Processing and microfiltration of mosquitoes for malaria antigen detection in a rapid dot immunobinding assay.
Data on a technique for the detection of antigen from arthropod vectors in a dot immunobinding assay are presented. In this system, antigen present in the vector was first solubilized in sodium dodecyl sulfate. The homogenate from this process was microfiltered through a two-membrane sandwich; target antigen molecules passed through the first membrane and were immobilized on the second one. The first membrane was nonbinding and served to impinge debris. The second membrane was a high-protein-binding-capacity hydrophobic polyvinylidene difluoride membrane. High signal-to-noise ratios were produced by this method, which is readily adaptable for field use. This assay was used for malaria sporozoites, but it can serve as a general technique that is applicable to other arthropod vectors and etiologic agents.
Oprandy
J J
JJ
Infectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland 20814.
Long
G W
GW
D000818:Animals; D000852:Anopheles / Q000469:parasitology; D000911:Antibodies, Monoclonal / Q000276:immunology; D000953:Antigens, Protozoan / Q000032:analysis*; D005374:Filtration; D015151:Immunoblotting; D008567:Membranes, Artificial; D010963:Plasmodium falciparum / Q000276:immunology*; D011145:Polyvinyls; D012680:Sensitivity and Specificity; D012995:Solubility; D005465:Fluorocarbon Polymers
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D008567:Membranes, Artificial; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers