Development of monoclonal antibody ELISA for simultaneous detection of bovine coronavirus, rotavirus serogroup A, and Escherichia coli K99 antigen in feces of calves.
A rapid ELISA was developed for simultaneous detection of bovine coronavirus (BCV), rotavirus (RV) serogroup A, and Escherichia coli K99 antigen in feces of calves. A mixture of 3 monoclonal antibodies specific for BCV, RV, or K99 was used successfully to capture the antigens; the same antibodies labeled with peroxidase were used to detect BCV, RV, or K99. The triple ELISA was compared with standard reference diagnostic methods by examining feces from experimentally and naturally infected and healthy calves. All the components of the test were highly specific (greater than 90%) and sensitive (BCV, 77%; K99, 93%; RV, 100%) when used in a format requiring short incubation steps at 20 C and visual recording of results.
Ministry of Agriculture, Fisheries, and Food, Central Veterinary Laboratory, Weybridge, Surrey, United Kingdom.
C J
CJ
Thorns
M M
MM
Bell
D
D
Chasey
J
J
Chesham
P L
PL
Roeder
D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000954:Antigens, Surface / Q000032:analysis; D000956:Antigens, Viral / Q000032:analysis*; D001422:Bacterial Adhesion; D001427:Bacterial Toxins; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis; D002460:Cell Line; D003332:Coronaviridae / Q000276:immunology* / Q000648:ultrastructure; D003967:Diarrhea / Q000175:diagnosis / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D004926:Escherichia coli / Q000276:immunology* / Q000648:ultrastructure; D005243:Feces / Q000382:microbiology*; D008854:Microscopy, Electron; D011237:Predictive Value of Tests; D012401:Rotavirus / Q000276:immunology* / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D000956:Antigens, Viral; D001427:Bacterial Toxins; D000939:Epitopes; C031429:K99 antigen