中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2013年
7期
637-640
,共4页
闫革彬%王焕新%秦天%周海健%李马超%徐颖%赵明强%邵祝军%任红宇
閆革彬%王煥新%秦天%週海健%李馬超%徐穎%趙明彊%邵祝軍%任紅宇
염혁빈%왕환신%진천%주해건%리마초%서영%조명강%소축군%임홍우
军团菌,长滩%温泉%聚合酶链反应%荧光染料
軍糰菌,長灘%溫泉%聚閤酶鏈反應%熒光染料
군단균,장탄%온천%취합매련반응%형광염료
Legionella,long beach%Hot springs%Polymerase chain reaction%Fluorochrome
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(ethidium monoazide,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0.20～301.69 GU/ml.荧光定量PCR、EMA-荧光定量PCR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(x2=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.
目的 比較傳統的平闆培養法、熒光定量PCR和熒光染料疊氮溴化乙錠(ethidium monoazide,EMA)結閤熒光定量PCR的方法(EMA-熒光定量PCR)對溫泉水中軍糰菌的檢測效果.方法 于2011年每月(除5月份)對北京市某溫泉度假村的5箇溫泉池進行採樣,每月採集11份水樣,全年共採集121份.分彆採用傳統的平闆培養法、熒光定量PCR和EMA-熒光定量PCR方法對水樣中軍糰菌進行定性和定量分析.結果 平闆培養法、熒光定量PCR和EMA-熒光定量PCR檢測溫泉水中軍糰菌汙染率分彆為74.4％ (90/121)、100.0％(121/121)和100.0％(121/121)；3種方法對121份溫泉水樣本中軍糰菌定量數值分彆為0.10 ～ 216.00菌落形成單位(CFU)/ml、1.47～1557.75基因單位(GU)/ml和0.20～301.69 GU/ml.熒光定量PCR、EMA-熒光定量PCR和平闆培養方法對121份溫泉水每份軍糰菌含量檢測的中位數(第25百分位數～第75百分位數)分彆為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.對于121份溫泉水水樣,3種方法對軍糰菌含量分析的定量數值差異有統計學意義(x2=187.900,P＜0.01),其中熒光定量PCR的定量數值最高,EMA-熒光定量PCR的數值次之,平闆培養的數值最低.結論 熒光定量PCR和EMA-熒光定量PCR方法檢測溫泉水中軍糰菌的靈敏度優于傳統的培養方法,EMA-熒光定量PCR方法適閤用于環境水體中軍糰菌活菌鑑測.
목적 비교전통적평판배양법、형광정량PCR화형광염료첩담추화을정(ethidium monoazide,EMA)결합형광정량PCR적방법(EMA-형광정량PCR)대온천수중군단균적검측효과.방법 우2011년매월(제5월빈)대북경시모온천도가촌적5개온천지진행채양,매월채집11빈수양,전년공채집121빈.분별채용전통적평판배양법、형광정량PCR화EMA-형광정량PCR방법대수양중군단균진행정성화정량분석.결과 평판배양법、형광정량PCR화EMA-형광정량PCR검측온천수중군단균오염솔분별위74.4％ (90/121)、100.0％(121/121)화100.0％(121/121)；3충방법대121빈온천수양본중군단균정량수치분별위0.10 ～ 216.00균락형성단위(CFU)/ml、1.47～1557.75기인단위(GU)/ml화0.20～301.69 GU/ml.형광정량PCR、EMA-형광정량PCR화평판배양방법대121빈온천수매빈군단균함량검측적중위수(제25백분위수～제75백분위수)분별위75.30 (32.51 ～ 192.10) GU/ml、36.46(16.08 ～91.21) GU/ml화5.30 (0.00 ～ 33.70) CFU/ml.대우121빈온천수수양,3충방법대군단균함량분석적정량수치차이유통계학의의(x2=187.900,P＜0.01),기중형광정량PCR적정량수치최고,EMA-형광정량PCR적수치차지,평판배양적수치최저.결론 형광정량PCR화EMA-형광정량PCR방법검측온천수중군단균적령민도우우전통적배양방법,EMA-형광정량PCR방법괄합용우배경수체중군단균활균감측.
Objective To compare the detection effect of Legionella 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 (x2 =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 plating method.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.
