title: An antigen-capture enzyme-linked immunosorbent assay (ELISA) to detect isometamidium chloride in Oncorhynchus spp.
abstract: An antigen-capture enzyme-linked immunosorbent assay (ELISA) was developed to detect and measure isometamidium chloride in the plasma of Oncorhynchus tshawytscha and O. mykiss. Isometamidium-ovalbumin conjugate and anti-isometamidium antibodies were used to coat polystyrene plates. The peroxidase saturation technique was used to optimize the coating antigen concentration; it demonstrated low affinity of the isometamidium-ovalbumin conjugate but high affinity of the anti-isometamidium antibodies for polystyrene surface sites. The optimal conditions of antiisometamidium antibodies to coat plates was at pH 7.3 and a 1:1000 dilution (0.0012 mg ml(-1) protein). The ELISA was sensitive as it detected 0.0006 mg ml(-1) of isometamidium in fish plasma. Isometamidium diluted with saline could not be detected at concentrations less than 0.05 mg ml(-1). The results indicate that this ELISA is much more sensitive when isometamidium is bound to plasma than unbound isometamidium in saline.
authors.affiliation: Department of Zoology, University of Guelph, Ontario, Canada.
authors.forename: B F
authors.initials: BF
authors.lastname: Ardelli
authors.forename: P T
authors.initials: PT
authors.lastname: Woo
mesh_terms: D000818:Animals; D002848:Chromatography, DEAE-Cellulose / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005393:Fish Diseases / Q000097:blood / Q000188:drug therapy*; D007114:Immunization / Q000662:veterinary; D007122:Immunoelectrophoresis / Q000662:veterinary; D017686:Oncorhynchus mykiss; D010047:Ovalbumin / Q000737:chemistry; D010617:Phenanthridines / Q000097:blood* / Q000627:therapeutic use; D012474:Salmon; D012680:Sensitivity and Specificity; D013954:Thyroglobulin / Q000737:chemistry; D014344:Trypanocidal Agents / Q000097:blood* / Q000627:therapeutic use
chemical_list: D010617:Phenanthridines; D014344:Trypanocidal Agents; C000702:isometamidium chloride; D010047:Ovalbumin; D013954:Thyroglobulin