title: Development and performance of a comprehensive targeted sequencing assay for pan-ethnic screening of carrier status.
abstract: Identifying individuals as carriers of severe disease traits enables informed decision making about reproductive options. Although carrier screening has traditionally been based on ethnicity, the increasing ethnic admixture in the general population argues for the need for pan-ethnic carrier screening assays. Highly multiplexed mutation panels allow for rapid and efficient testing of hundreds of mutations concurrently. We report the development of the Pan-Ethnic Carrier Screening assay, a targeted sequencing assay for routine screening that simultaneously detects 461 common mutations in 91 different genes underlying severe, early-onset monogenic disorders. Mutation selection was aided by the use of an extensive mutation database from a clinical laboratory with expertise in newborn screening and lysosomal storage disease testing. The assay is based on the Affymetrix GeneChip microarray platform but generates genomic DNA sequence as the output. Analytical sensitivity and specificity, using genomic DNA from archived control cultures and from clinical specimens, was found to be >99% for all mutation types. This targeted sequencing assay has advantages over multiplex PCR and next-generation sequencing assays, including accuracy of mutation detection over a range of mutation types and ease of analysis and reporting of results.
authors: Emory Genetics Laboratory, Department of Human Genetics, Emory University, Atlanta, Georgia.
authors: Alice K
authors: AK
authors: Tanner
authors: Emory Genetics Laboratory, Department of Human Genetics, Emory University, Atlanta, Georgia.
authors: C Alexander
authors: CA
authors: Valencia
authors: Emory Genetics Laboratory, Department of Human Genetics, Emory University, Atlanta, Georgia.
authors: Devin
authors: D
authors: Rhodenizer
authors: Emory Genetics Laboratory, Department of Human Genetics, Emory University, Atlanta, Georgia.
authors: Marina
authors: M
authors: Espirages
authors: Emory Genetics Laboratory, Department of Human Genetics, Emory University, Atlanta, Georgia.
authors: Cristina
authors: C
authors: Da Silva
authors: TessArae LLC, Potomac Falls, Virginia.
authors: Lisa
authors: L
authors: Borsuk
authors: TessArae LLC, Potomac Falls, Virginia.
authors: Sara
authors: S
authors: Caldwell
authors: TessArae LLC, Potomac Falls, Virginia.
authors: Edward
authors: E
authors: Gregg
authors: TessArae LLC, Potomac Falls, Virginia.
authors: Elizabeth
authors: E
authors: Grimes
authors: TessArae LLC, Potomac Falls, Virginia.
authors: Agnieszka M
authors: AM
authors: Lichanska
authors: TessArae LLC, Potomac Falls, Virginia.
authors: Leah
authors: L
authors: Morris
authors: TessArae LLC, Potomac Falls, Virginia.
authors: Anjan
authors: A
authors: Purkayastha
authors: TessArae LLC, Potomac Falls, Virginia.
authors: Brian
authors: B
authors: Weslowski
authors: TessArae LLC, Potomac Falls, Virginia.
authors: Clark
authors: C
authors: Tibbetts
authors: TessArae LLC, Potomac Falls, Virginia.
authors: Matthew C
authors: MC
authors: Lorence
authors: Emory Genetics Laboratory, Department of Human Genetics, Emory University, Atlanta, Georgia. Electronic address: mhegde@emory.edu.
authors: Madhuri
authors: M
authors: Hegde
mesh_terms: D000328:Adult; D004252:DNA Mutational Analysis / Q000379:methods; D005006:Ethnicity / Q000235:genetics*; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D006579:Heterozygote; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*