Bundeswehr Institute of Microbiology, Munich, Germany.
Mol Cell Probes. 2011 Feb;25(1):8-12. doi: 10.1016/j.mcp.2010.09.002. Epub 2010 Oct 8.
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 (pla gene), and pMT1 (caf1, 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 pla can be recommended as diagnostic tool for establishing a presumptive diagnosis when bubonic plague is clinically suspected.
鼠疫耶尔森菌（Y. pestis）是鼠疫的病原体，在亚洲、非洲和美洲的自然疫源地中流行。实时 PCR 检测已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'-核酸酶和杂交探针检测）的三种实时 PCR 检测方法。对马达加斯加 149 例临床诊断为腺鼠疫的患者的淋巴结抽吸物进行了鼠疫耶尔森菌 DNA 的检测。针对毒力质粒 pPCP1（pla 基因）和 pMT1（caf1、Ymt 基因）的实时 PCR 检测结果与 F1-抗原免疫层析检测（ICT）和病原体培养进行了比较。在 149 个样本中，47 例通过培养证实了鼠疫耶尔森菌感染，而 ICT 在 88 例中呈阳性，包括所有培养阳性病例。最有效的实时 PCR 检测方法是针对 pla 的 5'-核酸酶检测，该检测在 120 例中呈阳性。总之，当临床怀疑腺鼠疫时，针对 pla 的 5'-核酸酶检测可作为建立推定诊断的诊断工具推荐使用。
