Development, characterization, and diagnostic applications of monoclonal antibodies against bovine rotavirus.
Hybridomas secreting monoclonal antibodies (MAbs) against the Nebraska calf diarrhea strain of bovine rotavirus (BRV) were characterized. Indirect fluorescent-antibody assay, immunodot assay, and immunoprecipitation were used to select hybridomas that produced anti-BRV MAbs. Seven of the MAbs were shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot assay to be reactive with the BRV outer capsid protein, VP7, which has a molecular mass of 37.5 kDa. None of the seven MAbs were reactive with canine rotavirus, bovine coronavirus, or uninfected Madin-Darby bovine kidney cells. Two clones, 8B4 (immunoglobulin G2a [IgG2a]) and 2B11 (IgG1), were found suitable for use in an antigen capture enzyme-linked immunosorbent assay for detecting BRV in bovine fecal samples. Both were subtype A specific (G6 subtype) but did not react with all isolates of BRV group A.
Al-Yousif
Y
Y
Department of Diagnostic Medicine-Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, Kansas 66506, USA.
Al-Majhdi
F
F
Chard-Bergstrom
C
C
Anderson
J
J
Kapil
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D002213:Capsid / Q000276:immunology*; D036022:Capsid Proteins; D002417:Cattle; D002460:Cell Line; D004285:Dogs; D051379:Mice; D008807:Mice, Inbred BALB C; D012401:Rotavirus / Q000276:immunology*; D012400:Rotavirus Infections / Q000175:diagnosis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; C050779:VP7 protein, Rotavirus