应用两种PCR法快速检测公共场所水环境中嗜肺军团菌

Rapid detection of legionella pneumophila in water environment of public places by two PCR methods

目的应用巢式PCR和荧光定量PCR法调查公共场所水环境中嗜肺军团菌的污染现状与水平。方法于2012年8月-2012年11月在我国4个地区的公共场所中采集冷却水、自来水、淋浴水和喷泉水,巢式PCR检测嗜肺军团菌的污染现况,荧光定量PCR检测其污染水平。结果巢式PCR和荧光定量PCR检测公共场所水环境中嗜肺军团菌的阳性率分别为51.7%(90/174)和47.1%(82/174),两种方法的检出率差异具有统计学意义(χ2=98.083,P=0.000)。4个地区公共场所水环境嗜肺军团菌浓度均值为1.08×107copies/ml。结论巢式PCR和荧光定量PCR可作为公共场所水环境嗜肺军团菌快速检测方法之一。冷却水和淋浴水中嗜肺军团菌污染水平较自来水和景观水严重。

Objective To investigate the current contamination status and level of legionella pneumophila in water environment of public places by nested PCR and fluorescent quantitative PCR. Methods The cooling water, tap water, shower water and fountain water of public buildings were collected from 4 areas of China from August to November in 2012. The current pollution status of legioneUa pneumophila was detected in water samples by nested - PCR, while the pollution level of that was detected by real - time PCR. Results The positive rate of legioneUa pneumophila in water environment of public places was 51.7% （90/ 174） by nested - PCR and 47.1% （82/174） by real - time PCR, and the difference had statistical significance （X2 = 98. 083, P = 0.000）. The mean concentration of legionella pneumophila in water environment of public places of the 4 areas was 1.08 ×10^7 copies/ml. Conclusion Nested - PCR and real - time PCR can be applied to rapid detection of the legionella pneumophila in the water environment of public places. The contamination levels of cooling water and shower water are higher than the tap water and fountain water.