5′ Nuclease PCR Assay To Detect Yersinia pestis

The 5′ nuclease PCR assay uses a fluorescently labeled oligonucleotide probe (TaqMan) to rapidly detect and quantitate DNA templates in clinical samples. We developed a 5′ nuclease PCR assay targeting the plasminogen activator gene (pla) of Yersinia pestis. The assay is species specific, with a detection threshold of 2.1 × 10(5) copies of the pla target or 1.6 pg of total cell DNA. The assay detected Y. pestis in experimentally infected Xenopsylla cheopis fleas and in experimentally infected monkey blood and oropharyngeal swabs. The TaqMan assay is simple to perform and rapid and shows promise as a future field-adaptable technique.

5'核酸酶PCR检测法利用荧光标记寡核苷酸探针（TaqMan），可快速检测并定量临床样本中的DNA模板。我们针对鼠疫耶尔森菌（Yersinia pestis）的纤溶酶原激活物基因（pla）开发了一种5'核酸酶PCR检测法。该检测法具有物种特异性，检测阈值为2.1×10^5个pla靶标拷贝，或1.6皮克（pg）的总细胞DNA。该检测法可在实验感染的印鼠客蚤（Xenopsylla cheopis）、实验感染的猴血液以及口咽拭子中检出鼠疫耶尔森菌。该TaqMan检测法操作简便、快速高效，有望成为未来可适配现场检测的技术手段。