16S rRNA PCR基因测序鉴定临床标本军团菌

Detection of legionella species in clinical specimens using PCR with sequencing confirmation

目的建立军团菌PCR基因测序鉴定方法,探讨广东地区非典型性肺炎患者军团菌感染状况。方法 2008年7月至2009年6月对广东地区各社区医院采集的100例非典型性肺炎痰液标本,采用军团菌16S rRNA基因片段作PCR试验,基因测序法对PCR结果进行鉴定。结果 100例非典型性肺炎患者痰液16S rRNA PCR试验阳性13例,其中12例经测序鉴定为嗜肺军团菌,1例测序结果阴性,结果表明PCR试验敏感性100%,特异性98.86%。结论 16S rRNA PCR基因测序不仪可以提高阳性率,而且可以排除假阳性。本研究结果显示,广东地区各社区医院内非典型性肺炎患者的军团菌感染率为1 2%。

Objective To establish the methods of PCR-gene sequencing of Legionella species and to investigate patients with pneumonia of legionella infection in Guangdong region. Methods 100 sputum samples of patients with atypical pneumonia in Guangdong region were investigated by PCR-sequencing of the 16S rRNA fragment of legionella. Results 13 cases were positive of polymerase chain reaction of 16S rRNA fragment,among 12 cases were identified as legionella pneumophila by sequencing. The sequencing result of one cases has no signal. The results shown that the sensitivity and specificity of PCR was 100% and 98.86% ,respectively. Conclusion The polymerase chain reaction of 16S rRNA is a recognized method of molecular detection of legionella,It can been not only improved the positive rate,but also ruled out the false positive. The results shown that infection rate of the patient with atypical pneumonia of community hospital in Guangdong region was 12%.