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Detection of circulating hypodermin C: an antigen capture ELISA for diagnosis of cattle grub (Diptera: Oestridae) infestations
Authors:Panadero-Fontán Rosario  Lopez-Sández Ceferino  Parra-Fernández Francisco  Morrondo-Pelayo Patrocinio  Díez-Baños Pablo  Colwell Douglas D
Affiliation:Parasitología y Enfermedades Parasitarias, Departamento de Patologi;a Animal, Facultad de Veterinaria, Universidad de Santiago de Compostela, Lugo, Spain.
Abstract: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.
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