双引物DNA扩增检测成人社区获得性肺炎中的支原体感染

Detection of Mycoplasma pneumoniae in Community Acquired Pneumonia by Two Primer Polymerase Chain Reaction

目的 研究肺炎支原体在社区获得性肺炎 (CAP)中的重要性及快速检测肺炎支原体的PCR方法。方法 收集 2 0 0 1年 11月～ 2 0 0 2年 6月社区获得性肺炎患者共 113例 ;取患者急性期痰标本 (无痰者留取咽拭子 )进行检测 ;先扩增针对肺炎支原体特异性较强的P1粘附蛋白基因 ,同时应用UDG消除污染 ,减少假阳性 ,并加用内参照避免假阴性 ;P1基因扩增阳性者再行有种属特异性的 16SrRNA基因扩增进行验证。结果 113例CAP患者中共有 2 9例 (2 5 7% )P1基因及 16SrRNA基因检测皆为阳性。结论 采用准确性较高的双引物PCR扩增方法 ,证明在我国肺炎支原体是成人CAP的主要致病原。

OBJECTIVE To investigate the importance of Mycoplasma pneumoniae in community acquired pneumonia(CAP) and the rapid diagnosis of M. pneumoniae infection by polymerase chain reaction (PCR). METHODS A prospective study was performed on 113 consecutive adult patients with CAP from Nov 2001 to June 2002. Sputum samples (or throat specimens) in acute period were collected for the test.The primers directed against the P1 adhesion gene were first used in the PCR. In the reaction, the uracil-DNA-glycosylase (UDG) system was used to prevent contamination from previous amplifications, and a specific internal control was added to detect possible inhibitors of the reaction. When the results were positive, the samples were confirmed with the PCR using the primers directed against species-specific sequence of the 16S ribosome gene. RESULTS PCR was positive for both P1 and the 16S gene in 29 samples of patients with CAP. CONCLUSIONS By two primer PCR method,we confirm that M. pneumoniae is an important pathogen in adult patients with CAP.