三种定量方法在温泉水中军团菌连续性监测中的应用  

Three quantitative methods to continuously monitor Legionella in spring water

作　　者：闫革彬[1] 王焕新[1] 秦天[2] 周海健[2] 李马超[2] 徐颖[1] 赵明强[1] 邵祝军[2] 任红宇[2] 

机构地区：[1]北京市昌平区疾病预防控制中心传染病防治科,102200 [2]中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室

出　　处：《中华预防医学杂志》2013年第7期637-640,共4页Chinese Journal of Preventive Medicine

基　　金：中国疾病预防控制中心青年基金（2011A102）;传染病预防控制国家重点实验室面上项目（2011SKLID202）

摘　　要：目的比较传统的平板培养法、荧光定量PCR和荧光染料叠氮溴化乙锭（ethidiummonoazide，EMA）结合荧光定量PCR的方法（EMA-荧光定量PCR）对温泉水中军团菌的检测效果。方法于2011年每月（除5月份）对北京市某温泉度假村的5个温泉池进行采样，每月采集11份水样，全年共采集121份。分别采用传统的平板培养法、荧光定量PCR和EMA-荧光定量PCR方法对水样中军团菌进行定性和定量分析。结果平板培养法、荧光定量PCR和EMA-荧光定量PCR检测温泉水中军团菌污染率分别为74．4％（90／121）、100．0％（121／121）和100．0％（121／121）；3种方法对121份温泉水样本中军团菌定量数值分别为0．10-216．00菌落形成单位（CFU）／ml、1．47-1557．75基因单位（GU）／ml和0．20-301．69GU／ml。荧光定量PCR、EMA-荧光定量PER和平板培养方法对121份温泉水每份军团菌含量检测的中位数（第25百分位数～第75百分位数）分别为75．30（32．51-192．10）GU／ml、36．46（16．08-91．21）GU／ml和5．30（0．00～33．70）CFU／ml。对于121份温泉水水样，3种方法对军团菌含量分析的定量数值差异有统计学意义（X^2=187．900，P〈0．01），其中荧光定量PCR的定量数值最高，EMA-荧光定量PCR的数值次之，平板培养的数值最低。结论荧光定量PCR和EMA-荧光定量PCR方法检测温泉水中军团菌的灵敏度优于传统的培养方法，EMA-荧光定量PCR方法适合用于环境水体中军团菌活菌监测。Objective To compare the detection effect of LegioneUa pollution in spring water by three methods, namely traditional plating method, fluorescent quantitation PCR method and ethidium monoazide （EMA） fluorescent quantitation PCR method. Methods Every month （except May）, we collected 11 water samples from the 5 selected hot spring pools in one hot spring resort in Beijing in 2011. A total of 121 water samples were collected, and then were detected by the above three methods qualitatively and quantitatively. Results In our study, the Legionella pollution rate was separately 74.4% （90/121）, 100. 0% （121/121） and 100. 0% （121/121） by the above three methods. The quantitative value of Legionella in the 121 water samples detected by the three methods were around 0. 10 -216.00 colony-forming units （ CFU）/ml, 1.47 - 1557.75 gene units （ GU）/ml and 0. 20 - 301.69 GU/ml, respectively. The median （ 25th and 75th percentiles） was 75.30 （ 32. 51 - 192. 10） GU/ml, 36.46 （ 16. 08 -91.21 ） GU/ml and 5.30 （0. 00 -33.70） CFU/ml,respectively. The difference in the quantitative value of Legionella detected by the three methods showed statistical significance （X^2 = 187. 900, P 〈 0.01 ）. The quantitative value of Legionella detected by fluorescent quantitation PCR method was the highest, followed by the value Legionella detected by EMA-fluorescent quantitation PCR method and traditional platingmethod. Conclusion The sensitivity of the PCR methods was higher than traditional plating method, in detecting Legionella pollution in spring water, especially the EMA- fluorescent quantitation PCR method, which was more suitable for detecting Legionella in water.

关 键 词：军团菌 长滩 温泉 聚合酶链反应 荧光染料 

分 类 号：R[医药卫生]