A nitrocellulose membrane based solid phase enzyme immunoassay for detecting polioviruses in stool.
An enzyme immunoassay was developed with nitrocellulose membrane as the solid phase support built in a 96 well porous polystyrene plate. Monoethanolamine was found to be a satisfactory and better blocking agent than skimmed milk. Up to 10(4.28) TCID50 poliovirus particles/0.1 ml of stool collected in 10% skimmed milk could be detected depending on the initial titer of the antigen specific capturing serum/IgG immobilized on nitrocellulose membrane. Percentage of skimmed milk in the transport medium, composition and pH of the dilution buffer and chloroform treatment of the stool specimens before the test were important determinants of the specificity of the test. Polyvinyl affinity membrane did not appear to be superior to nitrocellulose membrane as a solid phase support.
Zakir Hussain
A M
AM
Department of Tropical Medicine and Medical Microbiology, University of Hawaii at Manoa, Honolulu, 96816.
Marchette
N J
NJ
James
J F
JF
D000914:Antibodies, Viral / Q000276:immunology; D003101:Collodion; D005243:Feces / Q000382:microbiology*; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000276:immunology; D017955:Poliovirus / Q000276:immunology / Q000302:isolation & purification*
D000914:Antibodies, Viral; D007074:Immunoglobulin G; D003101:Collodion