Method for the immunodiagnosis of hypodermosis in cattle based on the use of purified hypodermin C

摘要

A description is given of an indirect ELISA for the immunodiagnosis of hypodermosis in cattle, which comprises the following stages: 1) purification of the hypodermin C from the first instar larvae of Hypodermin lineatum, by reverse phase chromatography (HPLC), 2) adsorption of the purified hypodermin C on microtitre plates, 3) blocking of the wells using a buffer containing egg albumin and Tween 20, 4) addition of the bovine sera, 5) washing each well 5 times, 6) adding an anti-bovine IgG serum conjugated with peroxidase, 7) washing each well 5 times, 8) adding the substrate of the peroxidase, 9) stopping the reaction with 3N sulphuric acid, and 10) measuring the colour developed in each well, determining the absorbance at 450 mm.