| 摘要: |
| ［摘要］ 目的 分析两种不同方法提取人粪便进行DNA检测的效果。方法 收集108例研究者的粪便，采用粪便DNA提取试剂盒及酚-氯仿法提取DNA，比较两种方法提取DNA的效果。结果 采用酚-氯仿的方法及试剂盒提取的DNA浓度分别为（232.51±41.24）ng/μl、（67.35±13.17）ng/μl（P<0.01）；试剂盒提取粪便DNA在纯度(OD260/280)、溶液澄清度及电泳结果等方面，均高（优）于酚-氯仿提取的DNA（P<0.01）。酚-氯仿及试剂盒首次提取的DNA能进行PCR扩增的阳性率分别为5%（1/20）和91.67%（99/108）（P<0.01）。另有9例采用试剂盒未能提取足够的DNA量，加大粪便量后，再次提取的DNA能够满足实验要求。结论 试剂盒提取的DNA质量较高，能满足DNA甲基化检测的条件。酚-氯仿的提取方法有待进一步研究才可能获得质量较高的DNA。 |
| 关键词: 结直肠癌 早期诊断 粪便DNA提取 |
| DOI：10.3969/j.issn.1674-3806.2013.05.03 |
| 分类号:R 735 |
| 基金项目:广西教育厅科研课题（编号：201012MS043）；广西卫生厅科研课题（编号：Z2012246） |
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| Research on the effectiveness of two methods for fecal DNA extraction and detection |
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HE Chun-gang, Huang Qin-yuan,Chen Li-sheng, et al.
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Department of General and Pediatric Surgery，the People’s Hospital of Guangxi Zhuang Autonomous Region,Nanning 530021,China
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| Abstract: |
| ［Abstract］ Objective To analyze the effectiveness of two methods for fecal DNA extraction and detection.Methods In 108 patients who were investigated，fecal samples were collected. The fecal DNA was extracted by QIAamp DNA Stool Mini Kit and phenol/chloroform method.The DNA quality extracted by the two methods was compared with each other.Results The concentrations of DNA extracted by phenol/chloroform method and kit were(232.51±41.24)ng/μl and (67.35±13.17)ng/μl, respectively (P<0.01). The fecal DNA extracted by kit outweighed than phenol/ chloroform method when compared on the indexes of purity(OD260/280)，clarity of DNA solution and electrophoresis results（all P<0.01）. There were 91.67%（99/108）samples of fecal DNA extracted by kit can be amplified by PCR, while only 5%（1/20）samples of fecal DNA extracted by phenol/chloroform method can be amplified（P<0.01）. There was not enough concentration of DNA got from another 8.33%（9/108）samples of stool at the first time extracted by kit, but can achieve the requirements by improving the method at last.Conclusion The fecal DNA may be got higher quality extracted by kit when compared with phenol/chloroform method, and can achieve the requirements of bisulfite modification and PCR.Improving the phenol/chloroform method may also get high qualitative DNA. |
| Key words: Colorectal cancer Early diagnosis Fecal DNA extraction |
