title: Novel technique for rapid detection of alpha-globin gene mutations and deletions.
abstract: Populations in Southeast Asia and South China have high frequencies of alpha-thalassemia caused by alpha-globin gene mutations and/or deletions. This study was designed to find an efficient and simple diagnostic test for the mutations and deletions. A duplex polymerase chain reaction (PCR)/denaturing high-pressure liquid chromatography (DHPLC) was used to detect the mutations and deletions. A blinded study of 110 samples, which included 92 alpha-thalassemia samples with various genotypes and 18 normal DNA samples, was carried out by the methods. The duplex PCR products of the sample with known Constand spring mutation (CS)/alphaalpha, Quonsze mutation (QS)/alphaalpha, and Weastmead mutation (WS)/alphaalpha DNA showed significantly different profiles, which suggests that DHPLC analysis at 63.8 degrees C can detect potential mutations directly. The DHPLC at 50 degrees C analysis can distinguish the --SEA and nondeletional alleles. The new assay is 100% concordant with the original genotype. In conclusion, the technique including the duplex PCR assay followed by DHPLC analysis can be used to diagnose alpha-thalassemia; this methodology is simple, rapid, accurate, semiautomatic, and high output, and thus, it is suitable for large-scale screening.
authors.affiliation: Basic Medical Research Center, Beijing Chaoyang Hospital, Affiliate of The Capital Medical University, Beijing, China 100020. liujingzhong090525@hotmail.com
authors.forename: Jingzhong
authors.initials: J
authors.lastname: Liu
authors.forename: Xingyuan
authors.initials: X
authors.lastname: Jia
authors.forename: Ning
authors.initials: N
authors.lastname: Tang
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
authors.forename: Xiaoyi
authors.initials: X
authors.lastname: Wu
authors.forename: Ren
authors.initials: R
authors.lastname: Cai
authors.forename: Lirong
authors.initials: L
authors.lastname: Wang
authors.forename: Quanzhang
authors.initials: Q
authors.lastname: Liu
authors.forename: Bai
authors.initials: B
authors.lastname: Xiao
authors.forename: Jim
authors.initials: J
authors.lastname: Zhu
authors.forename: Qingtao
authors.initials: Q
authors.lastname: Wang
mesh_terms: D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D017353:Gene Deletion; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D055542:alpha-Globins / Q000235:genetics*
chemical_list: D055542:alpha-Globins