摘要
对聚合酶链式反应（ｐｏｌｙｍｅｒａｓｅ ｃｈａｉｎ ｒｅａｃｔｉｏｎ，ＰＣＲ） 快速检测水体指示菌———大肠菌群（ｔｏｔａｌ ｃｏｌｉｆｏｒｍ） 和大肠杆菌（ Ｅｓｃｈｅｒｉｃｈｉａ ｃｏｌｉ） 进行了研究．采用的两对引物分别扩增了ＬａｃＺ基因的约２６０ｂｐ 和ＵｄｉＡ 基因的约１５０ｂｐ ＤＮＡ 片段．引物Ⅰ和引物Ⅱ最低能分别扩增１０ － ６μｇ 或１０ －８ μｇ 基因组ＤＮＡ．整个检测工作在采集水样后５ —６ｈ 内完成．与常规水质检测法相比，ＰＣＲ 法具有快速，准确的特点。
Polymerase chain reaction (PCR) procedure was applied for identification of two indicators,total coliform and E.coli ,in various water samples.Two oligonucleotide primers were used in this procedure to amplify LacZ gene DNA fragments of total coloform and UdiA gene DNA fragments of E.coli respectively.According to the results of agarose gel electrophoresis,the length of the DNA fragments amplified were about 260?bp and 150?bp respectively.In serial dilution experiments,as few as 10—100 bacteria and 10\+\{-6\}?μg genomic DNA were detected by primer for LacZ gene in total coloform and 1—10 bacteria and 10\+\{-8\}?μg genomic DNA by primer for UdiA gene in E.coli .Only 5—6 hours were needed to finish the assay after samples gathered.A total 40 water samples were detected by the PCR procedure and multi\|tube\|fermentation(MTF) method.The coincidence of the two methods was 100% in big volume water sample and 77 5% in small volume water sampel.
出处
《环境科学学报》
CAS
CSCD
北大核心
1999年第5期540-544,共5页
Acta Scientiae Circumstantiae
基金
浙江省科委项目
浙江省教委项目
