QIAamp及Biomed粪便细菌基因组DNA提取试剂盒提取人肠道细菌基因组DNA质量的差异

背景:有研究表明,不同试剂盒提取的粪便细菌基因组DNA质量有差别,选择一种操作简便、质量优良的粪便细菌基因组DNA提取试剂盒成为研究者们关注和急需解决的问题。目的:比较QIAamp及Biomed粪便细菌基因组DNA提取试剂盒提取的人肠道细菌基因组DNA质量的差异。方法:收集健康成年人新鲜粪便标本30例,用两种试剂盒分别提取细菌基因组DNA,检测其浓度、吸光度A260/280nm值及提取率,设计乳酸菌属16SrDNA基因特异性引物,以各自所提DNA为模板,进行常规聚合酶链反应,凝胶电泳后比较条带数量、明暗度及密度,并进行实时荧光定量聚合酶链反应定量检测各自所含乳酸菌属的数量。结果与结论:QIAamp试剂盒提取的正常人肠道细菌基因组DNA的浓度、提取率、表达量及乳酸菌属的数量均高于Biomed试剂盒(P<0.05或P<0.01)。说明QIAamp试剂盒提取的粪便细菌基因组DNA质量优于Biomed试剂盒。

Quality differences of bacterial genome DNA extracted from the human intestine by using QIAamp and Biomed DNA Stool Mini Kits

BACKGROUND: Several studies have showed that there are different qualities of bacterial genome DNA extracted from the human intestine using different kits. Therefore, it is essential and urgent to select an effective, simple, and excellent DNA stool mini kit. OBJECTIVE: To compare the quality differences of bacterial genome DNA extracted from the human intestinal by using QIAamp and Biomed DNA Stool Mini Kits. METHODS: Thirty fresh fecal samples of healthy adult people were selected, and bacterial genome DNA was extracted using these two kits. Concentration, absorbance radio of 260 to 280 nm and extraction rate were measured. Species-specific primers for Lactobacillus group were designed by a set of 16S rDNA-targeted to conduct the general PCR using the genome DNA as the template. The electrophoresis strip’s number, brightness and density were compared after gel electrophoresis; subsequently, the number of Lactobacillus group was detected by fluorescent real-time PCR. RESULTS AND CONCLUSION: Concentration, extraction rate, expression level , Lactobacillus amount of bacterial genome DNA extracted from the human intestinal using QIAamp kit were higher than those using Biomed kit (P < 0.05 or P < 0.01). It suggests that the quality of genome DNA extracted with QIAamp kit is more excellent than that with Biomed kit.