title: High-throughput beta-thalassemia carrier screening by allele-specific Q-primer real-time polymerase chain reaction.
abstract: Based on a novel Q-primer real-time polymerase chain reaction (PCR) system, we designed allele-specific Q-primers for the detection of three beta-thalassemia mutations [Cd41/42(-TCTT), IVSI nt5 (G>C), and IVSII nt654 (C>T)] that have a high carrier frequency in Southeast Asia. With clear distinction between heterozygote and wild-type, DeltaC(t) (threshold cycle) values were defined. The results of evaluating 139 blinded samples by our system match perfectly with those obtained by the conventional reverse dot blot (RDB) method. With a 384-well plate that included replicates in the same analysis, our throughput reached 190 reactions per run with a turnaround time as short as 130 min, and the cost of consumables was as low as $1 (US) for each test.
authors.affiliation: National University of Singapore, Singapore.
authors.forename: Xiaokun
authors.initials: X
authors.lastname: Liu
authors.forename: Hai Yang
authors.initials: HY
authors.lastname: Law
authors.forename: Yuen Ming
authors.initials: YM
authors.lastname: Tan
authors.forename: Yan
authors.initials: Y
authors.lastname: Hong
mesh_terms: D000483:Alleles; D017931:DNA Primers / Q000737:chemistry*; D006580:Genetic Carrier Screening / Q000379:methods*; D006579:Heterozygote; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D017086:beta-Thalassemia / Q000235:genetics*
chemical_list: D017931:DNA Primers