粪便DNA亚硫酸盐快速处理方法的探讨  

The method research of rapid fecal DNA sulfite treatment

作　　者：孙冬生[1] 王刚[1] 李东坤[2] 张青松[2] 王志刚[3] 

机构地区：[1]唐山市开滦总医院肿瘤科,河北唐山063000 [2]唐山市开滦总医院普外科,河北唐山063000 [3]内蒙古大学生命科学学院,内蒙古呼和浩特010021

出　　处：《临床军医杂志》2015年第3期297-299,共3页Clinical Journal of Medical Officers

基　　金：中国煤炭工业协会2012年度科学技术研究指导性计划项目(MTKJ 2012-420)

摘　　要：目的探讨从微量的粪便中运用亚硫酸盐法成功处理DNA甲基化的方法,寻求最适宜的亚硫酸盐的浓度以及处理时间。方法收集我科员工新鲜粪便标本(经胃镜及结肠镜证实无消化道肿瘤),经亚硫酸盐方法处理后,对大肠埃希菌进行PCR检测,检测结果使用ABI310测序仪器进行测序。结果在最终浓度为2.63 mol亚硫酸盐、并经过95℃15min,4℃10 min,75℃60 min处理;2.11 mol亚硫酸盐经过95℃15 min,4℃10 min,75℃165 min处理均能将E.coli DNA的胞嘧啶(C)转变为尿嘧啶(U)。结论为从粪便中检测人类相关基因甲基化提供了快捷、科学的依据。Objective To explore the rapid fecal DNA sulfite treatment so as to seek for suitable concentration of sulfite and time of treatment. Methods Fresh specimens of feces were collected from our staffs without gastrointestinal tumor,and underwent PCR to detect E. coli DNA after sulfite treatment. The test results were sequenced via ABI310 Sequencer. Results At the final concentration of 2. 63 mol sulfite and after the processing of 95 ° C-15 min,4 ° C-10 min and 75 ° C-60 min,and at that of 2. 11 mol sulfite and after the processing of 95 ° C-15 min,4 ° C-10 min and 75 ° C-165 min,the cytosine of E. coli DNA was transferred into uracil. Conclusion The method provides a quick,scientific basis for the detection of human genes methylation from feces.

关 键 词：亚硫酸盐处理法 粪便DNA 基因 甲基化 

分 类 号：R446.13[医药卫生—诊断学]