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IDENTIFICATION OF MAMMALIAN BLOOD MEALS IN MOSQUITOES BY A MULTIPLEXED POLYMERASE CHAIN REACTION TARGETING CYTOCHROME B

To date, no polymerase chain reaction diagnostic technique exists to directly identify mammalian blood meals from mosquitoes by sized DNA fragments following agarose gel electrophoresis. We have developed a vertebrate-specific multiplexed primer set based on mitochondrial cytochrome b to identify the mammalian blood hosts of field-collected mosquitoes. Although designed for the study of African malaria vectors, the application of this tool is not restricted to this disease system. Validation of this diagnostic technique on dried anopheline and culicine field specimens collected in Zambia and Mali demonstrated that blood meals could be identified 2–7 months after collection. Time course experiments showed that host DNA was detectable in frozen mosquito abdomens 24–30 hours post-feeding. Additionally, multiple blood meals from different mammals could be detected in a single mosquito. This diagnostic assay will be a valuable tool for identifying the blood meals of field-collected mosquitoes where people and alternative mammal hosts are present.

Received November 23, 2004. Accepted for publication March 31, 2005.

Acknowledgments: We thank Dr. Sungano Mharakurwa and Dr. Phil Thuma for their time and effort spent coordinating mosquito collections and field team operations in Zambia, as well as Harry Hamapumbu, Patricia Muleya, Petros Moono, Fidelis Chanda, Lushomo Chikobolo, Collence Munsanje, Rodwell Moono, Peter Simakwati, Guide Hansumo, Seen Mudenda, Betham Dubeka, and the Macha Malaria Research Institute staff for their assistance. We also thank Dr. Guimogo Dolo for his assistance with collections in Mali.

Financial support: This research was supported in part by financial assistance to Douglas E. Norris from the Johns Hopkins Malaria Research Institute, an award from the United National Development Program/World Bank/World Health Organization Special Program for Research and Training in Tropical Diseases (TDR) (A10360) to Douglas E. Norris, a Johns Hopkins Malaria Research Institute pre-doctoral fellowship award to Rebekah J. Kent, and a National Institute of Environmental Health Sciences training award (T32ES07141) to Rebekah J. Kent.

^* Address correspondence to Dr. Douglas E. Norris, W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205. E-mail: dnorris{at}jhsph.edu

Authors’ address: Rebekah J. Kent and Douglas E. Norris, W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205, Telephone: 410-614-2710 Fax: 410- 955-0105, E-mails: rkent{at}jhsph.edu and dnorris{at}jhsph.edu.

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