Validation,use, and disadvantages of enzyme-linked immunosorbent assay kits for detection of cortisol in channel catfish,largemouth bass,red pacu,and golden shiners |
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Authors: | T D Sink R T Lochmann K A Fecteau |
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Institution: | (1) Department of Aquaculture and Fisheries, The University of Arkansas at Pine Bluff, 1200 N. University Dr, Mail Slot 4912, Pine Bluff, AR 71601, USA;(2) Clinical Endocrinology Service, Department of Comparative Medicine, The University of Tennessee, 2407 River Drive, VTH A205, Knoxville, TN 37996, USA |
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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. |
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Keywords: | Cortisol ELISA Validation |
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