定量PCR结合STR多态连锁分析诊断缺失型DMD基因携带者

Detection of DMD carriers in deletion families by quantitative polymerase chain reaction and the linkage analysis of short tandem repeat polymorphisms.

目的 直接定量PCR结合STR单体型连锁分析 ,以便更准确检出缺失型家系中DMD基因携带者。方法 在内标引物参照下 ,PCR扩增 2 2个循环 ,产物在琼脂糖凝胶分离 ,EB显色 ,照相后 ,扫描定量 ,同时应用 5个STR多态位点的单体型连琐分析 ,检测缺失型的DMD基因女性携带者。结果 直接定量PCR分析了 6个家系中的 8个外显子 48缺失的DMD母亲或女性同胞 ,检出 7个DMD为携带者 ;STR多态单体型连锁同样分析这 6个家系中的 8个女性亲属 ,亦检出 7个DMD基因携带者 ,二种方法所得结果完全一致。结论 STR多态连锁分析与直接定量PCR法结合 ,可更准确 ,全面检出DMD基因携带者。

Objective:To detect DMD gene carriers in the deletion families accurately,using quantitative PCR technique and the linkage analysis of STR polymorphisms.Methods: DNA samples are abstracted from females in exon 48 deletion DMD families.PCR are performed for 22 cycles,contrasted with internal standard primer located in this gene.After separated on 1.5% agaros gel electrophoresis and stained with EB the separated bandings of PCR products are taken photography and performed quantitative analysis on TLC densitometer.At the same time,the monosomy linkage analysis are performed with 5 loci of STR polymorphisms.Results: 8 cases of possible carriers in 6 DMD families with exon 48 locus deletion are detected by quantitative PCR and 7 carries with deletion DMD gene are diagnosed. And 7 cases of DMD gene Carriers in 6 same deletion DMD families are detedted using the linkage analysis of STR polymorphisms.The results are identical between two analysis methods above.Conclusion: Using two methods of the linkage analysis of STR polymorphisms and quantitative PCR technique,can be detected the carriers in DMD deletion families more accurately and more effectively.