Volume 46, Issue 3
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



A direct, double- and triple-staining immunoenzymatic method detected and differentiated sporozoites by color in salivary glands and in mixed sporozoite slide preparations. A double-staining method used beta-galactosidase- and alkaline phosphatase-labeled monoclonal antibodies to the circumsporozoite (CS) proteins of and in mosquito salivary glands. The CS proteins were distinguished clearly by the blue-green and red substrate products of beta-galactosidase and alkaline phosphatase, respectively. A triple-staining method differentiated by color among a mixture of and two strains of sporozoites. Monoclonal antibodies to the CS proteins conjugated to beta-galactosidase (), alkaline phosphatase ( variant), and horseradish peroxidase ( predominant) readily color differentiated sporozoites by the blue-green, purple-blue, and orange-brown substrate products, respectively. This assay may have potential use in malaria transmission studies, genetic crosses of variant strains of plasmodia to determine assortment of CS antigen alleles, and as a technique to determine the fate of the CS antigen in infected mosquitoes.


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