rep-PCR基因指纹图微生物源追踪方法建立的研究  

STUDY ON THE REPETITIVE EXTRAGENIC PALINDROMIC PCR FINGERPRINT ANALYSIS AS A MICROBIAL SOURCE TRACKING

作　　者：李伟伟[1] 顾玲 叶珣[3] 陈晓东[1,2] 丁震 周璐 董晨 张琪[1] 于洋[1] 汪华 

机构地区：[1]南京医科大学公共卫生学院,南京210029 [2]江苏疾病预防控制中心 [3]东南大学公共卫生学院

出　　处：《现代预防医学》2009年第17期3242-3244,共3页Modern Preventive Medicine

基　　金：江苏省"333工程"培养资金资助项目

摘　　要：[目的]建立重复序列聚合酶链反应(rep-PCR)微生物源追踪方法,应用于淮河流域某水库的粪便污染来源追踪。[方法]采集已知来源的人和动物粪便标本以及水样标本,分离并确认指示菌大肠杆菌,以BOXA1R为引物进行PCR扩增;用Bionumerics 4.0软件对rep-PCR基因指纹图进行判别分析和多元方差分析并计算各源种类的正确归类率。[结果]2分类、4分类和10分类的正确归类率分别为:85.60%、79.20%和70.98%。判别分析和多元方差分析结果显示rep-PCR基因指纹图能将已知源不同来源指示菌区分开。水样中主要粪便污染来源为猪、鸡和人。[结论]建立的rep-PCR微生物源追踪方法能较好地区分水体中指示菌的不同来源,该方法的建立为查找水体中粪便污染来源以及水体治理提供了新技术。[Objective] The rep-PCR fingerprinting analysis was used as a microbial source tracking method and was applied to analysis a reservoir of the Huai River. [ Methods ] Fresh fecal materials were collected from human and animals throughout villages around the reservoir .Escherichia coli were identified as indicators and PCR was expanded with the support of BOXA1R premier. The rates of correct assignment, Multivariate analysis of variance （MANOVA） and discriminant analysis were conducted by Bionumerics 4.0 software. Water Samples were applied to identify the main fecal pollutions. [ Results3 The known source fingerprint library which was classified as 2, 4 and 10 groups, the average rates of correct classification were re- spectively 85.60%, 79.20% and 70.98%. MANOVA and discriminant analalysis showed that the division of different groups was clearly. The reservoir was most possibly polluted by swine, cow and human. [ Conclusion ] The rep-PCR DNA fingerprint anal- ysis is a promising method for determining the source groups of indictor isolates in water, it can be used to track the fecal pollu- tions source and help to provide a new method for water environment treatment.

关 键 词：微生物源追踪 REP-PCR BOXAIR 大肠杆菌 

分 类 号：R123.1[医药卫生—环境卫生学]