城市污水二级处理出水中沙门氏菌的PCR检测  

PCR Detection of Salmonella in Secondary Effluent in Municipal Wastewater Treatment System

作　　者：曾颂[1] 张崇淼[1] 王晓昌[1] 魏梦楠[1] 

机构地区：[1]西安建筑科技大学,西北水资源与环境生态教育部重点实验室,陕西西安710055

出　　处：《环境科学研究》2010年第3期361-365,共5页Research of Environmental Sciences

基　　金：国家自然科学基金重点项目(50838005);西安建筑科技大学基础研究基金(JC0910)

摘　　要：建立了一种利用PCR检测城市污水二级处理出水中沙门氏菌的方法.采用膜吸附-洗脱法浓缩二级出水水样中的沙门氏菌,通过选用一对涵盖性和特异性均较强的引物139和141,对浓缩后的沙门氏菌进行PCR检测.利用正交试验法,对沙门氏菌PCR体系进行五因素四水平的试验,建立了沙门氏菌PCR的最佳体系.通过梯度PCR考察了退火温度对检测结果的影响,选择65℃作为退火温度,优化后沙门氏菌PCR体系的检测灵敏度可达0.314ngL.通过对已知细菌浓度的水样进行检测,确定了该法检测水样中沙门氏菌的灵敏度为2130CFUL.与传统MPN法的相比,该法在检测效果和检测效率上都优于MPN法.A method was established for the detection of Salmonella from secondary effluent in municipal wastewater treatment system with the application of PCR technique.The water sample was concentrated by membrane adsorption-elution operation,and then the concentrated Salmonella was detected using a pair of primers 139 and 141,which have both higher generality and specificity.The optimal condition for the PCR detection was obtained by orthogonal experiments regarding five factors at four levels.As a result,65℃ was determined as the optimal annealing temperature by gradient PCR,and the detection limit of the optimized PCR was 0.314 ng/L.Through a series of measurements using spiked water samples,the detection limit was further found to be 2130 CFU/L if the method was applied to practical water samples.The method established in this study showed its advantage over the conventional MPN method in its higher sensitivity and efficiency.

关 键 词：二级处理出水 沙门氏菌 浓缩 正交设计 优化 聚合酶链式反应(PCR) 

分 类 号：R123[医药卫生—环境卫生学] X832[医药卫生—公共卫生与预防医学]