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肺炎衣原体快速检测的环介导等温扩增方法的建立与评价

Development and evaluation of a loop-mediated isothermal amplification method for the rapid detection of Chlamydophila pneumoniae.

作者信息

Kawai Y, Miyashita N, Kishi F, Tabuchi M, Oda K, Yamaguchi T, Kawasaki K, Yamazaki T, Ouchi K

机构信息

Department of Pediatrics, Kawasaki Medical School, Okayama, Japan.

出版信息

Eur J Clin Microbiol Infect Dis. 2009 Jul;28(7):801-5. doi: 10.1007/s10096-009-0710-z. Epub 2009 Feb 11.

Abstract

We developed a loop-mediated isothermal amplification (LAMP) method to detect Chlamydophila pneumoniae infection. This assay exclusively amplified C. pneumoniae sequences and no cross-reactivity was observed for other Chlamydia species. The detection limit for this assay was found to be ten elementary bodies in 25 min, as observed in a real-time turbidimeter and electrophoretic analysis. The specificity of the LAMP reaction was confirmed by restriction endonuclease analysis, as well as direct sequencing of the amplified product. Among nasopharyngeal swab specimens from 120 patients with acute respiratory tract infections and 40 healthy individuals, the LAMP results showed 100% agreement with the results of real-time polymerase chain reaction (PCR) assays.

摘要

我们开发了一种环介导等温扩增(LAMP)方法来检测肺炎衣原体感染。该检测方法仅扩增肺炎衣原体序列,未观察到与其他衣原体物种的交叉反应。通过实时浊度仪和电泳分析发现,该检测方法的检测限为25分钟内10个原体。LAMP反应的特异性通过限制性内切酶分析以及扩增产物的直接测序得到证实。在120例急性呼吸道感染患者和40名健康个体的鼻咽拭子标本中,LAMP结果与实时聚合酶链反应(PCR)检测结果100%一致。

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