中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (11): 2015-2018.doi: 10.3969/j.issn.1673-8225.2012.11.027

• 组织构建基础实验 basic experiments in tissue construction • 上一篇    下一篇

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*★

宋美茹1,姚  萍1,张跃新2   

  1. 新疆医科大学第一附属医院,1消化科,2感染科,新疆维吾尔自治区乌鲁木齐市 830054
  • 收稿日期:2011-12-08 修回日期:2012-01-29 出版日期:2012-03-11
  • 通讯作者: 姚萍,硕士,主任医师,硕士生导师,新疆医科大学第一附属医院消化科 pingyaozh@sina.com
  • 作者简介:宋美茹★,女,汉族,1978年生,陕西省人,主治医师,新疆医科大学在读硕士,主要从事溃疡性结肠炎与肠道菌群的研究。 smr771105@sina.com
  • 基金资助:

    新疆维吾尔自治区重点学科(内科学)建设基金(2004-2011)。

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

Song Mei-ru1, Yao Ping1, Zhang Yue-xin2   

  1. 1Department of Gastroenterology, 2Department of Infectious Disease, First Affiliated Hospital of Xinjiang Medical University, Urumqi  830054, Xinjiang Uygur Autonomous Region, China
  • Received:2011-12-08 Revised:2012-01-29 Online:2012-03-11
  • Contact: author: Yao Ping, Master, Chief physician, Department of Gastroenterology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China pingyaozh@sina.com
  • About author:Song Mei-ru★, Studying for master’s degree, Attending physician, Department of Gastroenterology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China smr771105@sina.com   
  • Supported by:

    Science Key Subject Construction Foundation of Xinjiang Uygur Autonomous Region, No. 2004-2011*

摘要:

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A260/280 nm值及提取率,设计乳酸菌属16S rDNA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。

关键词: 基因组DNA, QIAamp试剂盒, Biomed试剂盒, 粪便细菌, 乳酸菌属, 实时荧光定量聚合酶链反应, 组织工程

Abstract:

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.
 

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