广西医学Issue(8)：984-988,5.DOI:10.11675/j.issn.0253-4304.2013.08.07
双重 TaqMan 荧光定量 PCR 在β-地中海贫血产前诊断中的应用▲
Dual TaqMan Fluorescence Quantitative PCR Applied to Prenatal Diagnosis of beta-Thalassemia
摘要
Abstract
Objective To establish a dual Taqman fluorescence quantitative PCR ( qPCR ) method for point mutation detection , and to realize the fast and accurate prenatal diagnosis of high-risk fetus with beta-thalassemia . Methods A dual Taqman qPCR method for beta-thalassimia point mutation detection was developed according to the design that mutation and wild type sequences of a mutation site analyzed in one reaction tube .This method was also applied to detect 6 villi and 32 amniotic fluid specimens of high-risk fetus with beta-thalassemia .The genotype of a sample was analyzed according to the positive amplification results of qPCR and Ct variance of two fluorescent chan -nels.Meanwhile,reverse dot blot(RDB) method was used as a control for the analysis and evaluation of detection re-sults.Results Of 6 villi specimens ,1 sample which could not be diagnosed with RDB method was detected wild type homozygote by this method ,and another sample with the misdiagnosis RDB result of double heterozygote of βCD41-42 and βCD26 was detected heterozygote of βCD41-42 by this method .Of 32 amniotic fluid specimens ,1 sample which could not be diagnosed with RDB method was detected heterozygote of βCD41-42 by this method .All these 3 samples were also re-analyzed and verified with recollected amniotic fluid specimens .Conclusion The fast and accurate pre-natal diagnosis of high-risk fetus with beta-thalassemia can be realized with this dual Taqman qPCR method for beta gene point mutation developed in this study .The method is accurate with easy operation and suitable for prenatal diag-nosis of beta-thalassemia .关键词
β-地中海贫血/双重TagMan荧光PCR技术/产前诊断/基因型Key words
beta-thalassemia/Dual Taqman fluorescence quantitative PCR/Prenatal diagnosis/Genotype分类
医药卫生引用本文复制引用
陈碧艳,邓建平,覃茜..双重 TaqMan 荧光定量 PCR 在β-地中海贫血产前诊断中的应用▲[J].广西医学,2013,(8):984-988,5.基金项目
广西百色市科学研究与技术开发计划项目 （）
