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为探讨双重PCR方法 (DPCR)检测痰标本中军团菌DNA在早期诊断军团菌肺炎方面的意义 ,采用DPCR方法对军团菌肺炎组、临床疑似军团菌肺炎组、普通肺炎组和肺结核组 4组患者留取的痰标本进行检测。DPCR过程采用根据军团菌 1 6SrRNA基因和mip基因设计的 2对引物 ,同时扩增军团菌 3 86bp 1 6SrRNA基因片段和 2 0 6bpmip基因片段。结果 :军团菌肺炎组的DPCR阳性检出率为 1 0 0 % ,明显高于其他 3组 (分别为 1 7.2 4%、0、0 ,P均 <0 .0 1 )。模拟痰标本DPCR的最低检出限为 1× 1 0 3 cfu/mL。初步研究表明 ,DPCR方法检测痰标本中的军团菌DNA具有较好的敏感性、特异性和稳定性 ,此法在临床早期诊断军团菌肺炎方面具有一定的价值
To investigate the significance of using the double PCR method (DPCR) to detect Legionella pneumophila DNA in sputum specimens for the early diagnosis of Legionella pneumonia, four groups of patients with Legionella pneumophila pneumonia, clinical suspected Legionnaires pneumonia, common pneumonia group and pulmonary tuberculosis group were treated with DPCR method Sputum specimens taken for testing. In the DPCR process, two pairs of primers designed according to Legionella 16SrRNA and mip genes were used to amplify the 3 86bp 16SrRNA gene fragment and the 20 6bpmip gene fragment of Legionella simultaneously. Results: The positive rate of DPCR in Legionella pneumonia group was 100%, which was significantly higher than the other three groups (1 7.24%, 0,0, P <0.01). The minimum detection limit of DPCR in simulated sputum specimens was 1 × 10 3 cfu / mL. Preliminary studies have shown that the DPCR method for detection of Legionella DNA in sputum specimens has a good sensitivity, specificity and stability, this method has some value in early clinical diagnosis of Legionella pneumonia