基因扩增法检测军团杆菌DNA的研究

Detection of legionellae with polymerase chain reaction

采用聚合酶链式反应建立检测军团菌 DNA 的实验诊断方法。结果显示,采用军团菌属5 SrRNA 编码区特异性引物进行扩增,11种军团菌(包括八种嗜肺军团菌)均可见104bp 的阳性扩增带;采用嗜肺军团菌巨噬细胞感染增强(mip)基因编码区特异性引物扩增,所有八种嗜肺军团菌均可见650 bp 的特异性扩增带,所有三种非嗜肺军团菌扩增结果为阴性。检测敏感性为60 fg 左右。

We have developed the methods for detection of cutured legionella,based upon the polymerase chain reaction(PCR).These results showed that all 11 strains of legionellae(including all 8 serogroups of L.pneumophila)were detected by PCR amplification of a 104bp DNA sequence that codes for a region of 5SrRNA with genus-specific primers;and all 8 serogroups of L.pneumophila were detected by ampilification of a 650bp of the coding region DNA sequences of the macrophage infectivity potentiator(mip)gene by species-specific primers.Bands of positive amplified DNA were detected in 60fg template DNA.Our study suggested that this technique could be applied to clinical specimens for rapid and specific diagnosis of legionnairis disease,environmental monitoring and control of fulminant epidemic of legionellosis.