Detection of circulating hypodermin C: an antigen capture ELISA for diagnosis of cattle grub (Diptera: Oestridae) infestations.
An antigen capture assay for the detection of circulating hypodermin C was developed for diagnosis of hypodermosis. A murine monoclonal antibody to recombinant hypodermin C was raised using rapid immunization and a one-step hybridization-cloning technique. A highly reactive, specific monoclonal antibody was tested using sera spiked with known quantities of purified, native hypodermin C or with recombinant hypodermin C. Sensitivity of 96.4% and specificity of 95.6% for the antigen capture assay was assessed using a panel of sera from animals unexposed to cattle grubs and from cattle with palpation proven cattle grub infestations. Data from this panel of sera was used to establish the cut-off OD for further testing. The kinetics of circulating hypodermin C was assessed using the assay in three groups of cattle artificially infested with 50, 100 or 200 first instar Hypoderma lineatum. Antigen was first detected approximately 6 weeks after infestation. The amount of antigen detected increased in each group of animals reaching peaks at different times in each group. Levels of antigen fell quickly following arrival of grubs at the back and completion of the molt to second instar.
Parasitología y Enfermedades Parasitarias, Departamento de Patologi;a Animal, Facultad de Veterinaria, Universidad de Santiago de Compostela, Lugo, Spain.
Rosario
R
Panadero-Fontán
Ceferino
C
Lopez-Sández
Francisco
F
Parra-Fernández
Patrocinio
P
Morrondo-Pelayo
Pablo
P
Díez-Baños
Douglas D
DD
Colwell
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000097:blood; D000941:Antigens / Q000276:immunology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000469:parasitology; D004175:Diptera / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D007000:Hypodermyiasis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D012680:Sensitivity and Specificity; D012697:Serine Endopeptidases / Q000097:blood* / Q000276:immunology; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000941:Antigens; D012697:Serine Endopeptidases; C070019:hypodermin C