聚合酶链法检测环境水体中嗜肺军团菌

Detection of legionella pneumophila with polymerase chain reaction in environmental waters

目的建立检测环境水体中嗜肺军团菌聚合酶链(PCR)法。方法以嗜肺军团菌特异性保守基因Mip为靶基因,采用Godkex软件设计引物,聚合酶链反应测定。结果该方法检测军团菌的灵敏度为5.8×102cfu/ml,6株标准军团菌均扩增出996 bp的条带,4株非嗜肺军团菌和4株非军团菌无条带出现;检测71份环境水样,5份阳性,阳性率为7.0%。结论该方法快速、灵敏、特异,是检测水体中嗜肺军团菌的有效方法。

Objective To develop an approach for the detection of legionella pneumophila with polymerase chain reaction(PCR).Methods Assay was established to detect legionella pneumophila in environmental waters.Results The sensitivity of this assay was 5.8×10~2 cfu/ml.A 996-bp DNA fragment was amplified from 6 reference legionella pneumophila strains,and no DNA product was observed in non-Legionella pneumophila strains and 4 non-Legionella strains.5 samples were observed positive in 71 samples,and the positive rate wais 7.0%. Conclusions This assay has high sensitivity and specificity and offers an effective method to detect legionella pneumophila in waters.