Gabitzsch ES, Vera-Tudela R, Eisen RJ, Bearden SW, Gage KL, Zeidner NS
Development of a real-time quantitative PCR assay to enumerate Yersinia pestis in fleas. [Journal Article]
Am J Trop Med Hyg 2008 Jul; 79(1):99-101.
A real-time quantitative polymerase chain reaction (qPCR) assay was developed for Yersina pestis. The qPCR assay was developed utilizing a conserved region of the Y. pestis ferric iron uptake regulator gene (fur) to design primers and a fluorescent (FAM-labeled) TaqMan probe. The assay was optimized using cultured Y. pestis (UG05-0454) and was confirmed to work with strains from 3 Y. pestis biovars. The optimized assay was capable of detecting a single organism of cultured Y. pestis and as little as 300 bacteria in infected flea triturates. This qPCR assay enables rapid enumeration of Y. pestis bacterium in laboratory-infected fleas when compared with conventional serial dilution plating.
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