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Am. J. Trop. Med. Hyg., 79(1), 2008, pp. 99-101
Copyright © 2008 by The American Society of Tropical Medicine and Hygiene

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SHORT REPORT


Development of a Real-time Quantitative PCR Assay to Enumerate Yersinia pestis in Fleas

Elizabeth S. Gabitzsch, Rommelle Vera-Tudela, Rebecca J. Eisen, Scott W. Bearden, Kenneth L. Gage, AND Nordin S. Zeidner*
Centers for Disease Control and Prevention, Division of Vector-Borne Infectious Diseases, Fort Collins, Colorado

 

ABSTRACT

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.

Received January 9, 2008. Accepted for publication March 25, 2008.

* Address correspondence to Nordin S. Zeidner, Centers for Disease Control and Prevention, Division of Vector-Borne Infectious Diseases, 3150 Rampart Road, Fort Collins, CO 80521. E-mail: Naz2{at}cdc.gov

Authors’ addresses: Elizabeth S. Gabitzsch, Rommelle Vera-Tudela, Rebecca J. Eisen, Scott W. Bearden, Kenneth L. Gage, and Nordin S. Zeidner, Centers for Disease Control and Prevention, Division of Vector-Borne Infectious Diseases, Fort Collins, CO, Tel: +1 (970) 221-6495, Fax: +1 (970) 225-4257, E-mail: Naz2{at}cdc.gov.






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