Development and validation of chemiluminescent immunoassay for vitellogenin in five salmonid species

A highly sensitive and specific chemiluminescent immunoassay (CLIA) was dev eloped for quantification of vitellogenin (Vg) in five salmonids. The CLIA for salmon Vg was performed using the two-site method, with anti-masu salmo n beta ' -component as primary antibody and chemiluminescent acridinium-lab eled anti-rainbow trout lipovitellin F(ab)' (2) as the second antibody. Usi ng cutthroat trout Vg as the standard, the working range of the CLIA was fr om 60 pg to 500 ng Vg/ml. Intra- and inter-assay coefficients of variation ranged from 3.04 to 6.67% and 3.23 to 5.86%, respectively. For the various salmonid species, serially diluted samples of serum from vitellogenic fish ran parallel to their purified Vg standard curve in the CLIA. In male cutth roat trout maturing during the 4 months before spawning, serum Vg levels ra nged from 1.56 to 8000 ng/ml. High levels of Vg in some individuals may hav e resulted from temporary elevation of estradiol-17 beta levels in the same fish during December or January (1-2 months before spawning). This is the first report on changes in serum Vg levels in maturing male trout using CLI A, the most sensitive assay for Vg yet developed. (C) 2001 Elsevier Science Inc. All rights reserved.