic variants related to a given disease, and the absence of a variant tested does not rule out the presence of other genetic variants that may be related to the disease.
(B) The 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.
(C) The limiting statement explaining that other factors such as environmental and lifestyle risk factors may affect the risk of developing a given disease.
(D) The 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 health care professional prior to collection of a sample for testing. This test is not a substitute for visits to a doctor or other health care professional. Users should consult with their doctor or other health care professional if they have any questions or concerns about the results of their test or their current state of health.
(E) Information about how to obtain access to a genetic counselor, board-certified clinical molecular geneticist, or equivalent health care professional about the results of a user's test.
(F) The limiting statement explaining that this test is not intended to diagnose a disease, 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.
(G) 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.
(ii) A section in your 21 CFR 809.10 labeling and any test report generated that is for health care professionals who may receive the test results from their patients with the following information:
(A) The limiting statement explaining that this test is not intended to diagnose a disease, determine medical treatment, or tell the user anything about their current state of health.
(B) The limiting statement explaining that this test is intended to provide users with their genetic information to inform lifestyle decisions and conversations with their doctor or other health care professional.
(C) The limiting statement explaining that any diagnostic or treatment decisions should be based on testing and/or other information that you determine to be appropriate for your patient.
(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 Web site where the manufacturer shall 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 Web site 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 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 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) An index of the material being provided to meet the requirements in paragraph (b)(3) of this section and its location.
(ii) 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. If there are different categories of risk, the manufacturer must provide literature references 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, but is not limited to, 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* )*Prepurchase page:* This page must contain information that informs the user about what information the test will provide. This includes, but is not limited to, variant information, the condition or disease 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) 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 section must be provided. This opt-in page must be provided for each disease 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* ) Professional guidelines that recommend when genetic testing for the associated target condition is or is not recommended; and(
*iv* ) A recommendation to speak with a health care professional, genetic counselor, or equivalent professional before getting the results of the test.(
*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 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 followup procedures, how the results of the test may affect the user's family, including children, and links to resources that provide additional information.(iii) A technical information section containing the following information:
(A) Gene(s) and variant(s) the test detects using standardized nomenclature, Human Genome Organization nomenclature and coordinates as well as Single Nucleotide Polymorphism Database (dbSNP) reference SNP numbers (rs#).
(B) Scientifically established disease-risk association of each variant detected and reported by the test. This risk association information must include:
(
*1* ) Genotype-phenotype information for the reported variants.(
*2* ) Table of expected frequency and risks of developing the disease in relevant ethnic populations and the general population.(
*3* ) A statement about the current professional guidelines for testing these specific gene(s) and variant(s).(
*i* ) If professional guidelines are available, provide the recommendations in the professional guideline 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.(
*ii* ) If professional guidelines are not available, provide a statement that the professional guidelines are not available for these specific gene(s) and variant(s).(C) The specimen type (
*e.g.,* saliva, capillary whole blood).(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 (
*i.e.,* percentage of tests that failed quality control) based on data from clinical samples, a description of scenarios in which a test can fail (*i.e.,* low sample volume, low DNA concentration, etc.), how users will be notified of a test failure, and the nature of followup actions on a failed test to be taken by the user and the manufacturer.(I) Specification of the criteria for test result interpretation and reporting.
(J) Information that demonstrates the performance characteristics of the test, including:
(
*1* ) Accuracy of study results for each claimed specimen type.(
*i* ) Accuracy of the test shall 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 shall 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 shall mimic clinical specimens as much as is feasible and provide an unbiased evaluation of the device accuracy.(
*ii* ) 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 device must be predefined and appropriate to the device's intended use. Detailed study protocols must be provided.(
*iii* ) Test specimens 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 using either the minimum numbers of samples identified in this paragraph or, when determined appropriate and identified by FDA, a minimum number of samples determined using an alternative method. When appropriate, the same samples may be used in testing to demonstrate the accuracy of testing for multiple genotypes by generating sequence information at multiple relevant genetic locations. At least 20 unique samples representing the wild-type genotype must be tested. To test samples that are heterozygous for the reported variant(s), common variants (>0.1 percent variant frequency in the relevant population) must be tested with at least 20 unique samples. Rare variants (≤0.1 percent variant frequency in the relevant population) must be tested with at least three unique samples. To test samples that are homozygous for the reported variant(s), variants with ≥2 percent variant frequency in a relevant population must be tested with at least 20 unique samples. Variants with a frequency in the relevant population <2 percent and ≥0.5 percent must be tested with at least 10 unique samples. Variants with a frequency in the relevant population <0.5 percent must be tested with at least three unique samples. If variants with a frequency of <0.5 percent are not found within the relevant population and homozygous samples are not tested, then the test results for this homozygous rare variant must not be reported to the user.(
*iv* ) Information about the accuracy study shall include the number and type of samples that were compared to bidirectional Sanger sequencing or other methods identified as appropriate by FDA. This information must either be reported in tabular format and arranged by clinically relevant variants or reported using another method identified as appropriate by FDA. As an example, for samples with different genotypes DD, Dd, and dd, the following table represents data from the accuracy study presented in tabular format: (
*v* ) The accuracy represents the degrees of agreement between the device results and the comparator results. The accuracy must be evaluated by measuring different percent agreements (PA) of device results with the comparator results and percent of 'no calls' or 'invalid calls.' Calculate the rate of 'no calls' and 'invalid calls' for each comparator output as %Inv(DD) = A4 /NDD, %Inv(Dd) = B4 /NDd , %Inv(dd) = C4 /Ndd . If 'no calls' or 'invalid calls' are required to be retested according to the device instructions for use, the percent of final 'no calls' or 'invalid calls' must be provided. In the table presenting the results of the accuracy study, use only the final results (*i.e.,* after retesting the initial 'no calls' or 'invalid calls', if required according to the instructions for use). Samples that resulted in a 'no call' or 'invalid call' after retesting must not be included in the final calculations of agreement. If the percentages of 'no calls' or 'invalid calls' for each comparator output are similar, combine these estimates as (A4 + B4 + C4 )/(NDD + NDd + Ndd ) and provide a 95 percent two-sided confidence interval. The percent of final 'no calls' or 'invalid calls' must be clinically acceptable.(
*vi* ) Point estimates of percent agreement 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 excluding 'no calls' or 'invalid calls'. The calculations must be performed as follows: (
*vii* ) For percent agreements for DD, Dd and dd (PA(DD|DD), PA(Dd|Dd) and PA(dd|dd)) as described in paragraph (b)(3)(iii)(J)(*1* )(*vi* ) of this section, the 95 percent two-sided confidence intervals must be provided. The accuracy point estimates for percent agreements for DD, Dd and dd must be ≥99 percent per reported variant and overall. Any variants that have a point estimate for either PA(DD|DD), PA(Dd|Dd), or PA(dd|dd) of <99 percent compared to bidirectional sequencing or other methods identified as appropriate by FDA must not be incorporated into test claims and reports. Accuracy results 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 type and by genotype or must be reported using another method identified as appropriate by FDA (see paragraph (b)(3)(iii)(J)(*1* )(*iv* ) of this section).(
*viii* ) Information must be reported on the Technical Positive Predictive Value (TPPV) related to the analytical (technical) performance of the device for genotypes in each relevant subpopulation (*e.g.,* ethnicity, gender, age, geographical location, etc.). TPPV is the percentage of individuals with the genotype truly present among individuals whose test reports indicate that this genotype is present. The TPPV depends on the accuracy measures of percent agreements and on the frequency of the genotypes in the subpopulation being studied. The f(DD) is the frequency of DD and f(Dd) is the frequency of Dd in the subpopulation being studied; TPPV must be calculated as described in paragraphs (b)(3)(iii)(J)(*1* )(*ix* ) through (*xi* ) of this section.(
*ix* ) For variants where the point estimates of PA(DD|DD), PA(Dd|Dd) and PA(dd|dd) are less than 100 percent, use these point estimates in TPPV calculations.(
*x* ) Point estimates of 100 percent in the accuracy study may have high uncertainty about performance of the test in the population. If these variants are measured using highly multiplexed technology, calculate the random error rate for the overall device. The accuracy study described in paragraph (b)(3)(iii)(J) of this section in those cases is more to determine that there is no systematic error in such devices. In those cases, incorporate that rate in the estimation of the percent agreements as calculated in paragraph (b)(3)(iii)(J)(*1* )(*vi* ) of this section and include it in TPPV calculations.(
*xi* ) The TPPV for subpopulations with genotype frequencies of f(dd), f(Dd) and f(DD) = 1−f(dd)−f(Dd) in the subpopulation is calculated as: (
*2* ) 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 either include specimens from the claimed sample type (*e.g.,* saliva) representing all genotypes for each variant (*e.g.,* wild type, heterozygous, and homozygous) or, if an alternative panel composition of specimens is identified by FDA as appropriate, a panel composed of those specimens FDA identified as appropriate. A detailed study protocol must be created in advance of the study and must include predetermined acceptance criteria for performance results. The percentage of samples that failed quality control 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 shall report the variants tested in the study and the number of replicates for each variant, and what conditions were tested (*i.e.,* number of runs, days, instruments, reagent lots, operators, specimens/type, etc.). Results must be evaluated and presented in tabular format and stratified by study parameter (*e.g.,* by site, instrument(s), reagent lot, operator, and sample variant). The study must include all extraction steps from the claimed specimen type or matrix, unless a separate extraction reproducibility 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 reproducibility 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.(
*3* )*Analytical specificity data:* Data must be provided that evaluates the effect of potential endogenous and exogenous interferents on test performance, including specimen extraction and variant detection. Interferents tested must include those reasonably likely to be potentially relevant to the sample type used for the device.(
*4* )*Interfering variant data:* Nucleotide mutations that can interfere with the technology must be cited and evaluated. Data must be provided to demonstrate the effect of the interfering variant(s) on the performance of the correct calls. Alternatively, for each suspected interfering mutation for which data is not provided demonstrating the effect of the interfering variant, the manufacturer must identify the suspected interfering variants in the labeling and indicate that the impact that the interfering variants may have on the assay's performance has not been studied by providing a statement that reads “It is possible that the presence of [insert clearly 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.”(
*5* )*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.(
*6* )*Reagent stability:* The manufacturer must evaluate reagent stability using wild-type, heterozygous, and homozygous samples. Reagent stability data must demonstrate that the reagents maintain the claimed accuracy and reproducibility. Data supporting such claims must be provided.(
*7* )*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.(K) Clinical performance summary.
(
*1* ) Information to support the clinical performance of each variant reported by the test must be provided.(
*2* ) 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.(
*i* ) If odds ratios are available, using information about the genotype distribution either among individuals with the target disease absent, or in the general population, or information about the risk variant frequency and odds ratios, the likelihood ratios for the corresponding device results along with 95 percent confidence intervals must be calculated. Using information about pretest risk (π), an estimate of likelihood ratio (LR), and a relationship between post-test risk R as R/(1−R) = LR·π/(1−π), the post-test risk R must be calculated.(
*ii* ) When available, likelihood ratios (LR) for different test results must be presented in a tabular format along with references to the source data or using another method identified as appropriate by FDA as stated in paragraph (b)(3)(iii)(K)(2) of this section. When these values are not directly available in published literature, likelihood ratios can be separately calculated along with the 95 percent confidence interval with references to the source data. Note that a minimum requirement for the presence of the variant's effect on the risk is that a corresponding LR is statistically higher than 1 (a lower bound of 95 percent two-sided confidence interval is larger than 1). It means that the post-test risk is statistically higher than the pretest risk (an observed value of the difference between the post-test and pretest risks).(L) Materials that explain the main concepts and terminology used in the test that includes, but is not limited to:
(
*1* )*Definitions:* Scientific terms that are used in the test reports.(
*2* )*Prepurchase page:* This page must contain information that informs the user about what the test will provide. This includes, but is not limited to, variant information, the condition or disease 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) 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 section must be provided. This opt-in page must be provided for each disease 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* ) Professional guidelines that recommend when genetic testing for the associated target condition is or is not recommended; and(
*iv* ) A recommendation to speak with a health care professional, genetic counselor, or equivalent professional before getting the results of the test.(
*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 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 on the test results, information about the population to which the variants in the test is most applicable, the meaning of the result(s), other risks factors that contribute to disease, appropriate followup procedures, how the results of the test may affect the user's family, including children, and links to resources that provide additional information.(M) 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.
(
*1* ) The test manufacturer must provide a genetic 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.(
*2* ) The test manufacturer must perform pre- and post-test user comprehension studies. The comprehension test questions must include directly evaluating a representative sample of the material being presented to the user as described in paragraph (b)(3)(ii) of this section.(
*3* ) 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.(
*4* ) The user study must meet the following criteria:(
*i* ) 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 shall be well defined in the inclusion and exclusion criteria.(
*ii* ) All sources of bias must be predefined and accounted for in the study results with regard to both responders and non-responders.(
*iii* ) 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).(
*iv* ) 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 of 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.(
*v* ) 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.(
*vi* ) 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.(
*vii* ) The analysis of the user comprehension results must include results regarding reports that are provided for each gene/variant/ethnicity tested, statistical methods used to analyze all data sets, and 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, etc.) for each study report must be included.(4) The intended use of the device must not include the following indications for use:
(i) Prenatal testing;
(ii) Determining predisposition for cancer where the result of the test may lead to prophylactic screening, confirmatory procedures, or treatments that may incur morbidity or mortality to the patient;
(iii) Assessing the presence of genetic variants that impact the metabolism, exposure, response, risk of adverse events, dosing, or mechanisms of prescription or over-the-counter medications; or
(iv) Assessing the presence of deterministic autosomal dominant variants.
Related Devices
K123713 — IMMULISA ENHANCED CELIAC FUSION (TTG/DGP) IGA/IGG ANTIBODY ELISA · Immco Diagnostics, Inc. · Oct 25, 2013
K091522 — IMMULISA CELIAC G+ (GLIADIN) IGA AND IGG ANTIBODY ELISA · Immco Diagnostics, Inc. · Feb 4, 2010
K052143 — QUANTA LITE GLIADIN IGA II · Inova Diagnostics, Inc. · Oct 4, 2005
K062708 — QUANTA LITE CELIAC DGP SCREEN · Inova Diagnostics, Inc. · Dec 13, 2006
Submission Summary (Full Text)
{0}
FDA
U.S. FOOD & DRUG
ADMINISTRATION
# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
ASSAY ONLY
## I Background Information:
A 510(k) Number
K241456
B Applicant
Targeted Genomics, LLC
C Proprietary and Established Names
GlutenID Celiac Genetic Health Risk Test
D Regulatory Information
| Product Code(s) | Classification | Regulation Section | Panel |
| --- | --- | --- | --- |
| PTA | Class II | 21 CFR 866.5950 - Genetic Health Risk Assessment System | IM - Immunology |
## II Submission/Device Overview:
A Purpose for Submission:
New device
B Measurand:
Variants associated with HLA-DQ2.5, HLA-DQ8, HLA-DQ7 and HLA-DQ2.2 haplotypes, in genomic DNA obtained from a human saliva sample.
C Type of Test:
Over-the-counter (OTC) genetic testing service, qualitative single nucleotide polymorphism (SNP) genotyping using next generation sequencing (NGS)
## III Intended Use/Indications for Use:
Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
{1}
K241456 - Page 2 of 24
A Intended Use(s):
See Indications for Use below.
B Indication(s) for Use:
The GlutenID Celiac Genetic Health Risk Test uses qualitative genotyping to detect clinically relevant variants in genomic DNA isolated from saliva collected from individuals 18 years of age or older with ORAcollect Dx OCD-100 for the purpose of reporting and interpreting Genetic Health Risks (GHR).
The GlutenID Celiac GHR Test is indicated for reporting of one variant associated with the HLA-DQ2.5 haplotype, one variant associated with the HLA-DQ8 haplotype, one variant associated with the HLA-DQ7 haplotype, and three variants associated with the HLA-DQ2.2 haplotype. The report describes if a person has variants linked to haplotypes associated with an increased risk for developing celiac disease, but it does not describe a person's overall risk of developing celiac disease. This report is most relevant for people of European descent.
C Special Conditions for Use Statement(s):
- For over-the-counter (OTC) use.
- The test is intended for users ≥ 18 years old.
- This test is not a substitute for visits to a genetic counselor or healthcare provider. It is recommended that the user consults with a genetic counselor or healthcare provider if there are any questions or concerns about the results.
- The test does not detect all genetic variants related to celiac disease. The absence of a variant tested does not rule out the presence of other genetic variants that may be celiac disease related.
- The test does not diagnose any specific health conditions, determine medical treatment or other medical intervention, or tell the user anything about their current state of health. Only a healthcare provider can diagnose celiac disease. Results from this test should not be used solely to make medical decisions.
- If the user has other risk factors for the celiac disease, they should discuss this with a genetic counselor or healthcare provider.
- A user's race, ethnicity, age, sex, and lifestyle may affect how the genetic results are interpreted.
- The laboratory may not be able to process a patient's sample due to low DNA quantity/quality. This is estimated to occur with a frequency of less than 1%. If this happens, the user will receive an email notification.
- Subject to meeting the limitations contained in the special controls under regulation 21 CFR 866.5950.
D Special Instrument Requirements:
{2}
The GlutenID Celiac GHR Test is to be performed on the Illumina MiSeqDx instrument (DEN130011).
The GlutenID Celiac GHR Test software is comprised of two modules. The Pipeline module serves to process sequencing data from the Illumina MiSeqDx instrument into annotated variants. The Reporting module generates final reports for individuals based on the annotated variants results.
## IV Device/System Characteristics:
## A Device Description:
The GlutenID Celiac GHR Test is a direct-to-consumer DNA genetic test intended for OTC use. The test uses qualitative genotyping to detect clinically relevant genetic variants associated with the risk of developing celiac disease. The GlutenID Celiac GHR Test is indicated for reporting of one variant associated with the HLA-DQ2.5 haplotype, one variant associated with the HLA-DQ8 haplotype, one variant associated with the HLA-DQ7 haplotype, and three variants associated with the HLA-DQ2.2 haplotype as shown below:
| Haplotype | SNP ID |
| --- | --- |
| HLA-DQ 2.5* | rs2187668 |
| HLA-DQ8 | rs7454108 |
| HLA-DQ7 | rs4639334 |
| HLA-DQ2.2^ | rs2395182 |
| | rs7775228 |
| | rs4713586 |
* DQ2.5 haplotype is commonly referred to as DQ2(cis)
^ DQ2.2 haplotype can be referred to as half-DQ2
The tests are conducted with genomic DNA isolated from human saliva sample collected in the ORAcollect Dx OCD-100 (cleared under K212745).
The GlutenID Celiac GHR Test is composed of:
(i) ORAcollect-Dx OCD-100 is intended for use in the collection of saliva samples.
(ii) GlutenID Celiac GHR Test is intended for the genetic analysis and detection of genetic variants associated with the HLA-DQ haplotypes using NGS.
(iii) GlutenID Pipeline Software is developed and validated on the Galaxy Platform as a modular workflow and data analysis application used to view and analyze genotyping data from the Illumina MiSeqDx. The GID Pipeline outputs base calling, read alignment, and variant identification.
(iv) The GlutenID Reporting Software is developed and validated to automate variant annotation from the technical data. It provides a Consolidated Report with predicted GlutenID genotype for each sample. Individual GlutenID Final Reports are automatically generated based on the predicted GlutenID from the Consolidated Report.
K241456 - Page 3 of 24
{3}
(v) GlutenID Celiac GHR website and result portal software is intended to provide the contents and the procedure to order and use the OTC Service.
A user's saliva is self-collected using ORAcollect Dx OCD-100 device which consists of a collection tube containing a stabilizing buffer solution. The collected sample is shipped to a Clinical Laboratory Improvement Amendments (CLIA) certified laboratory for processing, testing and analysis. Human genomic DNA isolated from the saliva sample is isolated, processed and sequenced using NGS reagents on the Illumina MiSeqDx instrument. Following completion of a sequencing run, the data is analyzed using MiSeq integrated system software and genotyping results of the six celiac genetic risk variants are automatically processed into one of 15 associated GlutenIDs genotypes. The personalized final reports for each sample are generated using the GlutenID Reporting Software to provide information about the celiac disease genetic risk associated with their assigned genotypes.
The GlutenID Risk category for each of the 15 GlutenID genotypes are shown in the following table:
| GlutenID Genotype | Estimated Risk | Risk Category |
| --- | --- | --- |
| DQ2+DQ2 | 1 in 9 (8%) | Increased |
| DQ2+half-DQ2 | 1 in 9 (8%) | |
| DQ2+DQ8 | 1 in 12 (6%) | |
| DQ8+half-DQ2 | 1 in 25 (4%) | |
| DQ2+DQ7 | 1 in 35 (3%) | |
| DQ2.2+DQ7 [DQ2 (trans)] | 1 in 35 (3%) | |
| DQ2 (cis) | 1 in 35 (3%) | |
| DQ8+DQ8 | 1 in 100 (1%) | Low |
| DQ8 | 1 in 100 (1%) | |
| DQ8+DQ7 | 1 in 100 (1%) | |
| Half-DQ2+half-DQ2 (DQ2.2+DQ2.2) | 1 in 210 (0.5%) | Not likely at risk |
| Half-DQ2 (DQ2.2) | 1 in 210 (0.5%) | |
| DQ7+DQ7 | 1 in 1842 (0.05%) | |
| DQ7 | 1 in 1842 (0.05%) | |
| Non-celiac genetics (NCG) | 1 in 2518 (0.05%) | |
Note: DQ2(cis) refers to DQ2.5 haplotype, half-DQ2 refers to DQ2.2 haplotype, DQ7+DQ2.2 refers to DQ2(trans).
## B Principle of Operation:
The GlutenID Celiac GHR Test is performed by a CLIA-certified laboratory. The test is a qualitative single nucleotide polymorphism (SNP) genotyping assay that is performed for each sample using NGS to detect the variants associated with the haplotypes HLA-DQ2.5, HLA-DQ8, HLA-DQ7, HLA-DQ2.2.
Genomic DNA is extracted from human saliva samples collected using ORAcollect·Dx OCD-100. Celiac disease associated SNP genomic regions of each DNA sample are amplified. This results in the generation of sample libraries that are transferred into a MiSeqDx reagent cartridge containing all required reagents. The cartridge is inserted into the MiSeqDx instrument which
K241456 - Page 4 of 24
{4}
performs cluster generation, sequencing, and data analysis. The raw data are uploaded onto the Galaxy Platform and analyzed using the GlutenID Pipeline Software. Genotyping results of the six celiac disease genetic risk variants are automatically processed into one of 15 associated GlutenID genotypes by the GlutenID Reporting Software. The GlutenID Celiac GHR Test interprets and categorizes each user's Gluten ID genotype into risk categories (Not likely at risk, Low risk, and Increased risk) based on the estimated lifetime risk of celiac disease.
V Substantial Equivalence Information:
A Predicate Device Name(s):
23andMe Personal Genome Service (PGS) Genetic Health Risk Test for Celiac Disease
B Predicate 510(k) Number(s):
DEN160026
C Comparison with Predicate(s):
| Device & Predicate Device(s): | K241456 | DEN160026 (Predicate) |
| --- | --- | --- |
| Device Trade Name | GlutenID Celiac Genetic Health Risk (GHR) Test | 23andMe Personal Genome Service (PGS) Genetic Health Risk Test for Celiac Disease |
| General Device Characteristic Similarities | | |
| Intended Use/Indications For Use | The GlutenID Celiac Genetic Health Risk Test uses qualitative genotyping to detect clinically relevant variants in genomic DNA isolated from saliva collected from individuals 18 years of age or older with ORAcollect Dx OCD-100 for the purpose of reporting and interpreting Genetic Health Risks (GHR).The GlutenID Celiac GHR Test is indicated for reporting of one variant associated with the HLA-DQ2.5 haplotype, one variant associated with the HLA-DQ8 haplotype, one variant associated with the HLA-DQ7 haplotype, and three variants associated with the HLA-DQ2.2 haplotype. The report describes if a person has variants linked to haplotypes associated with an | The 23andMe Personal Genome Service (PGS) Test uses qualitative genotyping to detect the following clinically relevant variants in genomic DNA isolated from human saliva collected from individuals ≥18 years with the Oragene Dx model OGD-500.001 for the purpose of reporting and interpreting Genetic Health Risks (GHR).The 23andMe PGS Genetic Health Risk Report for Celiac Disease is indicated for reporting of a variant associated with the HLA-DQ2.5 haplotype. The report describes if a person has a haplotype associated with an increased risk of developing celiac disease, but it does not describe a person's overall risk for |
K241456 - Page 5 of 24
{5}
K241456 - Page 6 of 24
| | increased risk for developing celiac disease, but it does not describe a person’s overall risk of developing celiac disease. This report is most relevant for people of European descent. | developing celiac disease. This report is most relevant for people of European descent. |
| --- | --- | --- |
| Special Conditions for Use | • For over-the-counter (OTC) use.
• The test is intended for users ≥ 18 years old.
• This test is not a substitute for visits to a genetic counselor or healthcare provider. It is recommended that the user consults with a genetic counselor or healthcare provider if there are any questions or concerns about the results.
• The test does not detect all genetic variants related to celiac disease. The absence of a variant tested does not rule out the presence of other genetic variants that may be celiac disease related.
• The test does not diagnose any specific health conditions, determine medical treatment or other medical intervention, or tell the user anything about their current state of health. Only a healthcare provider can diagnose celiac disease. Results from this test should not be used solely to make medical decisions.
• If the user has other risk factors for the celiac disease, they should discuss this with a genetic counselor or healthcare provider.
• A user's race, ethnicity, age, sex, and lifestyle may affect how the genetic results are interpreted.
• The laboratory may not be able to process a patient’s sample due to low DNA quantity/quality. This is estimated to occur with a frequency of less than 1%. If this happens, the user will receive an email notification.
• Subject to meeting the limitations contained in the special controls under regulation 21 CFR 866.5950. | • For over-the-counter (OTC) use.
• 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.
• The 23andMe PGS Genetic Health Risk Tests for Celiac disease, do not detect all genetic variants associated with the celiac disease. 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 for users ≥ 18 years old.
• The test does not diagnose any specific health conditions. Results should not be used to make medical decisions.
• 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 7.6%.
• A user’s race, ethnicity, age, and sex may affect how the genetic test results are interpreted.
• Subject to meeting the limitations contained in the special controls under regulation 21 CFR 866.5950. |
{6}
| Type of Test | Qualitative genotyping for allelic variant determination through SNP detection | Same |
| --- | --- | --- |
| Design | Software application that includes product information page, e-commerce (registration and order DNA kit), secure login, download genetic report | Same |
| Interpretation of Results | For OTC use. Specialized interpretation by a physician not required | Same |
| Specimen Type | Saliva | Same |
| Measurand | Genomic DNA | Same |
| Human Factors | User Comprehension Study | Same |
| Intended User | ≥ 18 years old | Same |
| General Device Characteristic Differences | | |
| Specimen Collection | ORAcollect Dx model OCD-100 | Oragene Dx model OGD-500.001 |
| Variants Detected | HLA-DQA1, HLA-DQB1 haplotypes DQ2.5, DQ8, DQ7, DQ2.2 | HLA-DQA1, HLA-DQB1 haplotypes DQ2.5 |
| Technology | Next generation sequencing (NGS) | Microarray genotyping |
| Instrument | Illumina MiSeqDx System | Tecan Evo and Illumina iScan |
| Software | The GlutenID Pipeline Software application is a modular software used to view and analyze genomic data from the MiSeqDx. The GlutenID Reporting software application conducts a variety of control checks on the file resulting in automated technical reporting of the sample's genotype profile. Final reports with associated disease risk are automatically generated from the technical report data. | GenomeStudio is a modular software application that is used to view and analyze genotype data obtained from the iScan. Coregen software conducts a variety of control checks on the file, resulting in a final genotype profile for each sample These data are used to generate test reports on a user's genotype and associated risk of disease |
VI Standards/Guidance Documents Referenced:
- CLSI EP12-Ed3, Evaluation of Qualitative, Binary Output Examination Performance
- Class II Special Controls: 866.5950 - Code of Federal Regulations Title 21
- Guidance for the Content of Premarket Submissions for Software Contained in Medical Devices (2005)
- General Principles of Software Validation Guidance for Industry and FDA Staff (2002)
- Off-The-Shelf Software Use in Medical Devices Guidance for Industry and Food and Drug Administration Staff (2019)
K241456 - Page 7 of 24
{7}
- Content of Premarket Submissions for Management of Cybersecurity in Medical Devices (2014)
## VII Performance Characteristics (if/when applicable):
## A Analytical Performance:
The test performance was validated at the single site, and the results presented below met the pre-defined acceptance criteria outlined in the Special Controls of 21 CFR 866.5950. Information regarding samples that failed quality control (FQC) was also evaluated and presented in each study below.
## 1. Precision/Reproducibility:
An evaluation of precision of the GlutenID Celiac GHR Test was performed by testing 30 samples representing variants for each celiac genetic risk haplotypes: DQ2.2, n=5; DQ2.5, n=7; DQ7, n=4; DQ8, n=7; DQ2+DQ2.2, n=2; DQ2+DQ8, n=1; DQ2.2+DQ7 [DQ2(trans)], n=4. Testing included using three reagent lots, three MiSeqDx analyzers, two thermocyclers and three different operators. Each sample was run in three replicates per run, three runs per day for three non-consecutive days, for a total of 270 results. Each run combined different reagent lots, instruments, and operators at a single site. Relevant genotypes for the samples were confirmed by bi-directional Sanger sequencing. Number of correct calls, number of failed QC (FQC), % of FQC, and %correct calls were analyzed. The results are presented in the following table:
| Haplotype | No. of Samples | No. of Replicates | No. of Correct Calls | No. of FQC | % of FQCs | % of Correct Calls |
| --- | --- | --- | --- | --- | --- | --- |
| DQ2.2 | 5 | 45 | 45 | 0 | 0% | 100% |
| DQ2.5 | 7 | 63 | 63 | 0 | 0% | 100% |
| DQ7 | 4 | 36 | 36 | 0 | 0% | 100% |
| DQ8 | 7 | 63 | 63 | 0 | 0% | 100% |
| DQ2+DQ2.2 | 2 | 18 | 18 | 0 | 0% | 100% |
| DQ2.2+DQ7
DQ2(trans) | 4 | 36 | 36 | 0 | 0% | 100% |
| DQ2+DQ8 | 1 | 9 | 9 | 0 | 0% | 100% |
| Total | 30 | 270 | 270 | 0 | 0% | 100% |
- No samples failed QC (FQC); percentage of FQC was 0% (0/270) with 95% confidence interval (CI): (0.0%; 1.4%).
- A total of 270 results were in the precision study across three lots, three operators, three days, and three machine combinations. Percentage of correct calls was 100% (270/270) with 95% CI: (98.6%; 100%).
## 2. DNA Extraction Precision Study
To evaluate the effect of the extraction step on samples used with GlutenID Celiac GHR Test, a separate DNA extraction study was conducted. Three saliva samples were self-
K241456 - Page 8 of 24
{8}
collected from each of 12 donors representing the four haplotypes (DQ2.5, n=6; DQ8, n=2; DQ7, n=2; DQ2.2, n=2). Two operators performed one independent DNA extraction on each of three non-consecutive days for a total of six runs of DNA extraction per subject. Each extracted DNA was tested with the GlutenID Celiac GHR Test in duplicate. A total of two GlutenID Celiac GHR Test runs were performed (Run 1 with the DNA extractions from operator 1 and Run 2 with the DNA extractions from operator 2) and the total number of results per subject was 12 (3 samples x 4 results/sample). The total number of DNA extractions was 72 with the total number of 144 over the course of the study (12 subjects x 2 operators x 3 days x 1 DNA extraction per operator per sample per day x 2 replicates). The results are summarized in the following table:
| Haplotype | No. of Subjects | No. of Samples | No. of Extractions | No. of Replicates | No. of Correct Calls | No. of FQCs | % of FQCs | % of Correct Calls |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| DQ2.2 | 2 | 6 | 12 | 24 | 24 | 0 | 0% | 100% |
| DQ2.5 | 6 | 18 | 36 | 72 | 72 | 0 | 0% | 100% |
| DQ7 | 2 | 6 | 12 | 24 | 24 | 0 | 0% | 100% |
| DQ8 | 2 | 6 | 12 | 24 | 24 | 0 | 0% | 100% |
| Total | 12 | 36 | 72 | 144 | 144 | 0 | 0% | 100% |
- No samples failed QC (FQC); %FQC was 0% (0/144) with 95% CI: (0.0%; 2.6%).
- Percentage of correct calls was 100% (144/144) with 95% CI: (97.4%; 100%)
## 3. Lot-to-Lot Precision
To evaluate the lot-to-lot imprecision for the GlutenID Celiac GHR Test, a study was conducted by three operators who prepared three different reagent lots to ensure three lots contained different manufacturing preparation of the critical reagents. Each operator tested a panel of six saliva DNA samples representing the DQ2, DQ2+DQ2, DQ8, DQ8+DQ8, and DQ2.2+DQ7 genotypes in triplicate using one of the three lots, resulting a total of 18 results (6 sample x 3 replicates) per lot and a total of 54 replicates (18 replicates x 3 lots). The results are summarized in the following table:
| Haplotype | No. of Replicates | No. of Correct Calls | | | No. of FQCs | | | % of FQCs | % of Correct Calls |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | Lot 1 | Lot 2 | Lot 3 | Lot 1 | Lot 2 | Lot 3 | | |
| DQ2 | 9 | 3 | 3 | 3 | 0 | 0 | 0 | 0% | 100% |
| DQ2+DQ2 | 9 | 3 | 3 | 3 | 0 | 0 | 0 | 0% | 100% |
| DQ8 | 9 | 3 | 3 | 3 | 0 | 0 | 0 | 0% | 100% |
| DQ8+DQ8 | 9 | 3 | 3 | 3 | 0 | 0 | 0 | 0% | 100% |
| DQ2.2+DQ7 | 9 | 3 | 3 | 3 | 0 | 0 | 0 | 0% | 100% |
| DQ2.2+DQ7 | 9 | 3 | 3 | 3 | 0 | 0 | 0 | 0% | 100% |
| Total | 54 | 18 | 18 | 18 | 0 | 0 | 0 | 0% | 100% |
- No samples failed QC (FQC); percentage of FQC was 0% (0/54) with 95% CI: (0.0%; 6.6%).
- Percentage of correct calls 100% (54/54) across three reagent lots with 95% CI: (93.4%; 100%)
K241456 - Page 9 of 24
{9}
4. Linearity:
Not applicable
5. Analytical Specificity/Interference:
Saliva samples collected using the ORAcollect Dx OCD-100 device were evaluated for the impact of endogenous, exogenous, smoking, and microbial substances, on the performance of the GlutenID Celiac GHR Test.
i. Endogenous Interference
An endogenous interference study was conducted to determine whether potential endogenous substances present during saliva collection affect the performance of the GlutenID Celiac GHR Test. Saliva samples were self-collected from five individuals (representing the four haplotypes DQ2.2, DQ2.5, DQ7, and DQ8). Samples were split and individual aliquots were spiked with the following:
- salivary amylase: 395 U/ml
- hemoglobin: 20 mg/mL
- IgA: 0.43 mg/mL
- Total protein: 2.67 mg/mL (composed of 0.185 mg/mL salivary amylase, 0.43 mg/mL IgA, and 2.05 mg/ml Human Serum Albumin)
Three replicates were tested for each aliquot. Each replicate was tested on the same day, with one lot of reagents, by one operator, using one set of instruments. The variant calls in the spiked samples were compared to the control samples. The results are summarized in the following table:
| Haplotypes | No. of Replicates | No. of Correct Calls | No. of Incorrect Calls | No. of FQC | % of FQC | % Concordance with Control |
| --- | --- | --- | --- | --- | --- | --- |
| DQ2.2 | 12 | 12 | 0 | 0 | 0% | 100% |
| DQ2.5* | 24 | 24 | 0 | 0 | 0% | 100% |
| DQ7 | 12 | 12 | 0 | 0 | 0% | 100% |
| DQ8 | 12 | 12 | 0 | 0 | 0% | 100% |
| Total | 60 | 60 | 0 | 0 | 0% | 100% |
| *Includes two heterozygous samples | | | | | | |
No samples failed QC. All samples tested with the endogenous substances yielded 100% concordance with the control. None of the tested substances interfered with the GlutenID test at the concentrations tested.
ii. Exogenous Interference
Samples were self-collected by five individuals (representing the four celiac disease genetic risk haplotypes DQ2.2, DQ2.5, DQ7, and DQ8) at three collection times: before consuming an exogenous substance (baseline), immediately (0 minutes) after, and 30 minutes after. At each collection point, an aliquot of the saliva sample collected from
K241456 - Page 10 of 24
{10}
ORAcollect Dx sample collection device was processed according to the laboratory's SOP for DNA extraction. The testing conditions included:
- eating beef
- eating food other than beef
- drinking
- chewing gum
- using mouthwash
At each testing point, the samples were tested in triplicate, yielding a total of 225 results (5 donors x 5 conditions x 3 time points x 3 replicates). The combined results are summarized in the table below:
| Haplotypes | No. of Replicates | No. of Correct Calls | No. of Incorrect Calls | No. of FQC | % of FQC | % Concordance |
| --- | --- | --- | --- | --- | --- | --- |
| DQ2.2 | 45 | 44 | 0 | 1 | 2.2% | 100% |
| DQ2.5* | 90 | 85 | 0 | 5 | 5.6% | 100% |
| DQ7 | 45 | 45 | 0 | 0 | 0% | 100% |
| DQ8 | 45 | 44 | 0 | 1 | 2.2% | 100% |
| Total | 225 | 218 | 0 | 7 | 3.1% | 100% |
| *Includes two heterozygous samples | | | | | | |
Out of a total of 225 replicates run, 7/225 (3.1%) failed QC (FQC). All seven had LowDP (sequencing depth coverage below the laboratory cutoff for the assay) and were collected after exposure to exogenous interfering substances. Of the failures, five were seen with DQ2.5, one with DQ8 and one with DQ2.2. Two failures came immediately after eating beef (one with DQ2.5 and one with DQ8), one (DQ2.5) immediately after drinking and one (DQ2.5) immediately after mouthwash. Three of the failures occurred 30 minutes after exposure to beef (one DQ2.2), non-beef food (one DQ2.5) and chewing gum (one DQ2.5). All FQC samples were repeated and yielded correct results on the second run.
Based on these results, it is recommended that collection of saliva samples occurs at least 30 minutes after eating, drinking, using mouthwash, and brushing teeth.
iii. Smoking
A study was performed to evaluate the effects of smoking before saliva collection on the performance of the GlutenID Celiac GHR Test. Five individuals (representing the four haplotypes DQ2.2 DQ2.5, DQ7, and DQ8) provided three samples each (baseline/control sample taken at least 60 minutes prior to smoking, samples collected immediately after smoking, and samples collected 30 minutes after smoking. The smoking samples were tested in triplicate for a total of 45 test samples (5 donors x 3 time points x 3 replicates). The results are summarized in the following table:
K241456 - Page 11 of 24
{11}
| Haplotypes | No. of Replicates | No. of Correct Calls | No. of Incorrect Calls | No. of FQC | % of FQC | % of Concordance |
| --- | --- | --- | --- | --- | --- | --- |
| DQ2.2 | 9 | 9 | 0 | 0 | 0% | 100% |
| DQ2.5* | 18 | 15 | 0 | 3 | 16.7% | 100% |
| DQ7 | 9 | 9 | 0 | 0 | 0% | 100% |
| DQ8 | 9 | 9 | 0 | 0 | 0% | 100% |
| Total | 45 | 42 | 0 | 3 | 6.7% | 100% |
| *Includes two heterozygous samples | | | | | | |
Out of a total of 45 replicates run, three (6.7%) failed QC in this study. All three are with DQ2.5 and had DNA concentrations below LoQ (1 ng/μL) of the assay, and were collected immediately after smoking. These results indicate that samples saliva samples should be collected at least 30 minutes after smoking.
iv. Microbial Interference
A microbial interference study was performed to assess whether microbial DNA affects the performance of the GlutenID Celiac GHR Test. Five DNA samples representing the four tested haplotypes (DQ2, DQ8, DQ7, and DQ2.2) were spiked with and without DNA from the following five microbes:
- Lactobacillus casei
- Staphylococcus epidermidis
- Streptococcus mutans
- Aggregatibacter actinomycetemcomitans
- Candida albicans
All samples were tested in triplicate. The combined results are summarized in the following table:
| Haplotypes | No. of Replicates | No. of Correct Calls | No. of Incorrect Calls | No. of FQC | % of FQC | % of Concordance |
| --- | --- | --- | --- | --- | --- | --- |
| DQ2.2 | 15 | 15 | 0 | 0 | 0% | 100% |
| DQ2.5* | 30 | 30 | 0 | 0 | 0% | 100% |
| DQ7 | 15 | 15 | 0 | 0 | 0% | 100% |
| DQ8 | 15 | 15 | 0 | 0 | 0% | 100% |
| Total | 75 | 75 | 0 | 0 | 0% | 100% |
| *Includes two heterozygous samples | | | | | | |
No samples failed QC and all replicates produced correct genotype calls in the presence of microbial DNA, demonstrating no interference from the DNA from the tested microbes.
6. Assay Reportable Range:
Not applicable
K241456 - Page 12 of 24
{12}
7. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):
i. Reagent stability:
The GlutenID Celiac GHR Test uses the reagents for NGS to determine an individual’s GHR for celiac disease. The NGS sequencing reagents and related commercially available reagents should be stored under recommended conditions and used within the expiration date per the manufacturers’ instruction for use.
For PCR, a GlutenID Master Mix containing primers, polymerase and dNTPs are prepared and aliquoted by the lab and stored at -20°C. To determine the stability of the GlutenID Master Mix, three different lots of Master Mix (M1, M2, M3) were prepared from component reagents and stored at -20°C. The stability of the Master Mix was tested at 1, 4, 8, 12, 13 weeks and 6, 12, 18, 24 and 25 months. At each testing time point, three production lots were prepared by including each of three Master Mix lots for PCR and additional critical reagents for sequencing and library preparation (All critical reagents are stored at the recommended condition within their expiration date per the manufacturers’ instructions for use). At each designated timepoint, a 6-sample panel with haplotypes DQ2, DQ2+DQ2, DQ8, DQ8+DQ8, DQ2.2 (half-DQ2), DQ7 covering the six celiac genetic risk SNPs was tested in replicates of three for each sample across three lots, yielding a total of 54 results (6 samples x 3 replicates x 3 lots). The stability study is on-going; the already collected data supports the stability of the Master Mix up to 12 months at -20°C.
The stability of the aliquoted Master Mix under freeze/thaw cycles was evaluated. The results indicated that the aliquots of Master Mix are stable up to two freeze/thaw cycles.
ii. Stability of collection device and specimens:
Saliva samples are collected with the ORAcollect·Dx OCD-100. Refer to K152464 for pre-collection shelf-life stability of the collection device, stability of samples post-saliva collection, and freeze-thaw stability of samples stored in this collection device.
iii. Controls
The reagent performance used for the GlutenID Celiac GHR Test is verified daily by running, prior to use, the following quality control materials:
- Negative Control: Non-template control (NTC) with no DNA consisting of sterile water at the same volume as DNA samples and set up as an "unknown" sample in every run.
- Positive Control: Commercially purchased cell line which is positive for homozygous half-DQ2+half-DQ2 (DQ2.2+DQ2.2).
8. Detection Limit:
The Limit of Detection (LoD) of the GlutenID Celiac GHR Test was determined by testing six unique DNA samples representing haplotypes: DQ2, DQ2+DQ2, DQ8, DQ8+DQ8, DQ2.2+DQ7 and DQ2.2+DQ7. Each sample was tested at three DNA concentrations levels:
K241456 - Page 13 of 24
{13}
10 ng/μL, 5 ng/μL and 1 ng/μL, in replicates of two, using three different lots of reagents. The study yielded 36 results (6 samples x 2 replicates x 3 lots) per DNA concentration level and a total of 108 results (36 x 3 DNA levels). LoD was defined as the lowest DNA concentration at which at least 95% of samples resulted correct calls by the GlutenID Celiac GHR Test.
The results showed that, in a total of 108 replicates analyzed, no samples failed QC (FQC) and 100% correct genotype calls were observed for all samples, all reagent lots and all DNA concentration levels. The LoD of the GlutenID Celiac GHR Test is set as 1 ng/μL.
9. Assay Cut-Off:
Not applicable
B Comparison Studies:
1. Comparison with Sanger Bi-directional Sequencing:
The accuracy of the GlutenID Celiac GHR Test was assessed by evaluating a panel of samples that are heterozygous and homozygous for the reported haplotypes and covering all the possible results provided by the assay. A total of 143 samples representing all genetic variants tested by the assay were analyzed and compared with bi-directional Sanger sequencing (reference method) to calculate percent agreement with the sequencing results considered to be "truth." The number of samples for each haplotype is based on population frequencies reported in Ojnaga et al., (2018) Enferm Dig 110:421-426. These samples included 20 unique samples representing the wild-type genotype (homozygous common) and 123 samples positive for the four celiac risk haplotypes:
- HLA-DQ2.5 (rs2187668) wildtype (CC), heterozygous (CT), homozygous (TT)
- HLA-DQ8 (rs7454108) wildtype (TT), heterozygous (TC), homozygous (CC)
- HLA-DQ7 (rs4639334) wildtype (AA), heterozygous (AG), homozygous (GG)
- HLA-DQ2.2 (rs7775228) wildtype (GG), heterozygous (GT), homozygous (TT)
The percent agreement (PA) for each haplotype, the overall percent agreement and two-sided 95% CIs were calculated. The PAs per haplotype from clinical samples were calculated separately, and the results are summarized in the tables below:
| DQ2.5 |
| --- |
| Clinical Samples | Genotype | Sanger Bi-directional Sequencing |
| C/C | C/T | T/T |
| GlutenID Celiac GHR Test | C/C (homozygous common) | 20 | 0 | 0 |
| C/T (heterozygous) | 0 | 20 | 0 |
| T/T (homozygous rare) | 0 | 0 | 20 |
| FQC | 0 | 0 | 0 |
| Total=60 | 20 | 20 | 20 |
| PA (C/C | C/C) = 100% (20/20) with 95% CI: 83.9%-100%
PA (C/T | C/T) = 100% (20/20) with 95% CI: 83.9%-100%
PA (T/T | T/T) = 100% (20/20) with 95% CI: 83.9%-100%
Percent of FQC is 0.0% (0/60) 95% CI: 0.0%-6.0% |
K241456 - Page 14 of 24
{14}
| DQ8 |
| --- |
| Clinical Samples | Genotype | Sanger Bi-directional Sequencing |
| T/T | T/C | C/C |
| GlutenID Celiac GHR Test | T/T (homozygous common) | 20 | 0 | 0 |
| T/C (heterozygous) | 0 | 20 | 0 |
| C/C (homozygous rare) | 0 | 0 | 3 |
| FQC | 0 | 0 | 0 |
| Total=43 | 20 | 20 | 3 |
| PA (T/T | T/T) = 100% (20/20) with 95% CI: 83.9%-100%
PA (T/C | T/C) = 100% (20/20) with 95% CI: 83.9%-100%
PA (C/C | C/C) = 100% (3/3) with 95% CI: 43.9%-100%
Percent of FQC is 0.0% (0/43) with 95% CI: 0.0%-8.2% |
| DQ7 |
| --- |
| Clinical Samples | Genotype | Sanger Bi-directional Sequencing |
| A/A | A/G | G/G |
| GlutenID Celiac GHR Test | A/A (homozygous common) | 20 | 0 | 0 |
| A/G (heterozygous) | 0 | 20 | 0 |
| G/G (homozygous rare) | 0 | 0 | 10 |
| FQC | 0 | 0 | 0 |
| Total=50 | 20 | 20 | 10 |
| PA (A/A | A/A) = 100% (20/20) with 95% CI: 83.9%-100%
PA (A/G | A/G) = 100% (20/20) with 95% CI: 83.9%-100%
PA (G/G | G/G) = 100% (10/10) with 95% CI: 72.3%-100%
Percent of FQC is 0.0% (0/50) with 95% CI: 0.0%-7.1% |
| DQ2.2 (rs7775228) |
| --- |
| Clinical Samples | Genotype | Sanger Bi-directional Sequencing |
| G/G | G/T | T/T |
| GlutenID Celiac GHR Test | G/G (homozygous common) | 20 | 0 | 0 |
| G/T (heterozygous) | 0 | 20 | 0 |
| T/T (homozygous rare) | 0 | 0 | 10 |
| FQC | 0 | 0 | 0 |
| Total=50 | 20 | 20 | 10 |
| PA (G/G | G/G) = 100% (20/20) with 95% CI: 83.9%-100%
PA (G/T | G/T) = 100% (20/20) with 95% CI: 83.9%-100%
PA (T/T | T/T) = 100% (10/10) with 95% CI: 72.3%-100%
Percent of FQC is 0.0% (0/50) with 95% CI: 0.0%-7.1% |
Calculation of PA per allelic variant between the GlutenID Celiac GHR Test results and bidirectional Sanger sequencing was 100% concordant for heterozygous and homozygous
K241456 - Page 15 of 24
{15}
DQ2.5, DQ8, DQ7, DQ2.2 haplotypes. The results showed 100% (203/203) of concordance between GlutenID Celiac GHR test results and the Sanger sequencing method (the 95%CI is 98.1%–100%). The percentage of FQC per allelic variant was also calculated and was 0% (0/203) with 95%CI: (0.0%–1.9%). The study showed that the GlutenID Celiac GHR Test was able to correctly genotype all samples analyzed.
Percentages of variants in the target population as reported by Liu et al. is presented in the table below:
| Variant | White N (%) | African American N (%) | Asian N (%) | Hispanic, Biracial, & Others N (%) | Total |
| --- | --- | --- | --- | --- | --- |
| DQ2+DQ2 | 250 (1.4) | 28 (1.2) | 1 (0.2) | 80 (0.6) | 359 (1.1) |
| DQ2+DQ8 | 415 (2.4) | 30 (1.8) | 4 (0.7) | 254 (2.2) | 703 (2.2) |
| ^DQ2+half-DQ2 | None | | | | |
| DQ8+DQ8 | 385 (2.2) | 6 (0.3) | 5 (0.9) | 28 (2.7) | 703 (2.2) |
| ^DQ8+half-DQ2 | None | | | | |
| ^half-DQ2+half-DQ2 | | | | | |
| ^DQ7+half-DQ2 DQ2(trans) | | | | | |
| *DQ2.5+DQX DQ2(cis) | 3376 (19.2) | 472 (20.6) | 68 (11.8) | 1530 (13.5) | 5446 (17.1) |
| *DQ8+DQX | 2803 (15.9) | 172 (7.5) | 51 (8.9) | 2613 (23.1) | 5639 (17.8) |
| ^half-DQ2 | None | | | | |
| ^DQ7 | | | | | |
| Non-celiac genetics (NCG) | 10,386 (59.0) | 1582 (69.1) | 447 (77.6) | 6501 (57.6) | 18,916 (59.5) |
| Total | 17,615 | 2290 | 576 | 11,285 | 31,766 |
| Diplotype combinations are represented by the following symbolic nomenclature:
*DQX is any haplotype except DQ2.5, DQ8
^The DQ2(trans) haplotypes (DQ2.2 and DQ7) representing 1.4-5.3% of the general population were not included in the study.
Prevalence of DQ2(trans) is <5% of all patients with celiac disease. | | | | | |
Technical (analytical) positive predictive values for GlutenID Celiac GHR Test results were >92.4%.
2. Matrix Comparison:
Not applicable
C Clinical Studies:
1. Clinical Sensitivity and Clinical Specificity:
Not applicable
2. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):
K241456 - Page 16 of 24
{16}
a. Disease Association and Pathogenesis
The pathophysiology of celiac disease involves a systemic, abnormal immune response to dietary gluten in genetically predisposed individuals. Immune cells attack the small intestine and other tissues causing symptoms such as diarrhea, malabsorption, and anemia.
HLA-DQA1 and HLA-DQB1 genes encode the alpha and beta chains of the HLA-DQ2 and HLA-DQ8 heterodimeric Major Histocompatibility complex (MHC) Class II surface proteins expressed on the surface of antigen presenting cells. MHC Class II proteins on the surface of dendritic cells and monocytes present antigens to immune system T-cells initiating response to foreign antigens. Gluten-reactive T-cells can cause chronic inflammation of the small intestine with accompanying villus atrophy leading to malabsorption. The HLA-DQ2.5 haplotype is present in 95% of patients diagnosed with celiac disease and the HLA-DQ8 haplotype is detected in <5% (Taylor 2015¹). Rarely, HLA-DQ2 is inherited in the form of two separate haplotypes HLA-DQ7 and HLA-DQ2.2 (one half from each parent) encoding alpha and beta chains differing by only two amino acids from DQ2.5 and conferring similar celiac disease risk (Pisapia 2020²). The DQ2.5 haplotype is commonly referred to as DQ2(cis) while the DQ7+DQ2.2 haplotype is called DQ2(trans). Although a hetero or homozygous DQ7 haplotype inherited without DQ2.2 confers only slightly increased risk above non celiac genetics (NCG), laboratory testing for celiac genetic risk must include all four haplotypes (DQ2.5, DQ8, DQ2.2, and DQ7) to identify presence or absence of DQ2(trans).
b. Risk Categorization
Depending on the specific variant combination detected, the GlutenID Celiac Genetic Health Risk Test provides the individuals' genetic health risk for developing celiac disease. Personalized reports are generated for each consumer that help users understand the meaning of their results and any appropriate next steps. The GlutenID Celiac GHR Test uses three categories to define risk. One of these risk categories has been assigned to each genetic result. The risk categorization is based on the reported clinical cases (published references) for each genetic result.
The following three categories are reported for the GlutenID test:
- Increased Risk: Estimated post-test risk is higher than general population risk of 1%.
- Low Risk: Estimated post-test risk is the same or lower than the general population risk of 1%.
- Not likely at Risk: Estimated post-test risk is 0.5% or below.
The GlutenID Celiac Genetic Health Risk (GHR) Test is indicated for detecting the presence or absence of four HLA-DQ haplotype variants associated with the risk of
K241456 - Page 17 of 24
¹ Taylor et al., (2019). Celiac Disease. GeneReviews [Internet] https://www.ncbi.nlm.nih.gov/books/NBK1727
² Pisapia et al., (2020). Differential expression of pre-disposing HLA-DQ2.5 alleles in DR5/DR7 celiac disease patients affects the pathological immune response to gluten. Nature Sci Rep 10:17227 https://doi.org/10.1038/s41598-020-73907-2
{17}
developing celiac disease. The GlutenID Celiac GHR Test reports variants associated with the HLA-DQ2.5 haplotype, the HLA-DQ8 haplotype, the HLA-DQ7 haplotype, and the HLA-DQ2.2 haplotype. Inheritance of one or more of these haplotypes is associated with higher risk for development of Celiac disease (Megiorni 2009³). The GlutenID test uses SNP genotyping to identify four HLA haplotype variants that confer risk for celiac disease (Monsuur 2008⁴).
Likelihood ratios (LR) for each test result were evaluated based on published literature. The confidence intervals for the LR were calculated using the asymptotic method for the ratios of two independent binomial proportions (see CLSI EP12). For each test result, post-test risk (R) was calculated based on the corresponding LR and pre-test risk π as R/(1-R) = LR × π/(1-π), where π is pre-test risk of 1%.
In the clinical study described in F. Megiorni, B. Mora, M. Bonamico, M. Barbato, R. Nenna, G. Maiella, P. Lulli, M. Mazzilli “HLA-DQ and risk gradient for celiac disease” in Human Immunology 70 (2009), there were 437 subjects with celiac disease, and 551 subjects without celiac disease. Estimations of the LR for different GlutenID Celiac Genetic Health Risk (GHR) Test results were calculated and presented in the table below:
| Test Result | LR | 95% CI for LR | Post-Test Risk for Pre-Test Risk of 1% | Risk Category Reported by Test |
| --- | --- | --- | --- | --- |
| DQ2+DQ2 | 9.8 | (5.6; 17.1) | 8% | Increased |
| DQ2+half-DQ2 | | | | |
| DQ2+DQ8 | 13.9 | (2.3; 83.5) | 6% | Increased |
| DQ8+half-DQ2 | 4.1 | (1.4; 11.9) | 4% | Increased |
| DQ2+DQ7 | 2.9 | (2.4; 3.5) | 3% | Increased |
| DQ2.2+DQ7 (DQ2 trans) | | | | |
| DQ2 (cis) | | | | |
| DQ8+DQ8 | 1.1 | (0.7; 1.8) | 1% | Low |
| DQ8 | | | | |
| DQ8+DQ7 | | | | |
| Half-DQ2+Half-DQ2 (DQ2.2+DQ2.2) | 0.48 | (0.29; 0.78) | 0.5% | Not Likely at Risk |
| Half-DQ2(DQ2.2) | | | | |
| DQ7+DQ7 | | | | |
| DQ7 | | | | |
| Non-celiac genetics (NCF) | 0.05 | (0.03; 0.08) | 0.05% | Not Likely at Risk |
Individuals who are homozygous for DQ2(cis) are at highest risk for development of celiac disease (Bajor 2019⁵). Conversely individuals who inherit either DQ7 or DQ2.2
K241456 - Page 18 of 24
{18}
(but not both) are at lower risk for celiac disease but have 50% chance of transmitting a risk allele to future generations.
Historically, the medical literature has focused primarily on celiac disease genetics in individuals of European ancestry where an estimated 25% of this population carries at least one copy of the HLA-DQ2.5 haplotype (Singh 2018⁶). Although celiac disease is a polygenic disorder, over 90% of patients with celiac disease have at least one copy of the HLA-DQ2.5 haplotype.
3. User Studies
a. Saliva collection kit usability study:
Refer to K212745 for the saliva collection device (ORAcollect·Dx OCD-100) instructions for use and to assess the ability of lay users to provide samples adequate for testing.
b. GlutenID Celiac GHR Test user comprehension study:
**Objectives:**
The user comprehension study was performed to assess user comprehension of the GlutenID Celiac GHR Test reports. The study was performed with a demographically diverse group of study participants 18 years old or older by asking questions for five comprehension categories (purpose of test, limitations, relevant ethnicities, meaning of results, other factors that may have an impact on test results, and appropriate action) of the GlutenID Celiac GHR Test reports.
**Methods:**
Quota-based sampling per U.S. Census was employed to recruit a diverse set of study participants based on their demographic variables in age, gender, race/ethnicity, education level and geographic region. Study participants who enrolled into the user comprehension study and passed the screening questionnaire to ensure that they met the inclusion criteria for the study were randomly assigned into one of three risk category groups ('Not likely at risk', 'Low risk', and 'Increased risk'). Exclusion criteria were applied in the recruitment stage, as well as during the study data collection. All exclusions were counted and documented during the study and were reported with a clear description of each excluded participants' demographics and detailed reason for exclusion. During the analysis, the demographics of all participants (defined as those who consent to participate) was compared to the demographics of those remaining after exclusion, to evaluate whether the exclusion process potentially introduced any bias.
Of the 15 total possible GHR Reports that can be generated by the GlutenID Celiac GHR Test, three were selected for the User Comprehension Study. The selected Reports summarized in the table below cover all the possibilities for number of variants and risk
K241456 - Page 19 of 24
⁶ Singh et al., (2018). Global prevalence of celiac disease: systematic review and meta-analysis. Clin Gastroenterol and Hepatol. 16:823-83.
{19}
categories associated with celiac disease that can be reported by the GlutenID Celiac GHR Test:
- Number of haplotypes detected: a total of three different reports can be obtained: 0, 1, or 2 haplotypes detected.
- Risk category: three different risk categories ('Not likely at risk', 'Low risk', and 'Increased risk') can be provided by the test, which represent the risk for developing celiac disease.
| | GHR Report | Number of Haplotypes Detected | Risk Category for Celiac Disease |
| --- | --- | --- | --- |
| 1 | GlutenID NCG* | 0 | Not likely at risk |
| 2 | GlutenID DQ2cis | 1 | Low risk |
| 3 | GlutenID DQ2+DQ8 | 2 | Increased risk |
*Non-Celiac Genetic
As the GHR Report for 'Variant not determined' was not included in the User Comprehension Study, the 'No Results' email text that the user will receive when the laboratory is unable to complete DNA testing due to insufficient or no human DNA being present in the collected saliva sample was also shown to all study participants along with two questions designed to evaluate whether the key concepts are communicated effectively. Each GHR Report includes the following sections: 'Your Result' (Result Report), 'Scientific Details', 'Frequently Asked Questions', and 'Glossary of Terms'
User comprehension was tested through a two-step process. First, participants' comprehension was tested prior to viewing the Educational Module, and GHR Reports. The Pre-Test questionnaire included basic demographics (for inclusion/exclusion criteria) and baseline comprehension questions that covered the following comprehension domains: test purpose [PURPOSE], test limitations [LIMITATIONS], race/ethnicity [ETHNICITY], and other factors that may have an impact on the test results [OTHER FACTORS]. Second, on completion of reviewing a representative sample of all the material that is included in the Education Module and the GlutenID reports, participants were provided with the Post-Test questionnaire to complete (approximately 20 minutes). The Post-Test questionnaire included comprehension questions designed to measure all six comprehension domains: test results [RESULTS] and appropriate action [NEXT STEPS] in addition to PURPOSE, LIMITATIONS, ETHNICITY, OTHER FACTORS. The RESULTS and NEXT STEPS domains were only included in the Post-Test questionnaire because they represented specific results that participants could not know prior to viewing the GHR reports and related materials. Each comprehension domain included at least two comprehension questions.
The GHR Reports and Supplemental Materials were reviewed by a Certified Genetic Counselor (GC) to ensure that the materials sufficiently cover the relevant general and variant-specific concepts [comprehension domains]. Prior to recruitment for the user comprehension study, the "No Result" email and test report materials (GHR Report, Education Module, and Supplemental Materials), with content targeted no more difficult than $8^{\text{th}}$ grade reading level, were evaluated for readability.
K241456 - Page 20 of 24
{20}
A total of 361 participants were engaged in this study. Of the 361 participants, 18 were determined to be ineligible and qualified as exclusions, leaving 343 eligible participants in the study. Of those eligible, a total of 3 failed to complete the Pre-Test. A total of 340 completed the Pre-Test and were invited to participate in the POST survey. A total of 26 failed to complete the Post-Test, leaving 314 complete eligible participants at study completion. This means the study had a completion rate of 92%. There were no statistical differences observed between those who consented (n=361) and those who eventually participated and completed the study (post-exclusion; n=314).
The study was set up to randomly assign each eligible study participant to one of three study arms identified by the three pre-selected test report types, with the following number of eligible participants that successfully completed the User Comprehension Study as summarized in the table below for each study arm.
| Study Arm | GHR Report | Total Participants Completed Study |
| --- | --- | --- |
| 1 | GlutenID NCG* | 109 |
| 2 | GlutenID DQ2(cis) | 101 |
| 3 | GlutenID DQ2+DQ8 | 105 |
| Total | | 314 |
*Non-Celiac Genetic
## Results
Comprehension scores were calculated using the total number of responses received that were correct over the total number of eligible responses received. The calculated proportion for each domain serves as the comprehension score for that domain. The target score for each comprehension domain was 90% across all reports. A mean comprehension rate was calculated across all comprehension question domains and all reports. To evaluate whether the comprehension scores had significantly improved statistically, the pre- and post-test scores in each domain, and overall, were compared by using paired sample t-test. Results summarized in the table below show that each comprehension domain achieved a minimum of 79.3% or higher user comprehension score in the pre-test questionnaire, and 94.0% or higher user comprehension score in the post-test questionnaire, across all reports.
| User Comprehension Pre- to Post-Test Scores for all GHR Reports by Domain | | | | |
| --- | --- | --- | --- | --- |
| Domain Name | N | Pre-Test (%) | Post-Test (%) | Improvement (%) |
| Education Module | 314 | 88.2 | 97.8 | 9.6 |
| Purpose | 314 | 95.1 | 97.7 | 2.6 |
| Other Factors | 314 | 94.8 | 96.7 | 1.9 |
| Limitations | 314 | 80.2 | 95.9 | 15.7 |
| Race/Ethnicity | 314 | 79.3 | 96.8 | 17.5 |
| Average of all domains | 314 | 87.5 | 97.0 | 9.5 |
K241456 - Page 21 of 24
{21}
| User Comprehension Pre- to Post-Test Scores for all GHR Reports by Domain | | | | |
| --- | --- | --- | --- | --- |
| Domain Name | N | Pre-Test (%) | Post-Test (%) | Improvement (%) |
| Results | 314 | N/A | 94.4 | N/A |
| Next Steps | 314 | N/A | 96.1 | N/A |
| | | | | |
| Variant not determined^ | 314 | N/A | 99.5 | N/A |
^ The lab is unable to complete the DNA tests due to insufficient or no human DNA being present in the collected sample
The overall comprehension scores were of 87.5% and 97.0% across all comprehension domains (i.e., Education Module, Purpose, Other Factors, Limitations and Race/Ethnicity) for the pre-test and post-test questionnaire, respectively. Results of the pre- to post-test comparison by paired sample t-tests showed a statistically significant improvement in the user comprehension scores between the pre-and post-test questionnaire for the Education Module, and the domains of 'Limitations', and 'Ethnicity'. Statistical significance demonstrates that the increase in the user comprehension score was a result of the education material. The 'Purpose' and 'Other Factors' domains showed no significant difference.
When analyzed individually for each of the three GHR reports, the post-test comprehension scores for each domain are 91% or higher (Next Steps domain in the NCG (GlutenID Non-Celiac Genetic report has a post-test score of 91%) as shown in the table below.
| Post-Test Comprehension Rates (%) by GHR Report Type | | | |
| --- | --- | --- | --- |
| Domain Name | GlutenID
NCG* | GlutenID
DQ2(cis) | GlutenID
DQ2+DQ8 |
| Education | 97 | 99 | 97 |
| Purpose | 97 | 98 | 98 |
| Other Factors | 97 | 97 | 97 |
| Limitations | 94 | 95 | 99 |
| Ethnicity | 97 | 97 | 97 |
| Results | 97 | 94 | 97 |
| Next Steps | 91 | 94 | 98 |
| Average of all Domains | 96 | 96 | 98 |
| | | | |
| Variant not determined^ | 99 | 100 | 100 |
* Non-Celiac Genetic
^ The lab is unable to complete the DNA tests due to insufficient or no human DNA being present in the collected sample
The following table summarizes the user comprehension scores (post-test only) broken out by demographic category (age, sex, race/ethnicity, education level, and geographic region) per comprehension domain for the overall study (all three GHR reports combined). Every demographic category scored 92% or higher in every comprehension domain across all report types. Results showed that participant demographics were unrelated to user comprehension.
K241456 - Page 22 of 24
{22}
| User Comprehension Post-Test Scores for all Reports by Demographic | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | Comprehension Domain | | | | | | | |
| | | Education | Purpose | Other Factors | Limitations | Ethnicity | Next Steps | Results | Variant not determined |
| | N | Mean (%) | | | | | | | |
| Overall | 314 | 98 | 98 | 97 | 96 | 97 | 96 | 94 | 100 |
| Age | | | | | | | | | |
| 18-34 | 101 | 98 | 98 | 99 | 95 | 97 | 96 | 95 | 100 |
| 35-49 | 112 | 98 | 99 | 97 | 96 | 98 | 97 | 96 | 100 |
| 50+ | 101 | 97 | 96 | 94 | 96 | 96 | 96 | 92 | 99 |
| Sex^ | | | | | | | | | |
| Male | 161 | 97 | 98 | 96 | 96 | 96 | 95 | 94 | 100 |
| Female | 152 | 99 | 98 | 97 | 96 | 97 | 97 | 95 | 99 |
| Race/Ethnicity* | | | | | | | | | |
| White Alone | 184 | 99 | 98 | 98 | 96 | 97 | 97 | 94 | 99 |
| Non-White | 130 | 97 | 97 | 96 | 95 | 96 | 95 | 95 | 100 |
| Education** | | | | | | | | | |
| Some College | 152 | 98 | 97 | 96 | 96 | 97 | 97 | 95 | 99 |
| Bachelor's Degree or Higer | 162 | 97 | 98 | 97 | 96 | 97 | 95 | 94 | 100 |
| Geographic Region | | | | | | | | | |
| Northeast | 69 | 97 | 97 | 97 | 98 | 97 | 96 | 94 | 99 |
| Midwest | 67 | 97 | 96 | 97 | 98 | 97 | 96 | 92 | 99 |
| South | 74 | 98 | 97 | 96 | 94 | 97 | 95 | 96 | 100 |
| West | 104 | 99 | 99 | 97 | 98 | 97 | 95 | 96 | 100 |
^ one participant identified as non-binary or other "not listed" gender
* Non-White: include African American, Asian American, Middle Eastern/North African, Native American, Native Hawaiian/Other Pacific Islander, biracial or Multiracial, and Hispanic
** Some college: include associate degree or less
c. Frequently Asked Questions (FAQ) Material:
A Frequently Asked Questions (FAQs) section was developed and included in the GlutenID Celiac GHR Test report. The FAQs section provides users with information to adequately understand the purpose, limitations, and meaning of results of the test and was developed to be consistent with the manufacturer's labeling design, identification of communication messages, and label comprehension. The concepts covered in the FAQs include: test results, meaning of test results, purpose of the test, test limitations, other risk factors that contribute to disease, relevance of race and ethnicity on test results, how the test results may affect the user's family and children, appropriate follow-up procedures, links to resources for additional information.
The questions included in the FAQ section for each of the test report included the following:
- What is celiac disease?
- Can celiac disease affect anyone?
K241456 - Page 23 of 24
{23}
- How is celiac disease treated?
- What is the purpose of the GlutenID test?
- How accurate is the GlutenID test?
- Does the GlutenID test look at my whole genetic profile?
- My report says I have NONE of the celiac risk variants, what does this mean?
- My result says, ‘Not Likely at Risk’, what next steps should I take?
- Based on my results, should I immediately remove gluten from my diet?
- What are the limitations of the GlutenID test?
- How does this result affect my family?
- How does a person’s ethnic background impact the results?
- If I am not of European descent does the test apply to me?
- Where can I find more information?
- How do I get an appointment with a genetic counselor?
Each GlutenID Celiac Genetic Health Report provides information to the FAQs that are specific to celiac disease risk haplotypes.
d. User Opt-In page:
Not applicable
**D Clinical Cut-Off:**
Not applicable
**E Expected Values/Reference Range:**
Not applicable
**VIII Proposed Labeling:**
The labeling supports the finding of substantial equivalence for this device.
**IX Conclusion:**
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
K241456 - Page 24 of 24
Panel 1
/
Sort by
Ready
Predicate graph will load when search results are available.
Embedding visualization will load when search results are available.
PDF viewer will load when search results are available.
Loading panels...
Select an item from Submissions
Click any panel, subpart, regulation, product code, or device to see details here.
Section Matches
Results will appear here.
Product Code Matches
Results will appear here.
Special Control Matches
Results will appear here.
Loading collections...
Loading
My Alerts
You will receive email notifications based on the filters and frequency you set for each alert.
Sort by:
Create Alert
Search Filters
Agent Token
Create a read-only bearer token for Claude, ChatGPT, or other agents that can call HTTP APIs.
