Mosquito Blood Meals: Identification by a Fluorescent Antibody Method

Roger M. McKinney Technical Development Laboratories, Malaria Program, Center for Disease Control, Arboviral Disease Section, Ecological Investigations Program, Center for Disease Control, Savannah, Georgia 31402

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Janet T. Spillane Technical Development Laboratories, Malaria Program, Center for Disease Control, Arboviral Disease Section, Ecological Investigations Program, Center for Disease Control, Savannah, Georgia 31402

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Preston Holden Technical Development Laboratories, Malaria Program, Center for Disease Control, Arboviral Disease Section, Ecological Investigations Program, Center for Disease Control, Savannah, Georgia 31402

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A procedure was developed for specific fluorescent antibody staining of erythrocytes from various mammalian species for the purpose of identifying the source of mosquito blood meals. Antibodies were prepared in rabbits against the erythrocyte membranes of humans, cows, horses, pigs, and sheep. The serum globulin fractions were labeled either with fluorescein isothiocyanate or tetramethylrhodamine isothiocyanate, each of which produced brilliant fluorescent antibody staining of homologous erythrocytes. Although initially cross-reactive, the conjugates were rendered specifically reactive, among the species tested, by absorption with stroma preparations of erythrocytes from the heterologous species. A blood meal extracted from a single mosquito, when diluted with saline, was sufficient to make eight microscope slide preparations. By pairing and mixing fluorescein labeled antibody to erythrocytes of one species with rhodamine labeled antibody to erythrocytes of another species, it was possible to test for two species with a single microscope slide preparation, thus allowing one to screen for 16 animal species with a blood meal extracted from a single mosquito. Blood meals were identified by this technique up to 44 hours after ingestion by the mosquito.

Author Notes

Present address: Laboratory Division, Center for Disease Control, Atlanta, Georgia 30333.

Dr. Holden died 17 May 1972.

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