Development and Characterization of Reference Materials for MTHFR, SERPINA1, RET, BRCA1, and BRCA2 Genetic Testing

Abstract

Well-characterized reference materials (RMs) are integral in maintaining clinical laboratory quality assurance for genetic testing. These RMs can be used for quality control , monitoring of test performance, test validation , and proficiency testing of DNA-based genetic tests. To address the need for such materials, the Centers for Disease Control and Prevention established the Genetic Testing Reference Material Coordination Program (GeT-RM), which works with the genetics community to improve public availability of characterized RMs for genetic testing. To date , the GeT-RM program has coordinated the characterization of publicly available genomic DNA RMs for a number of disorders, including cystic fibrosis, Huntington disease, fragile X, and several genetic conditions with relatively high prevalence in the Ashkenazi Jewish population. Genotypic information about a number of other cell lines has been collected and is also available. The present study includes the development and commutability/genotype characterization of 10 DNA samples for clinically relevant mutations or sequence variants in the following genes: MTHFR; SER-PINA1; RET; BRCA1; and BRCA2. DNA samples were analyzed by 19 clinical genetic laboratories using a variety of assays and technology platforms. Concordance was 100% for all samples, with no differences observed between laboratories using different methods. All DNA samples are available from Coriell Cell Repositories and characterization information can be found on the GeT-RM website.

Molecular diagnostics: harmonization through reference materials, documentary standards and proficiency testing

“…Gaucher disease, mucolipidosis IV, Neimann-Pick disease and Tay-Sachs disease) [22], cystic fibrosis [23], Huntington's disease [24], methylene tetrahydrofolate reductase-related homocysteinemia, a1-anti trypsin deficiency, multiple endocrine neoplasia and BRCA1-and BRCA2-related cancers [25]. Genomic DNA material was tested in between three and ten clinical genetic laboratories for each of these disorders using a variety of genetic assays, including DNA sequence analysis.…”

Utility of NIST Whole-Genome Reference Materials for the Technical Validation of a Multigene Next-Generation Sequencing Test

“…2 The first major limitation is the lack of controls for genes in rare conditions, whereas another issue is the availability of having a source of well-characterized, consistent, and renewable controls for each DNA variant tested. 5,6 A similar problem exists with proficiency testing for genetic laboratories, where the limitations of analyte-specific testing in the NGS era has led the College of American Pathologists to support the move toward a methods-based proficiency testing strategy. The College of American Pathologists concluded that a methods-based approach allows the standardization of proficiency testing among laboratories that use vastly different NGS panels and provides a practical, efficient, cost-effective, and thorough mechanism to administer testing.…”

Nucleic acid reference materials (NARMs): definitions and issues

“…The Center for Disease Control (CDC) has developed a process whereby the commutability of readily available materials have been tested in numerous laboratories (13). Additionally, the rapid evolution of technologies and methodologies used in molecular diagnostic testing exceeds the capacity to generate reference materials.…”