Immunoenzymatic Labeling of Multiple Plasmodial Salivary Gland Sporozoites in a Single Test

Claudia F. GolendaDepartments of Entomology and Immunology, Division of Communicable Diseases and Immunology, Walter Reed Army Institute of Research, Washington, DC

Search for other papers by Claudia F. Golenda in
Current site
Google Scholar
PubMed
Close
,
Ted HallDepartments of Entomology and Immunology, Division of Communicable Diseases and Immunology, Walter Reed Army Institute of Research, Washington, DC

Search for other papers by Ted Hall in
Current site
Google Scholar
PubMed
Close
,
Imogene SchneiderDepartments of Entomology and Immunology, Division of Communicable Diseases and Immunology, Walter Reed Army Institute of Research, Washington, DC

Search for other papers by Imogene Schneider in
Current site
Google Scholar
PubMed
Close
, and
Robert A. WirtzDepartments of Entomology and Immunology, Division of Communicable Diseases and Immunology, Walter Reed Army Institute of Research, Washington, DC

Search for other papers by Robert A. Wirtz in
Current site
Google Scholar
PubMed
Close
Restricted access

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.

Save