Detection of foot-and-mouth disease virus antibodies. I. "Passive" hemagglutination test.
A passive hemagglutination test has been developed to detect and measure foot-and-mouth disease virus (FMDV) antibody by using glutaraldehyde as a coupling reagent. An optimal concentration of 10 to 40 mug of virus per ml with 0.25% glutaraldehyde at 25 C for 1 hr was established for the sensitization of sheep erythrocytes. A reaction time of 18 hr at 4 C or 2 hr at 37 C induced good agglutination in the presence of specific antibody. Sensitization was carried out in phosphate buffer, whereas agglutination and preadsorption of nonspecific agglutinins from sera were performed in gelatin (0.1%, w/v)-stabilized, phosphate-buffered saline. An optimal pH of 7.2 was also established for all reactions. Antibodies derived from guinea pigs hyperimmunized by infecting with FMDV, types A, O, and C were both virus-and type-specific. Preliminary experiments showed that strain A-119 and strain A-24 Cruzeiro could also be distinguished by hemagglutination. Parallel hemagglutination and complement-fixation tests showed the former to be two to four times more sensitive than the latter.
D000447:Aldehydes; D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000918:Antibody Specificity; D005537:Aphthovirus / Q000276:immunology*; D002417:Cattle; D003168:Complement Fixation Tests; D003937:Diagnosis, Differential; D004912:Erythrocytes / Q000276:immunology; D005536:Foot-and-Mouth Disease / Q000175:diagnosis; D005780:Gelatin; D006168:Guinea Pigs; D006385:Hemagglutination Inhibition Tests; D006386:Hemagglutination Tests; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D005779:Immunodiffusion; D007202:Indicators and Reagents; D008722:Methods; D012756:Sheep; D013552:Swine; D013696:Temperature
D000447:Aldehydes; D000906:Antibodies; D007106:Immune Sera; D007202:Indicators and Reagents; D005780:Gelatin