应用双重PCR检测环境水体中的军团菌

建立双重PCR方法以检出环境水体中的军团菌。设计两对引物,分别扩增军团菌的16S rRNA和M ip基因,扩增片段长各为375bp和996bp。该方法检测军团菌的灵敏度为5.8×102cfu/m l,6株嗜肺标准军团菌均扩增出996bp和375bp两条带,4株非嗜肺军团菌扩增出375bp条带,4株非军团菌无条带;检测71份环境水样,5份出现两条条带,2份可见375bp条带,阳性率为7.0%。该方法快速、灵敏、特异,为水体中的嗜肺军团菌检测提供了有效方法。

Development of a duplex PCR assay for detecting Legionella in environmental waters

A duplex PCR assay is established to detect Legionella in environmental waters. Two different sets of oligouncleotide primer were used simultaneously to amplify 375bp from 16S rRNA gene and 996bp from Mip gene. The sensitivity of this assay is 5.8×10~2cfu/ml, 375bp and 996bp fragments can be amplifed from 6 reference strains of Legionella pneumophila and 375bp from 4 strains of non-Legionella pneumophila, no DNA product was observed in 4 non-Legionella strains. Of 71 water sampels, 996bp and 375bp fragments were observed in 5 samples, only 375bp fragment in 2 samples, the positive rate is 7.0%. This assay has high sensitivity and specificity and offers an effective method to detect Legionella pneumophila in waters.