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Am. J. Trop. Med. Hyg., 80(6), 2009, pp. 998-1003
Copyright © 2009 by The American Society of Tropical Medicine and Hygiene

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Identification of Flea Blood Meals Using Multiplexed Real-Time Polymerase Chain Reaction Targeting Mitochondrial Gene Fragments

Michael E. Woods*, John A. Montenieri, Rebecca J. Eisen, Nordin S. Zeidner, Jeff N. Borchert, Anne Laudisoit, Nackson Babi, Linda A. Atiku, Russell E. Enscore, AND Kenneth L. Gage
Bacterial Diseases Branch, Division of Vector-Borne Infectious Diseases, National Center for Zoonotic, Vector-Borne, and Enteric Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado; University of Antwerp, Antwerp, Belgium; Uganda Virus Research Institute, Entebbe, Uganda

Human plague is found in the West Nile region of Uganda and Democratic Republic of the Congo where flea vectors are often found inhabiting homes. We have developed a multiplexed, real-time polymerase chain reaction assay targeting mitochondrial genes that is capable of detecting blood meal sources in fleas collected off-host in East Africa. Laboratory tests showed that the assay is specific for the intended targets and has a detection limit below one picogram of DNA. Testing of wild-caught fleas from the Democratic Republic of Congo suggests that humans are at significant risk from flea-borne disease and implicates domestic animals including cats, chickens, and the black rat as potential sources of human exposure to fleas and flea-borne diseases. Future application of the assay will help us better define the ecology of plague in East Africa to implement effective control measures to combat the spread of disease.

Received November 25, 2008. Accepted for publication March 25, 2009.

Acknowledgments: Financial support: We acknowledge support from the Association of Public Health Laboratories Emerging Infectious Diseases Post-Doctoral Research Fellowship Program (M.E.W.) and Funds for Research in Industry and Agriculutre, Belgium (A.L.).

* Address correspondence to Michael E. Woods, Bacterial Diseases Branch, Division of Vector-Borne Infectious Diseases, NCZVED/ CDC, 3150 Rampart Rd., Fort Collins, CO 80521. E-mail: gvi1{at}cdc.gov

Authors’ addresses: Michael E. Woods, John A. Montenieri, Rebecca J. Eisen, Nordin S. Zeidner, Jeff N. Borchert, Russell E. Enscore, and Kenneth L. Gage, Bacterial Diseases Branch, Division of Vector-Borne Infectious Diseases, NCZVED/CDC, 3150 Rampart Rd., Fort Collins, CO 80521, Tel: 970-266-3584, Fax: 970-225-4257, E-mail: gvi1{at}cdc.gov. Anne Laudisoit, CERVA-CODA, 99, Groeselenberg, B-1180 Brussels, University of Antwerp, Evolutionary Ecology group, 171, Groenenborgerlaan, B-2020 Antwerpen and University of Liège, Zoogeography research unit, 27, Bd du Rectorat, B-4000 Liège, Belgium, Tel: ++3223790454, E-mail: anne.laudisoit{at}var.fgov.be. Nackson Babi and Linda A. Atiku, Uganda Virus Research Institute, PO Box 49, Entebbe Uganda, Tel: 256-041320385, E-mail: plague{at}ug.cdc.gov.






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Copyright © 2009 by the American Society of Tropical Medicine and Hygiene.