Detection of Yersinia pestis using real-time PCR in patients with suspected bubonic plague
Detection of Yersinia pestis using real-time PCR in patients with suspected bubonic plague
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DOI：
10.1016/j.mcp.2010.09.002
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发表时间：
2011-02-01
影响因子：
3.3
通讯作者：
Tomaso, Herbert
中科院分区：
文献类型：
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作者：
Riehm, Julia M.;Rahalison, Lila;Tomaso, Herbert
鼠疫耶尔森氏菌 (Y.) 是鼠疫的病原体，在亚洲、非洲和美洲的自然疫源地流行。实时 PCR 检测被描述为快速诊断工具，但迄今为止尚未对其临床应用进行验证。在一项回顾性临床研究中，我们评估了两种不同检测形式（5'-核酸酶检测和杂交探针检测）的三种实时 PCR 检测。研究人员对 149 名马达加斯加临床诊断为腺鼠疫患者的淋巴结抽吸物进行了鼠疫耶尔森氏菌 DNA 检测。将针对毒力质粒 pPCP1（pia 基因）和 pMT1（call，Ymt 基因）的实时 PCR 测定结果与 F1 抗原免疫层析测试 (ICT) 和生物体培养进行比较。在 149 个样本中，47 名患者经培养证实感染鼠疫杆菌，而 88 名患者 ICT 呈阳性，包括所有培养证实的病例。最好的实时 PCR 检测是针对 pla 的 5'-核酸酶检测，在 120 例病例中呈阳性。总之，当临床怀疑鼠疫时，可以推荐针对 pia 的 5'-核酸酶测定作为诊断工具，用于建立推定诊断。 (C) 2010 Elsevier Ltd. 保留所有权利。
Yersinia (Y.) pestis, the causative agent of plague, is endemic in natural foci of Asia, Africa, and America. Real-time PCR assays have been described as rapid diagnostic tools, but so far none has been validated for its clinical use. In a retrospective clinical study we evaluated three real-time PCR assays in two different assay formats, 5'-nuclease and hybridization probes assays. Lymph node aspirates from 149 patients from Madagascar with the clinical diagnosis of bubonic plague were investigated for the detection of Y. pestis DNA. Results of real-time PCR assays targeting the virulence plasmids pPCP1 (pia gene), and pMT1 (call, Ymt genes) were compared with an F1-antigen immunochromatographic test (ICT) and cultivation of the organism. Out of the 149 samples an infection with Y. pestis was confirmed by culture in 47 patients while ICT was positive in 88 including all culture proven cases. The best real-time PCR assay was the 5'-nuclease assay targeting pla which was positive in 120 cases. In conclusion, the 5'-nuclease assay targeting pia can be recommended as diagnostic tool for establishing a presumptive diagnosis when bubonic plague is clinically suspected. (C) 2010 Elsevier Ltd. All rights reserved.