title: High-throughput carrier screening using TaqMan allelic discrimination.
abstract: Members of the Ashkenazi Jewish community are at an increased risk for inheritance of numerous genetic diseases such that carrier screening is medically recommended. This paper describes the development and evaluation of 30 TaqMan allelic discrimination qPCR assays for 29 mutations on 2 different high-throughput platforms. Four of these mutations are in the GBA gene and are successfully examined using short amplicons due to the qualitative nature of TaqMan allelic discrimination. Two systems were tested for their reliability (call rate) and consistency with previous diagnoses (diagnostic accuracy) indicating a call rate of 99.04% and a diagnostic accuracy of 100% (+/-0.00%) from one platform, and a call rate of 94.66% and a diagnostic accuracy of 93.35% (+/-0.29%) from a second for 9,216 genotypes. Results for mutations tested at the expected carrier frequency indicated a call rate of 97.87% and a diagnostic accuracy of 99.96% (+/-0.05%). This study demonstrated the ability of a high throughput qPCR methodology to accurately and reliably genotype 29 mutations in parallel. The universally applicable nature of this technology provides an opportunity to increase the number of mutations that can be screened simultaneously, and reduce the cost and turnaround time for accommodating newly identified and clinically relevant mutations.
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway, New Jersey, United States of America. afedick@rmanj.com
authors.forename: Anastasia
authors.initials: A
authors.lastname: Fedick
authors.forename: Jing
authors.initials: J
authors.lastname: Su
authors.forename: Chaim
authors.initials: C
authors.lastname: Jalas
authors.forename: Lesley
authors.initials: L
authors.lastname: Northrop
authors.forename: Batsal
authors.initials: B
authors.lastname: Devkota
authors.forename: Josef
authors.initials: J
authors.lastname: Ekstein
authors.forename: Nathan R
authors.initials: NR
authors.lastname: Treff
mesh_terms: D000483:Alleles; D004252:DNA Mutational Analysis / Q000379:methods*; D005260:Female; D006580:Genetic Carrier Screening; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D007585:Jews; D008297:Male; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results
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