Taqman—MGB荧光定量PCR测定饮用水中大肠菌群

目的建立饮用水中大肠菌群荧光定量PCR定量检测方法。方法以大肠菌群lacZ基因为靶基因，建立Taqman—MGB探针荧光定量PCR定量方法测定饮用水中大肠菌群，并进行方法学评价。结果25山荧光定量PCR反应体系中Mg3＋为5．0mmol／L，dNTPs0．2mmol／L，引物0．2μmol／L，探针0．5μmol／L，TaqDNA聚合酶2．0U，加入样品模板5μl。检测方法的特异性、重复性好，灵敏度高，可检出单个拷贝大肠菌群模板。结论所建立荧光定量PCR方法可快速、灵敏、特异检测饮用水中大肠菌群。

Detection of Escherichia coli from drinking water by using Taqman-MGB fuorescence quantitative PCR method

Aim To establish specific and sensitive afluorescenee quantitative PCR assay for rapidly detection of eoliforms in drinking water. Methods The target gene of lacZ was obtained and the primers and Taqman-MGB probe for PCR amplification were designed and PCR reaction system was established for detection of colfforms in drinking water. The sensitivity,specificity and reproducibility of the invcsrtiagtion were evaluated. Results The fluorescence quantitative PCR method was well established. The best 25μl reaction components was 5.0mmol/L Mg2＋ ,0.2mmol/L dNTPs,0.2 μmol/L primers,0.5 μmol/L probe,4xROX Reference Dye 0.5μl,2.0U TaqDNase and 5μL iemplate. The detection limit for coliforms was single copies in each microlitre of coliform template. The method had the advantages ors good sensitivity,specificity and reproducibility. Conduslons The fluorescence quantitative PCR method can rapidly and exactly detect the coliforms in drinking water.