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

Gary W. LongInfectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland

Search for other papers by Gary W. Long in
Current site
Google Scholar
PubMed
Close
,
Yupin CharoenvitInfectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland

Search for other papers by Yupin Charoenvit in
Current site
Google Scholar
PubMed
Close
, and
John J. OprandyInfectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland

Search for other papers by John J. Oprandy in
Current site
Google Scholar
PubMed
Close
Restricted access

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.

Save