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首页> 外文期刊>Acta microbiologica et immunologica Hungarica: A quarterly of the Hungarian Academy of Sciences >Single tube real time PCR for detection of Streptococcus pneumoniae, Mycoplasma pneumoniae, Chlamydophila pneumoniae and Legionella pneumophila from clinical samples of CAP
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Single tube real time PCR for detection of Streptococcus pneumoniae, Mycoplasma pneumoniae, Chlamydophila pneumoniae and Legionella pneumophila from clinical samples of CAP

机译：单管实时PCR检测链球菌肺炎链球菌，支原体肺炎，衣原体肺炎肺炎肺炎肺炎（肺炎

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摘要

We designed a multiplex real time PCR for rapid, sensitive and specific detection of Streptococcus pneumoniae, Legionella pneumophila, Chlamydophila pneumoniae and Mycoplasma pneumoniae. The study cases consisted of 129 patients with community acquired pneumonia (CAP). Bacteriological techniques were implemented for detection of the cultivable organisms. DNA were extracted from sputa, throat swabs, bronchoalveolar lavages and tracheal aspirates and used as templates in real time PCR. The primers and probes were designed for cbpA (S. pneumoniae), p1adhesin (M. pneumoniae), mip (L. pneumophila) and ompA (C. pneumoniae). After optimization of real time PCR for every organism, the experiments were continued in multiplex in a single tube. Of 129 CAP specimens, the positive cultures included 14 (10.85%) for S. pneumoniae, 9 (6.98%) for L. pneumophila and 3 (2.33%) for M. pneumoniae. Four specimens (3.10%) were positive for C. pneumoniae by real time PCR. The sensitivity of our real time PCR was 100% for all selected bacteria. The specificity of the test was 98.26%, 98.34%, 100% and 100% for S. pneumoniae, L. pneumophila, M. pneumoniae and C. pneumoniae, respectively. This is the first report on the use of multiplex real time PCR for detection of CAP patients in the Middle East. The method covers more than 90% of the bacterial pathogens causing CAP.
机译：我们设计了一种用于快速，敏感和特异性检测的多重实时PCR，肺炎链球菌，军团菌肺炎，衣原体肺炎和支原体肺炎。研究病例由129名患有129名患有肺炎（帽）的患者组成。实施了用于检测可耕种生物的细菌学技术。从丁塔，喉拭子，支气管肺泡灌洗和气管脱臼中提取DNA，并在实时PCR用作模板。设计引物和探针是针对CBPA（肺炎），P1Adhesin（M.Pneumoniae），MIP（L.Pneumophila）和OMPA（C.Pneumoniae）的CBPA。在每种生物的优化实时PCR后，在单管中继续进行实验。在129个帽子标本中，阳性培养物包括14（10.85％）的S.肺炎，9（6.98％）为L.Pneumophila和3（2.33％）的M.肺炎。通过实时PCR为C.肺炎的四个标本（3.10％）为阳性。所有选定的细菌的实时PCR的敏感性为100％。 Spneumoniae，L.Pneumophila，M.Pneumophila，M.肺炎和C.肺炎的特异性，该试验的特异性分别为98.26％，98.34％，100％和100％。这是关于使用多重实时PCR用于检测中东概念患者的第一个报告。该方法占据了超过90％的细菌病原体导致帽。

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