聚合酶链反应检测嗜肺军团菌的实验研究

Detection of Legionella pneumophila by polymerase chain reaction

目的探讨并建立聚合酶链反应(PCR)用于检测嗜肺军团菌的方法。方法用PCR方法扩增嗜肺军团菌巨噬细胞感染增强子基因(mip)片段,琼脂糖凝胶电泳检测扩增产物;分析其特异性和敏感性。结果在嗜肺军团菌14个血清型参考株均扩增出特异的340 bp片段,而三株非嗜肺军团菌及其他需鉴别诊断的常见病原菌均未扩增出此特异条带。检测灵敏度为2.6 pg/ul基因组DNA。结论依据mip基因建立的嗜肺军团菌PCR检测方法具有高度的敏感性和特异性,是诊断与鉴别诊断嗜肺军团菌感染的一种有效手段。

Objective To develop a rapid method for detecting Legionella pneumophila （Lp） by polymerase chain reaction（PCR）. Methods The specific fragment of maerophage infectivity potentiator （mip） gene of Lp was detected by specific polymerase chain reaction. PCR amplification products were examined by agarose gel electrophoresis. In addition, the specificity and sensitivity of this method were evaluated.Results The 340 bp specific fragment of mip gene was amplified in all the 14 tested Lp strains,whereas it was detected in none of the differential control strains including three non - pneamophila strains. The lowest limit of detection was 2.6 pg/μl genome DNA of Lp strain. Conclusion The PCR assay based on mip gene has high specificity and sensitivity and is valuable for the diagnosis and differential diagnosis of Lp when cultures are not available.