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Am. J. Trop. Med. Hyg., 43(6), 1990, pp. 597-601
Copyright © 1990 by The American Society of Tropical Medicine and Hygiene

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Immunodot Assay of Plasmodium falciparum Sporozoites in Mosquitoes Using a Direct Binding Membrane System

Gary W. Long, Yupin Charoenvit AND John J. Oprandy
Infectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland

We describe a membrane based immunodot assay for the detection of Plasmodium falciparum sporozoites mixed with mosquitoes. A crude sodium dodecyl sulfate extract of mosquitoes and sporozoites is passed through a bi-layered membrane system, the top layer being a polyvinyldiene difluoride hydrophilic pre-filter which screens out debris but allows the passage of antigen. Sporozoite, as well as mosquito, proteins are bound to the hydrophobic membrane below. This membrane was probed with a monoclonal antibody to the repeat region of the P. falciparum circumsporozoite protein, a peroxidase labeled second antibody and a tetramethyl-benzidine substrate. The method detects as few as 10 sporozoites/mosquito or 100 sporozoites in a pool of 10.






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Copyright © 1990 by the American Society of Tropical Medicine and Hygiene.