Publication: Measurement of chromogranin A in porcine saliva: validation of a time-resolved immunofluorometric assay and evaluation of its application as a marker of acute stress
Authors
Escribano Tortosa, Damián ; Soler Vasco, Laura ; Gutiérrez Montes, Ana María ; Martínez Subiela, Silvia ; Cerón Madrigal, José Joaquín
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Cerón Madrigal, José Joaquín
Publisher
Elsevier
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DOI
https://doi.org/10.1017/S1751731112002005
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info:eu-repo/semantics/article
Description
©2013. This manuscript version is made available under the CC-BY4.0
license http://creativecommons.org/licenses/by/4.0/. This document is the Published version of a Published Work that appeared in final form in Animal. To access the final edited and published work see https://doi.org/10.1017/S1751731112002005.
Abstract
The objective of this study was to develop and validate a time-resolved immunofluorometric assay (TR-IFMA) for porcine salivary chromogranin A (CgA) measurements, using a species-specific antibody, and evaluate its behaviour in an acute stress model. Polyclonal antibodies were produced in rabbits immunized with a synthetic porcine fragment of CgA3592379 and used to develop a sandwich TR-IFMA. This TR-IFMA was analytically validated and showed intra- and inter-assay coefficients of variation of 6.23% and 5.82%, respectively, an analytical limit of detection of 4.27 3 1023 mg/ml and a limit of quantification of 24.5 3 1023 mg/ml. The assay also demonstrated a high level of accuracy, as determined by linearity under dilution ( r 5 0.975) and recovery tests. When a model of experimental acute stress, in which animals were immobilized for 3 min with a nose snare (stressor stimulus), was applied, a significant increase ( P , 0.05) in CgA levels in saliva was detected at 15 min post-stressor stimulus. These results indicate that the assay developed in this study could measure CgA in porcine saliva in a reliable way and that the concentrations of CgA in saliva samples of pigs increase after an acute stress situation.
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Citation
Animal (2013), 7:4, p.p. 640–647
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