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| Format: | Artículo científico |
| Language: | en |
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Universidad de Córdoba
2007
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| Online Access: | https://www.redalyc.org/articulo.oa?id=69312102 |
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| _version_ | 1866815650078392320 |
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| author | Yohana Velasco-Santamaría |
| author_facet | Yohana Velasco-Santamaría |
| contents | Methodology for determination of plasma cortisol in fish using competitive enzyme-linked immunosorbent assay (ELISA) Yohana Velasco-Santamaría Pablo Cruz-Casallas Veterinaria fish ELISA plasma Cortisol rainbow trout Objective. To determine plasma cortisol procedure in fish using competitive enzymelinkedimmunosorbent assay (ELISA). Materials and methods. Two plasma samplesof juveniles rainbow trout Oncorhynchus mykiss were analized by using ELISA humankit for cortisol assay. For standard curve calibration seven standard solutions ofcortisol in human plasma (0, 20, 50, 100, 200, 400 and 800 ng.ml-1) were used. Forthe recovery test 50, 100 and 200 ng.ml-1 standard solutions of cortisol were used;for the linearity test four dilutions of the fish plasma samples (1/2, 1/4, 1/8 and 1/16)were prepared. To each well fish plasma samples and standard solutions were addedand its content were conjugated to peroxidase and subsequently enzyme substratewas added. The enzymatic reaction was stopped by addition of phosphoric acid 0.5 Mand the absorbance was read at 450 nm. The accuracy of the pipetting procedurewas assessed previously. The recovery and linearity percentages, the standard curveand parallelism were determined. Results. The standard curve showed a highcorrelation coefficient (r2 = 0.998). The cortisol concentration of two samplesfluctuated between 64 and 72 ng.ml-1. Only the 200 ng.ml-1 standard solution showeda recovery percentage superior to 80%; in contrast, in 50 and 100 ng.ml-1 therecovery percentage fluctuated between 52 and 71%. In the dilution of 1/2 to 1/8were observed a good linearity (86 to 168%); the samples showed parallelism withthe standard curve. Conclusions. The use of human plasma cortisol for ELISAprocedure is an accuracy and efficiency test for fish cortisol plasma determination. 2007 artículo científico 0122-0268 https://www.redalyc.org/articulo.oa?id=69312102 en http://www.redalyc.org/revista.oa?id=693 Revista MVZ Córdoba application/pdf Universidad de Córdoba Revista MVZ Córdoba (Colombia) Num.1 Vol.12 |
| format | Artículo científico |
| id | redalyc_69312102 |
| language | en |
| publishDate | 2007 |
| publisher | Universidad de Córdoba |
| spellingShingle | Methodology for determination of plasma cortisol in fish using competitive enzyme-linked immunosorbent assay (ELISA) Yohana Velasco-Santamaría Veterinaria fish ELISA plasma Cortisol rainbow trout Methodology for determination of plasma cortisol in fish using competitive enzyme-linked immunosorbent assay (ELISA) Yohana Velasco-Santamaría Pablo Cruz-Casallas Veterinaria fish ELISA plasma Cortisol rainbow trout Objective. To determine plasma cortisol procedure in fish using competitive enzymelinkedimmunosorbent assay (ELISA). Materials and methods. Two plasma samplesof juveniles rainbow trout Oncorhynchus mykiss were analized by using ELISA humankit for cortisol assay. For standard curve calibration seven standard solutions ofcortisol in human plasma (0, 20, 50, 100, 200, 400 and 800 ng.ml-1) were used. Forthe recovery test 50, 100 and 200 ng.ml-1 standard solutions of cortisol were used;for the linearity test four dilutions of the fish plasma samples (1/2, 1/4, 1/8 and 1/16)were prepared. To each well fish plasma samples and standard solutions were addedand its content were conjugated to peroxidase and subsequently enzyme substratewas added. The enzymatic reaction was stopped by addition of phosphoric acid 0.5 Mand the absorbance was read at 450 nm. The accuracy of the pipetting procedurewas assessed previously. The recovery and linearity percentages, the standard curveand parallelism were determined. Results. The standard curve showed a highcorrelation coefficient (r2 = 0.998). The cortisol concentration of two samplesfluctuated between 64 and 72 ng.ml-1. Only the 200 ng.ml-1 standard solution showeda recovery percentage superior to 80%; in contrast, in 50 and 100 ng.ml-1 therecovery percentage fluctuated between 52 and 71%. In the dilution of 1/2 to 1/8were observed a good linearity (86 to 168%); the samples showed parallelism withthe standard curve. Conclusions. The use of human plasma cortisol for ELISAprocedure is an accuracy and efficiency test for fish cortisol plasma determination. 2007 artículo científico 0122-0268 https://www.redalyc.org/articulo.oa?id=69312102 en http://www.redalyc.org/revista.oa?id=693 Revista MVZ Córdoba application/pdf Universidad de Córdoba Revista MVZ Córdoba (Colombia) Num.1 Vol.12 |
| title | Methodology for determination of plasma cortisol in fish using competitive enzyme-linked immunosorbent assay (ELISA) |
| topic | Veterinaria fish ELISA plasma Cortisol rainbow trout |
| url | https://www.redalyc.org/articulo.oa?id=69312102 |