Validation, use, and disadvantages of enzyme-linked immunosorbent assay kits for detection of cortisol in channel catfish, largemouth bass, red pacu, and golden shiners
Open Access
- 24 July 2007
- journal article
- research article
- Published by Springer Science and Business Media LLC in Fish Physiology and Biochemistry
- Vol. 34 (1), 95-101
- https://doi.org/10.1007/s10695-007-9150-9
Abstract
Measurement of cortisol response is an important tool to asses stress in fisheries research. Radioimmunoassay (RIA) is a common method for the measure of cortisol in fish. Use of enzyme-linked immunosorbent assays (ELISA) to detect cortisol would eliminate health hazards, costs of handling radioisotopes, and the short stability time associated with RIA. Enzyme-linked immunosorbent assays have been used for the determination of cortisol in several fish species. However, the ELISA method of cortisol determination in fish lacks proper validation testing. We conducted validation procedures for multiple commercial cortisol ELISA kits and compared the results to RIA. The assays were tested for four species: (1) channel catfish Ictalurus punctatus, (2) largemouth bass Micropterus salmoides, (3) red pacu Piaractus brachypomus, and (4) golden shiners Notemigonus crysoleucas. We evaluated the ELISA methods against RIA, and determined that at least one kit is suitable (accuracy: mean recovery of spiked samples, 102.8%; reproducibility: interassay coefficient of variation < 10.5% for all species; precision: intra-assay coefficient of variation < 16.8% for all species; linearity: R 2 > 0.96 for all species) for the measurement of cortisol response in fish and comparative determination of stress. All of the ELISA assay results varied by more than 10% from the cortisol concentrations detected by the RIA. The high variability of the kit results indicates that commercial ELISA kits could be utilized for qualitative determination of cortisol in fish, but should be fully validated in each laboratory for each species before being used for research.Keywords
This publication has 14 references indexed in Scilit:
- Effect of photoperiod manipulation on the growth performance and stress response of juvenile red sea bream (Pagrus major)Aquaculture, 2006
- Management of Physiological Stress in Finfish AquacultureNorth American Journal of Aquaculture, 2006
- Stress in Fishes: A Diversity of Responses with Particular Reference to Changes in Circulating CorticosteroidsIntegrative and Comparative Biology, 2002
- Cortisol in teleosts: dynamics, mechanisms of action, and metabolic regulationReviews in Fish Biology and Fisheries, 1999
- Validation of a microtitre plate ELISA for measuring cortisol in fish and comparison of stress responses of rainbow trout (Oncorhynchus mykiss) and lake trout (Salvelinus namaycush)Aquaculture, 1993
- Effects of Rearing Density on the Stress Response and Growth of Rainbow TroutJournal of Aquatic Animal Health, 1992
- Changes in Peripheral Blood Leukocyte Percentages and Function of Neutrophils in Stressed Channel CatfishJournal of Aquatic Animal Health, 1991
- Changes in Numbers of Leukocytes in Immune Organs of Juvenile Coho Salmon after Acute Stress or Cortisol TreatmentJournal of Aquatic Animal Health, 1990
- Communications: Validation of a Solid-Phase Enzyme Immunoassay Technique for the Measure of Plasma Cortisol in Rainbow TroutJournal of Aquatic Animal Health, 1990
- Serum Cortisol Levels in Channel Catfish, from Production PondsThe Progressive Fish-Culturist, 1985