Quantitative determination of sporozoites and circumsporozoite antigen in mosquitoes infected with Plasmodium falciparum or P-vivax

It is essential for malariologists and researchers to have simple and accurate means of assessing the threat of Plasmodium parasites. An attempt was therefore made to re-standardize one of the circumsporozoite (CS) ELISA that can be used to detect and quantify the circumsporozoite antigens of P. falciparum and P. vivax. A two-site, 'sandwich' ELISA based on a monoclonal antibody was used to test for the CS antigen and sporozoites of each Plasmodium species simultaneously. Using the resultant optical-density values, standard curves, that permit the number of sporozoites in an infected mosquito to be estimated from the quantification of the CS antigen, were constructed. Using these plots and the CS ELISA, the presence of just 12.5 sporozoites (i.e. 0.8 pg CS antigen) of P. falciparum, four sporozoites (3.2 pg antigen) of P. vivax-210 or 12.5 sporozoites (32.0 pg antigen) of P. vivax-247 could be demonstrated.