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目的探讨多重置换扩增(MDA)技术对法医学微量DNA样品STR检测分型的效果。方法用MDA技术对不同模板量DNA进行全基因组扩增(WGA),扩增产物用实时荧光定量PCR技术定量、用Profiler PlusTM试剂盒检测基因型。结果该方法可对模板DNA增加104～106倍。1ng样品DNA的MDA产物可获得9个STR基因座和Amelogenin性别基因座的准确分型结果;低于0.1ng的样品DNA经MDA扩增后,基因座检出数增加,但可见等位基因不平衡或丢失现象。结论MDA技术可有效增加DNA模板量和提高微量DNA分型效果。但样品DNA量低于0.1ng时,MDA产物的STR分型结果判读须慎重。
Objective To investigate the effect of multiple replacement amplification (MDA) on the detection and typing of microsatellite DNA in forensic medicine. Methods Genome-wide amplification (WGA) was performed on different template DNA using MDA. The amplified products were quantified by real-time fluorescence quantitative PCR and the genotypes were detected by Profiler PlusTM kit. The results of this method can increase the template DNA 104 to 106 times. 1ng sample DNA MDA product obtained 9 STR loci and Amelogenin gender loci accurate typing results; less than 0.1ng sample DNA amplified by MDA, the number of loci detected increased, but we can see the allele is not Balanced or lost. Conclusion MDA can effectively increase the amount of DNA template and improve the DNA typing efficiency. However, when the sample DNA amount is less than 0.1ng, the STR typing results of the MDA product should be interpreted judiciously.