两种全自动核酸纯化仪的应用

Application of Two Kinds of Automatic Nucleic Acid Purifiers

目的探讨QIAsymphony SP和QIAcube全自动核酸纯化仪在微量和含抑制剂类检材中的DNA提取效果。方法应用QIAsymphony SP和QIAcube的不同纯化方法提取含有30、100、150和300个细胞制成的棉签拭子以及含有血红素、腐殖酸、猪油、土壤、铁锈和润滑油6种类型抑制剂的样本,将提取到的DNA模板进行PCR扩增和STR分型,比较不同纯化方法的提取效率和抑制剂去除能力。结果不同纯化方法的提取效果相当,30、100和300个细胞的样本提取得到的DNA,经扩增检测均有70.83%~100.00%的基因座检出,且均能很好地去除上述6种类型抑制剂。结论两种全自动核酸纯化仪均有良好的抑制剂去除效果,对于仅有30个细胞的棉签拭子,经过DNA提取扩增之后,样本也有较高的基因座检出率,能够满足基层DNA实验室工作的需求,实现实验室的标准化建设。

Objective To investigate the effect of automatic nucleic acid purifiers QIAsymphony SP and QIAcube in the DNA extraction of samples of trace amount or mixed with inhibitors.Methods Different kinds of purification methods using QIAsymphony SP and QIAcube were applied to extract swabs which contained 30,100,150 and 300 cells and other samples which contained six types of inhibitors-heme,humic acid,lard,soil,rust and grease.PCR amplification and STR typing were performed on the extracted DNA templates to compare extracting efficiency and inhibitor removal ability of four different purification methods.Results Different purification methods showed similar extraction effects,70.83%-100.00%of loci could be detected by amplification of DNA extracted from samples with 30,100 and 300 cells,and the six types of inhibitors could be removed well.Conclusion The two automatic nucleic acid purifiers have a good inhibitor removal effect.For swabs with only 30 cells,after DNA extraction and amplification,the locus detection rate of samples can still be high,which can meet the requirements of local DNA laboratory work,and realize the standardization construction of the laboratory.