基因扩增法检测军团杆菌DNA的研究

Detection of Legionellae with Polymerase Chain Reaction

本研究采用聚合酶链式反应（PCR），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5SrRNA编码区特异性引物进行扩增，11种军团菌（包括8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8种嗜肺军团菌均可见650bp的特异性扩增带，所有3种非嗜肺军团菌扩增结果为阴性。检测敏感性为60fg左右。该研究技术的建立，为军团病的早期诊断和环境监测及控制爆发流行提供了一种快速、敏感、特异的新方法。

We have developed the method for detection of cultured legionella, based upon the polymerase chain reaction (PCR). These results showed that all 11 strains of legionellas (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 seuqences 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 applide to clinical specimens for rapid identification and specific diagnosis of legionella disease,so that environmental monitoring and control of fulminant epidendc of legionellosis could be possible.