title: Immunoenzymatic labeling of multiple plasmodial salivary gland sporozoites in a single test.
abstract: A direct, double- and triple-staining immunoenzymatic method detected and differentiated sporozoites by color in Anopheles stephensi 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 Plasmodium berghei and P. falciparum 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 P. falciparum and two strains of P. vivax sporozoites. Monoclonal antibodies to the CS proteins conjugated to beta-galactosidase (P. falciparum), alkaline phosphatase (P. vivax variant), and horseradish peroxidase (P. vivax 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.
authors.affiliation: Department of Entomology, Walter Reed Army Institute of Research, Washington, DC.
authors.forename: C F
authors.initials: CF
authors.lastname: Golenda
authors.forename: T
authors.initials: T
authors.lastname: Hall
authors.forename: I
authors.initials: I
authors.lastname: Schneider
authors.forename: R A
authors.initials: RA
authors.lastname: Wirtz
mesh_terms: D000818:Animals; D000852:Anopheles / Q000469:parasitology*; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D007303:Insect Vectors / Q000469:parasitology*; D010962:Plasmodium berghei / Q000302:isolation & purification*; D010963:Plasmodium falciparum / Q000302:isolation & purification*; D010966:Plasmodium vivax / Q000302:isolation & purification*; D012469:Salivary Glands / Q000469:parasitology