• 关键词：通用引物  病原细菌  定量PCR  膜过滤  地表水

• 基金项目：国家自然科学基金项目(No50478048);国家自然科学基金重大国际合作项目(No50621140001);西安市科技计划项目(NoYF07196)

• 刘永军
• 西北水资源生态与环境教育部重点实验室, 西安建筑科技大学, 西安 710055

• 张崇淼
• 西北水资源生态与环境教育部重点实验室, 西安建筑科技大学, 西安 710055

• 王晓昌
• 西北水资源生态与环境教育部重点实验室, 西安建筑科技大学, 西安 710055

• 吕英俊
• 西北水资源生态与环境教育部重点实验室, 西安建筑科技大学, 西安 710055

• 李昂臻
• 西北水资源生态与环境教育部重点实验室, 西安建筑科技大学, 西安 710055

• 薛小平
• 西北工业大学, 生命科学院, 西安 710072

• 摘要：采用通用引物,通过实时荧光定量PCR方法(QPCR),对西安市5个地表水体中肠道病原细菌的细胞密度进行了分析.检测水样体积为100mL,分别于2006年3～6月取自水源水(黑河)、景观娱乐用水(大唐芙蓉园北湖和兴庆湖)、纳污河(浐河)和未消毒的二级出水(北石桥污水处理厂),并将QPCR检测结果与滤膜法(MF)测得的细菌总数、大肠菌群和粪大肠杆菌CFU结果进行了比较分析.5个水体(n=60)检测结果显示,QPCR检测结果是大肠菌群CFU的2.2～5倍,是粪大肠杆菌CFU的7～14倍.病原细菌检测的几何平均值范围,QPCR法在25～67000CCE·100mL^-1之间,MF法大肠菌群在3～45000CFU·100mL^-1之间,粪大肠菌群在0～3000CFU·100mL^-1之间(n=60).两种方法的检测结果使用Spearman秩相关法来计算,结果显示,QPCR检测结果与大肠菌群、细菌总数以及粪大肠杆菌CFU均呈现显著正相关,秩相关系数分别为r=0.983、r=0.908和r=0.948.

• Abstract：Arapid Quantitative Polymerase Chain Reaction (QPCR) analysis method with universal primers was developed to detect cell densities of enteric pathogenic bacteria in surface water. Water samples with a volume of 100 mLwere collected from five surface waters of Xi′an including a source water for the drinking water supply (Heihe River), a newly built artificial lake (Beihu Lake), a polluted lake after water quality restoration (Xingqinghu Lake), a water course receiving industrial discharge (Chanhe River) and a secondary effluent without disinfection (Beishiqiao Wastewater Treatment Plant) during March to June 2006 (3 times every month). Measurements by the QPCRmethod were compared with counts of total bacteria, total coliforms, and fecal coliforms colony-forming units (CFU) determined by membrane filter (MF) analysis. The geometric mean of pathogenic bacteria concentration determined by QPCR from the five surface waters(n=60)was 2.2～5 times the coliform CFUand 7～14 times the fecal coliforms CFUas determined by MF analysis. Geometric means of pathogenic bacteria concentration determined by QPCRranged from 25～67000CCE·100mL^-1. Total coliforms and fecal coliforms by the MF method ranged from 3～45000 CFU·100mL^-1 and 0～3000 CFU·100mL^-1, respectively(n=60). Spearman's analysis showed that there was a significant positive correlation between pathogenic bacteria determined by QPCR and total bacteria, coliforms, and fecal coliforms determined by MF. The correlation coefficients (r) were 0.983, 0.908 and 0.948 for total bacteria, total coliforms, and fecal coliforms, respectively.

• Key words：universal primer  enteric pathogenic bacteria  quantitative polymerase chain reaction(QPCR)  membrane filtration (MF)  surface water

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