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Am. J. Trop. Med. Hyg., 71(4), 2004, pp. 514-517
Copyright © 2004 by The American Society of Tropical Medicine and Hygiene

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MOLECULAR DIFFERENTIATION OF COLONIZED HUMAN MALARIA VECTORS BY 28S RIBOSOMAL DNA POLYMORPHISMS

REBEKAH J. KENT, AIMEE J. WEST, AND DOUGLAS E. NORRIS
The W. Harry Feinstone Department of Molecular Microbiology and Immunology, The Johns Hopkins Malaria Research Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland

Anopheles gambiae s.s. Giles, An. stephensi Liston, An. freeborni Aitken, and An. quadrimaculatus Say are cultured and studied in molecular genetic and transgenic laboratories with increasing frequency. With limited research space, these mosquitoes are often maintained in the same insectary. Under these conditions, cross-contamination of colonies can occur and have devastating consequences to affected research programs. We have developed a polymerase chain reaction-based assay targeting the 28S large subunit ribosomal RNA gene to easily differentiate between these four taxa and An. funestus Giles, which occurs in sympatry with An. gambiae. The resulting assay identifies individual mosquito preparations as well as all taxa within a mixed or pooled DNA template preparation.

Received December 24, 2003. Accepted for publication April 14, 2004.

Acknowledgments: We thank M. Coetzee and R. Hunt for providing insight into the morphology of these anopheline taxa and providing An. funestus colony specimens. We also thank the laboratories of N. Kumar, M. Jacobs-Lorena, and S. Luckhart for providing colony mosquitoes for this study, and S. Shone for donating field-collected An. quadrimaculatus.

Financial support: This research was supported by financial assistance to Douglas E. Norris from the UNDP/World Bank/World Health Organization Special Program for Research and Training in Tropical Diseases (TDR) (A10360), partial support from the Johns Hopkins Malaria Research Institute, and a NIEHS training award (T32ES07141) to Rebekah J. Kent.

Authors’ address: Rebekah J. Kent, Aimee J. West, and Douglas E. Norris, The 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-mail: dnorris{at}jhsph.edu.






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Copyright © 2004 by the American Society of Tropical Medicine and Hygiene.