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研究论文

Detection of indicator bacteria in water bodies using polymerase chain reaction

摘要：对聚合酶链式反应(polymerasechainreaction,PCR)快速检测水体指示菌---大肠菌群(totalcoliform)和大肠杆菌(Escherichiacoli)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150bpDNA片段。引物Ⅰ和引物Ⅱ最低能分别扩增10^-6μg或10^-8μg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,是有效监测环境水体指示菌的一种潜在的新方法.

Abstract：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 260bp and 150bp respectively.In serial dilution experiments,as few as 10-100 bacteria and 10^-6μg genomic DNA were detected by primer for LacZ gene in total coloform and 1-10 bacteria and 10^-8μg 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.