Production of somatic extract of adult Haemonchus contortus and its use in an indirect Enzyme-linked Immunosorbent assay for detection of ovine immunoglobulin G
Abstract
One of the most limiting factors to the small ruminant husbandry is gastrointesinal verminosis, especially in mild whether regions, where Haemonchus contortus is the predominant worm. To understand sheep immune response to this parasite and to identify the more responsive animals, it is necessary to adopt adequate methods that can measure the host’s immune response. This work was conducted in order to produce a somatic extract of H.contortus and to employ it in an indirect enzyme-linked Immunosorbent assay (ELISA) to detect ovine IgG. For obtaining the antigen, about 1.000 adult worms (both male and female) were washed, macerated, submitted to freezing and thawing cycles, sonicated and centrifuged. Samples from 20 ewes naturally infected were used as positive standards. Negative standards were 15 pre-colostral samples and those from two lambs kept indoors and parasite-free. It was tested several dilutions of antigen, sera and anti- antigen concentration was 5μg.mL-1 and the optimal dilutions of sera and conjugated were
1:80 and 1:1000, respectively.
Keywords
IgG; Haemonchus contortus; ELISA
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PDF (Português (Brasil))DOI: http://dx.doi.org/10.5380/avs.v13i2.12892