鼠疫耶尔森菌的多重PCR检测方法

Detection of Yersinia pestis by multiplex polymerase chain reaction

目的研究一种快速、特异的检测鼠疫耶尔森菌(鼠疫菌)的多重聚合酶链反应(PCR)方法。方法选用针对鼠疫菌F1抗原基因、pla基因、inv基因和一段鼠疫特异染色体序列设计的4对引物,以鼠疫菌及其他肠道致病菌DNA为模板并加入内部对照模板,在同一扩增体系中进行PCR扩增。结果鼠疫菌DNA模板可扩增出预期产物,而其他菌株只能扩增出内部对照模板的片段。结论多重PCR方法具有较高的特异性。可迅速、有效地将鼠疫菌与其他肠道致病菌相鉴别,也可应用于鼠疫监测。

Objective To apply a rapid and special method for the detection of Yersinia pestis by multiplex polymerase chain reaction（PCR）. Methods The F1 antigen, pla gene, inv gene and a specific segment on the chromosome were co-amplified by PCR. An internal control（IC） was required in order to prevent false negative results that might be caused by PCR inhibitors. Other enteric pathogenic bacteria DNA were amplified as control. Results The amplicons of Yersinia pestis DNA were the same with the anticipative products. Only internal control amplicon was gotten while amplifying other enteric pathogenic bacteria DNA. Conclusion This method is specific and rapid and can identify Yersinia pestis. It is useful for detection of Yersinia pestis.