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 clin ical samples. We developed a 5' nuclease PCR assay targeting the plasm inogen activator gene (pla) of Yersinia pestis. The assay is species s pecific, with a detection threshold of 2.1 x 10(5) copies of the pla t arget or 1.6 pg of total cell DNA. The assay detected Y. pestis in exp erimentally infected Xenopsylla cheopis fleas and in experimentally in fected monkey blood and oropharyngeal swabs. The TaqMan assay is simpl e to perform and rapid and shows promise as a future field-adaptable t echnique.