B. Mourot et Py. Lebail, ENZYME-LINKED IMMUNOSORBENT-ASSAY (ELISA) FOR RAINBOW-TROUT (ONCORHYNCHUS-MYKISS) VITELLOGENIN, Journal of immunoassay, 16(4), 1995, pp. 365-377
1. A specific and simple enzyme-linked immunoassay for rainbow trout (
Onchorynchus mykiss) vitellogenin (Vtg) is described. This assay is pe
rformed using a rabbit antiserum for Vtg purified from trout plasma. 2
. This assay is based upon the competition between soluble Vtg and Vtg
adsorbed on microtiter plates, for the rabbit anti-Vtg antibody bindi
ng sites. 3. The adsorbed Vtg-antibody complexes are revealed through
the peroxidase-antiperoxidase antibody, which is colored by o-phenylen
diamin. This assay can be performed in a day and a night. 4. Under our
conditions, 90-20% of binding gave a sensibility range of 33-1473 ng/
ml. With almost a 50% binding yeld (335 ng/ml) the intra-assay coeffic
ient of variation(CV) was 5.2% (n=26) and the inter-assay CV was 12.5%
(n=5). 5. There was low immunological cross-reactivity with sera from
other salmonids and with ovary extracts. Extracts of liver from oestr
ogenized male rainbow trout yielded displacements parallel to the vite
llogenin standard and to mature female serum or oestrogenized male ser
um. 6. This enzyme immunoassay is simple and easy to use. Its great sp
ecificity allows its use only for the rainbow trout species.