PALM结合低体积扩增技术在混合斑检验中的应用

目的PALM系统联合应用低体积扩增技术,建立一种提取混合斑中精子DNA的分型方法。方法利用PALM切割捕获理想检材的精子细胞,将捕获的细胞直接放置到低体积扩增玻片的反应位点上,裂解细胞之后进行PCR扩增。结果20个精子细胞8次分型中,7次分型结果完全正确。10个精子细胞8次分型中,4次分型结果完全正确。1个精子细胞8次分型中,出现位点的基因座均在10个以上。将该方法应用于两例实际案例的混合斑检材,取得了满意效果。结论联合运用PALM系统和低体积扩增技术对混合斑检材中的精子细胞检验具有重要意义。

The Combination of PALM with LV-PCR System and Its Application to Mixture Sample Detection

Objective PALM was combined with the AmpliGrid LV-PCR system to establish a set of sperm DNA genotyping method from mixture sample. Methods Sperm cells were captured by PALM and placed on AmpliGrid reaction position. After incubation with PK and DTT, PCR was conducted. Results When twenty sperm cells were detected for eight repeats, seven times they were completely genotyped. Ten sperm cells were de-tected for eight repeats, four times they were completely genotyped. After eight repeat experiments on a single sperm cell assay, more than ten loci were genotyped. Finally, two mixture samples of real cases were successfully typed by this method. Conclusion The combination of PALM with LV-PCR system to resolve mixed sperm sam-ples is of great significance.