5′ Nuclease PCR Assay To Detect Yersinia pestis
James A. Higginsa b, John W. Ezzella b, B. Joseph Hinnebuschc d, M Shipleya b, Erik A. Henchala b, M. Sofi Ibrahima b
- a<!--label omitted: 1-->Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland 21702-5011,1 and
- bDiagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland 21702-5011,1 and
- c<!--label omitted: 2-->Laboratory of Microbial Structure and Function, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 598402
- dLaboratory of Microbial Structure and Function, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 598402
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Abstract
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 x 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.
TL;DRA 5′ nuclease PCR assay targeting the plasminogen activator gene (pla) ofYersinia pestis is developed, with a detection threshold of 2.1 × 105 copies of thepla target or 1.6 pg of total cell DNA. (Semantic Scholar)