A new immune complex dot assay for detection of rotavirus antigen in faeces.
A new immune complex dot assay (ICDA) using immune gold/silver staining is described for the sensitive and rapid detection of rotavirus in cell culture and stool specimens. The method involves spotting preformed antigen-antibody complexes onto nitrocellulose paper, followed by incubation with colloidal gold-labelled secondary antibody and silver enhancement. ICDA was sensitive and specific and detected rotavirus antigens over a wide range of concentrations. It was more sensitive than a conventional immunodot assay (CIDA) and two commercial enzyme immunoassays (EIA) based on testing serial dilutions of a positive stool specimen. Of 26 stool specimens tested ICDA detected rotavirus antigen in 17; 14 were positive by Pathfinder Rotavirus EIA, 16 by Testpack Rotavirus EIA, and direct electron microscopy (DEM) detected only 12. The ICDA offers improved sensitivity over commercial EIAs and DEM.
Wu
B R
BR
Department of Virology, Inner Mongolia Anti-Epidemic Station Huhehote, People's Republic of China.
Mahony
J B
JB
Chernesky
M A
MA
D000936:Antigen-Antibody Complex; D000956:Antigens, Viral / Q000032:analysis; D002478:Cells, Cultured; D003101:Collodion; D005243:Feces / Q000382:microbiology*; D005759:Gastroenteritis / Q000382:microbiology; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D012401:Rotavirus / Q000276:immunology / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D003101:Collodion