A rapid sporozoite ELISA using 3,3',5,5'-tetramethylbenzidine as the substrate chromogen.
A modified version of the standard 2-site sporozoite enzyme-linked immunosorbent assay (ELISA) using 3,3',5,5'-tetramethylbenzidine (TMB) as the substrate chromogen solution was adapted for rapid detection and identification of Plasmodium falciparum and P. vivax circumsporozoite (CS) proteins. The TMB-ELISA was evaluated using sporozoites from experimentally infected mosquitoes and laboratory colonized uninfected mosquitoes. Our data indicate comparable sensitivity levels between the TMB-ELISA and the standard ELISA, i.e., 50 P. falciparum or P. vivax sporozoites/50 microliters of test solution. Reactions inherent to the method were specific and background reactivity was minimal. The TMB-ELISA is rapid (1 hr), simple, uses a minimal amount of monoclonal antibodies, and is suitable for use in a wide range of laboratories.
U.S. Army Medical Research Unit, Institute for Medical Research, Kuala Lumpur, Malaysia.
M
M
Lee
B A
BA
Harrison
G E
GE
Lewis
D000818:Animals; D000852:Anopheles / Q000469:parasitology; D000953:Antigens, Protozoan / Q000032:analysis*; D001560:Benzidines; D002863:Chromogenic Compounds; D004797:Enzyme-Linked Immunosorbent Assay; D010963:Plasmodium falciparum / Q000302:isolation & purification*; D010966:Plasmodium vivax / Q000302:isolation & purification*; D011237:Predictive Value of Tests; D015800:Protozoan Proteins / Q000032:analysis*
D000953:Antigens, Protozoan; D001560:Benzidines; D002863:Chromogenic Compounds; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan; C021758:3,3',5,5'-tetramethylbenzidine