Immunodot assay of Plasmodium falciparum sporozoites in mosquitoes using a direct binding membrane system.
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.
Long
G W
GW
Infectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland.
Charoenvit
Y
Y
Oprandy
J J
JJ
D000818:Animals; D000852:Anopheles; D000913:Antibodies, Protozoan / Q000276:immunology; D000953:Antigens, Protozoan / Q000276:immunology / Q000302:isolation & purification*; D015151:Immunoblotting / Q000379:methods; D008567:Membranes, Artificial; D010270:Parasite Egg Count; D010963:Plasmodium falciparum / Q000032:analysis* / Q000276:immunology; D011485:Protein Binding; D015800:Protozoan Proteins / Q000276:immunology / Q000302:isolation & purification*
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D008567:Membranes, Artificial; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan