3种DNA提取法在污染严重混合斑分型中的应用比较

发布时间：2018-10-19 09:13

【摘要】：目的比较Chelex-100法、酚/氯仿法和二氧化硅膜法3种DNA提取法在污染严重混合斑分型中的应用效果。方法从日常案例中收集污染严重的混合斑25份,差异消化法分离精子后同时用Chelex-100法、酚/氯仿法和二氧化硅膜技术3种方法提取DNA,采用PCR-STR技术对D19S253、FGA和CSF1PO 3个基因座进行分型,Gel-Pro软件处理电泳图谱,SPSS软件分析比较不同方法之间的差异。结果采用Chelex-100法提取DNA,25份检材分型结果均未成功;采用酚/氯仿法,25份检材中10份分型成功,3份检材FGA和CSF1PO基因座可分型,4份检材CSF1PO基因座可分型;采二氧化硅膜纯化法,25份检材均成功分型;酚/氯仿法和二氧化硅膜法两种方法比较,结果存在显著性差异(P0.05)。结论二氧化硅膜纯化技术可以有效去除PCR抑制物,提取的DNA扩增效果明显优于Chelex-100法和酚/氯仿法,具有较高的应用价值。
[Abstract]:Objective to compare the application effect of Chelex-100 method, phenol / chloroform method and silica membrane method in the classification of seriously polluted mixed spot. Methods Twenty-five samples of seriously polluted mixed spots were collected from daily cases. Sperm were separated by differential digestion and Chelex-100 was used at the same time. DNA, was extracted by phenol / chloroform method and silica membrane technique. Three genotypes of D19S253FGA and CSF1PO were classified by PCR-STR technique. The electrophoretic patterns were processed by Gel-Pro software and the differences between different methods were analyzed and compared by SPSS software. Results the typing results of DNA,25 samples extracted by Chelex-100 method were not successful, 10 of 25 samples were successfully typed by phenol / chloroform method, 3 samples were genotyped by FGA and CSF1PO loci, and 4 samples by CSF1PO loci. Silica membrane purification method, 25 samples were successfully classified; phenol / chloroform method and silica membrane method were compared, the results showed significant difference (P0.05). Conclusion silica membrane purification technique can effectively remove PCR inhibitor, and the amplification effect of DNA extracted is better than that of Chelex-100 and phenol / chloroform method.
【作者单位】：华中科技大学同济医学院法医学系河南科技大学法医学院华中科技大学同济医学院法医学系
【分类号】：D919

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