南方医科大学学报2017，Vol.37Issue(9)：1265-1269,5.DOI:10.3969/j.issn.1673-4254.2017.09.22
微滴式数字PCR(ddPCR)快速检测α珠蛋白基因αααanti-3.7三联体
Rapid detection of alpha-globin gene αααanti-3.7 triplets with droplet digital PCR
摘要
Abstract
Objective To establish a rapid method for detection of alpha-globin gene αααanti-3.7 based on droplet digital PCR (ddPCR) technique.Methods The differential sequence between the X1 and Y1 box of α1 gene was selected as the amplicon of the target gene with β-actin as the reference gene.The specific primers and TaqMan probes were designed,and then a quantitative method for detecting the copy number was established based on ddPCR technique.The sensitivity and accuracy of the method were evaluated by detecting 28 samples of known genotypes and 60 clinical samples.Results The ddPCR-based method accurately identified the genotypes of all the 28 samples with known genotypes and detected 5 cases of αα/αααanti-3.7 from the 60 clinical samples,and the results were verified by MLPA.The sensitivity and accuracy of this method were both 100％ for detecting alpha-globin gene αααanti-3.7.Conclusion This ddPCR-based method for detecting αααanti-3.7 triplet can be applied for population screening and in routine clinical molecular diagnosis with simple operation,rapid analysis and accurate results.关键词
数字PCR/珠蛋白基因/多拷贝/地中海贫血Key words
digital polymerase chain reaction/globin gene/multi-copy/Thalassemia引用本文复制引用
龚小倩,杨学煌,乔伶俐,陈亚军,周万军..微滴式数字PCR(ddPCR)快速检测α珠蛋白基因αααanti-3.7三联体[J].南方医科大学学报,2017,37(9):1265-1269,5.基金项目
韶关市科技计划项目(2015CX/K19) （2015CX/K19）
