title: A melting curve analysis--based PCR assay for one-step genotyping of β-thalassemia mutations a multicenter validation.
abstract: The increasing number of disease-causing mutations demands a simple, direct, and cost-effective diagnostic genotyping technique capable of detecting multiple mutations. This study validated the efficacy of a novel melting curve analysis-based genotyping assay (MeltPro HBB assay) for 24 β-thalassemia mutations in the Chinese population. The diagnostic potential of this assay was evaluated in 1022 pretyped genomic DNA samples, including 909 clinical cases of β-thalassemia minor or major, using a double-blind analysis in a multicenter validation study. Reproducibility of the assay was 100%, and the limit of detection was 10 pg per reaction. All 24 β-thalassemia mutations were accurately genotyped, and β-thalassemia genotypes were correctly determined in all 1022 samples, yielding overall sensitivity and specificity of 100%. The concordance rate was 99.4% between this assay and the reference method. It was concluded that the MeltPro HBB assay is useful for reliable genotyping of multiple β-thalassemia mutations in clinical settings and may have potential as a versatile method for rapid genotyping of known mutations because of its high throughput, accuracy, ease of use, and low cost.
authors.affiliation: Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
authors.forename: Fu
authors.initials: F
authors.lastname: Xiong
authors.forename: Qiuying
authors.initials: Q
authors.lastname: Huang
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Chen
authors.forename: Yuqiu
authors.initials: Y
authors.lastname: Zhou
authors.forename: Xinhua
authors.initials: X
authors.lastname: Zhang
authors.forename: Ren
authors.initials: R
authors.lastname: Cai
authors.forename: Yajun
authors.initials: Y
authors.lastname: Chen
authors.forename: Jiansheng
authors.initials: J
authors.lastname: Xie
authors.forename: Shanwei
authors.initials: S
authors.lastname: Feng
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Wei
authors.forename: Qizhi
authors.initials: Q
authors.lastname: Xiao
authors.forename: Tianlang
authors.initials: T
authors.lastname: Zhang
authors.forename: Shiqiang
authors.initials: S
authors.lastname: Luo
authors.forename: Xuehuang
authors.initials: X
authors.lastname: Yang
authors.forename: Ying
authors.initials: Y
authors.lastname: Hao
authors.forename: Yanxia
authors.initials: Y
authors.lastname: Qu
authors.forename: Qingge
authors.initials: Q
authors.lastname: Li
authors.forename: Xiangmin
authors.initials: X
authors.lastname: Xu
mesh_terms: D001609:Beta-Globulins / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D004311:Double-Blind Method; D006801:Humans; D057230:Limit of Detection; D025202:Molecular Diagnostic Techniques; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D044366:Transition Temperature; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics
chemical_list: D001609:Beta-Globulins