应用FCM-qPCR方法定量检测水中常见病原体

以往对水体病原体的研究主要是通过监测粪大肠杆菌作为指示,然而研究表明粪大肠杆菌与水中病毒和细菌病原体呈现出较差的相关性.因此,选取水中典型病原体并对其进行定量检测是当前需要解决的技术问题.为此本研究建立了流式细胞术和定量PCR联合使用方法,用于快速获取水环境中总病毒、细菌以及几种典型病原体(大肠杆菌、军团菌、腺病毒、贾第虫和隐孢子虫等)的浓度水平,并将该方法应用到污水处理厂进出水及受纳河流上下游的病原体检测中.结果表明,该污水处理厂对总细菌和总病毒以及几种典型病原体都具有较高的去除率(93%);污水处理厂排水对受纳水体病原体浓度水平基本没有负面影响.研究为评估污水处理厂处理效果及排水对受纳水体的生态影响提供了技术支持.

Application of FCM-qPCR to Quantify the Common Water Pathogens

In the past, fecal E. coli was always regarded as the indicator organism for estimation of pathogens in water. However, a weak relation between fecal E.coli and water viruses or bacterial pathogens has been demonstrated by previous studies. Therefore, for water pathogen study, it is essential to select and quantify typical pathogens. In this study, a combination of quantitative PCR (qPCR) with flow cytometry (FCM) was established to detect the concentrations of viruses, bacteria and several typical pathogens (e.g. E. coli, Legionnella, HAdV, Giardia, Cryptosporidium) in water. The method was applied to measure the pathogen concentrations in the influent and effluent of a wastewater treatment plant (WWTP), as well as its receiving river. The results revealed that the WWTP treated the pathogens with high removal efficiency ( > 93%); the effluent of WWTP did not show a negative effect on pathogen concentration of the receiving river. The study provides a technical support for the evaluation of WWTP treatment effect and the ecological impact of WWTP effluent on receiving river.