Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure
BACKGROUND: The neuropeptide oxytocin is increasing the focus of many studies investigating human and animal social behaviours and diseases. However, interpretation and comparison of results is made difficult by a lack of consistent methodological approaches towards analysing this hormone. NEW METHO...
Published in: | Journal of Neuroscience Methods |
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Format: | Article in Journal/Newspaper |
Language: | English |
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2014
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Online Access: | https://risweb.st-andrews.ac.uk/portal/en/researchoutput/validation-of-an-enzymelinked-immunoassay-elisa-for-plasma-oxytocin-in-a-novel-mammal-species-reveals-potential-errors-induced-by-sampling-procedure(23d599b5-4f88-4237-a345-89189ed3e53e).html https://doi.org/10.1016/j.jneumeth.2014.01.019 https://research-repository.st-andrews.ac.uk/bitstream/10023/4541/3/JNM2014.pdf |
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ftunstandrewcris:oai:risweb.st-andrews.ac.uk:publications/23d599b5-4f88-4237-a345-89189ed3e53e 2023-05-15T17:58:53+02:00 Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure Robinson, Kelly Joanne Hazon, Neil Lonergan, Mike Pomeroy, Patrick 2014-01 application/pdf https://risweb.st-andrews.ac.uk/portal/en/researchoutput/validation-of-an-enzymelinked-immunoassay-elisa-for-plasma-oxytocin-in-a-novel-mammal-species-reveals-potential-errors-induced-by-sampling-procedure(23d599b5-4f88-4237-a345-89189ed3e53e).html https://doi.org/10.1016/j.jneumeth.2014.01.019 https://research-repository.st-andrews.ac.uk/bitstream/10023/4541/3/JNM2014.pdf eng eng info:eu-repo/semantics/openAccess Robinson , K J , Hazon , N , Lonergan , M & Pomeroy , P 2014 , ' Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure ' , Journal of Neuroscience Methods , vol. 226 , pp. 73-79 . https://doi.org/10.1016/j.jneumeth.2014.01.019 Enzyme-linked immunoassay (ELISA) Plasma oxytocin Raw plasma Grey seals (Halichoerus grypus) Harbour seals (Phoca vitulina) EDTA Heparin article 2014 ftunstandrewcris https://doi.org/10.1016/j.jneumeth.2014.01.019 2021-12-26T14:24:09Z BACKGROUND: The neuropeptide oxytocin is increasing the focus of many studies investigating human and animal social behaviours and diseases. However, interpretation and comparison of results is made difficult by a lack of consistent methodological approaches towards analysing this hormone. NEW METHOD: This study determined the sample collection and analysis protocols that cause the least amounts of protocol dependant variation in plasma oxytocin concentrations detected by ELISA. The effect of vacutainer type, sample extraction prior to analysis and capture and restraint protocol were investigated while validating an assay protocol for two novel species, grey seals (Halichoerus grypus) and harbour seals (Phoca vitulina). RESULTS: Where samples are extracted prior to analysis, vacutainer type (EDTA mean: 8.25±0.56pg/ml, heparin mean: 8.25±0.62pg/ml, p=0.82), time taken to obtain a sample and restraint protocol did not affect the concentration of oxytocin detected. However, concentrations of oxytocin detected in raw plasma samples were significantly higher than those in extracted samples, and varied significantly with vacutainer type (EDTA mean: 534.4±43.7pg/ml, heparin mean: 300.9±19.6pg/ml, p<0.001) and capture and restraint methodology. There was no relationship between oxytocin concentrations detected in raw and extracted plasma (p=0.25). COMPARISON WITH EXISTING METHOD(S): Over half the reviewed published studies analysing plasma oxytocin use raw plasma and different vacutainer types are used without consistency or justification through-out the literature. CONCLUSIONS: We caution that studies using raw plasma are likely to over estimate oxytocin concentrations, cannot be used to accurately infer true values via correlations and are susceptible to variation according vacutainer type. Article in Journal/Newspaper Phoca vitulina University of St Andrews: Research Portal Journal of Neuroscience Methods 226 73 79 |
institution |
Open Polar |
collection |
University of St Andrews: Research Portal |
op_collection_id |
ftunstandrewcris |
language |
English |
topic |
Enzyme-linked immunoassay (ELISA) Plasma oxytocin Raw plasma Grey seals (Halichoerus grypus) Harbour seals (Phoca vitulina) EDTA Heparin |
spellingShingle |
Enzyme-linked immunoassay (ELISA) Plasma oxytocin Raw plasma Grey seals (Halichoerus grypus) Harbour seals (Phoca vitulina) EDTA Heparin Robinson, Kelly Joanne Hazon, Neil Lonergan, Mike Pomeroy, Patrick Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure |
topic_facet |
Enzyme-linked immunoassay (ELISA) Plasma oxytocin Raw plasma Grey seals (Halichoerus grypus) Harbour seals (Phoca vitulina) EDTA Heparin |
description |
BACKGROUND: The neuropeptide oxytocin is increasing the focus of many studies investigating human and animal social behaviours and diseases. However, interpretation and comparison of results is made difficult by a lack of consistent methodological approaches towards analysing this hormone. NEW METHOD: This study determined the sample collection and analysis protocols that cause the least amounts of protocol dependant variation in plasma oxytocin concentrations detected by ELISA. The effect of vacutainer type, sample extraction prior to analysis and capture and restraint protocol were investigated while validating an assay protocol for two novel species, grey seals (Halichoerus grypus) and harbour seals (Phoca vitulina). RESULTS: Where samples are extracted prior to analysis, vacutainer type (EDTA mean: 8.25±0.56pg/ml, heparin mean: 8.25±0.62pg/ml, p=0.82), time taken to obtain a sample and restraint protocol did not affect the concentration of oxytocin detected. However, concentrations of oxytocin detected in raw plasma samples were significantly higher than those in extracted samples, and varied significantly with vacutainer type (EDTA mean: 534.4±43.7pg/ml, heparin mean: 300.9±19.6pg/ml, p<0.001) and capture and restraint methodology. There was no relationship between oxytocin concentrations detected in raw and extracted plasma (p=0.25). COMPARISON WITH EXISTING METHOD(S): Over half the reviewed published studies analysing plasma oxytocin use raw plasma and different vacutainer types are used without consistency or justification through-out the literature. CONCLUSIONS: We caution that studies using raw plasma are likely to over estimate oxytocin concentrations, cannot be used to accurately infer true values via correlations and are susceptible to variation according vacutainer type. |
format |
Article in Journal/Newspaper |
author |
Robinson, Kelly Joanne Hazon, Neil Lonergan, Mike Pomeroy, Patrick |
author_facet |
Robinson, Kelly Joanne Hazon, Neil Lonergan, Mike Pomeroy, Patrick |
author_sort |
Robinson, Kelly Joanne |
title |
Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure |
title_short |
Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure |
title_full |
Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure |
title_fullStr |
Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure |
title_full_unstemmed |
Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure |
title_sort |
validation of an enzyme-linked immunoassay (elisa) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure |
publishDate |
2014 |
url |
https://risweb.st-andrews.ac.uk/portal/en/researchoutput/validation-of-an-enzymelinked-immunoassay-elisa-for-plasma-oxytocin-in-a-novel-mammal-species-reveals-potential-errors-induced-by-sampling-procedure(23d599b5-4f88-4237-a345-89189ed3e53e).html https://doi.org/10.1016/j.jneumeth.2014.01.019 https://research-repository.st-andrews.ac.uk/bitstream/10023/4541/3/JNM2014.pdf |
genre |
Phoca vitulina |
genre_facet |
Phoca vitulina |
op_source |
Robinson , K J , Hazon , N , Lonergan , M & Pomeroy , P 2014 , ' Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure ' , Journal of Neuroscience Methods , vol. 226 , pp. 73-79 . https://doi.org/10.1016/j.jneumeth.2014.01.019 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1016/j.jneumeth.2014.01.019 |
container_title |
Journal of Neuroscience Methods |
container_volume |
226 |
container_start_page |
73 |
op_container_end_page |
79 |
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1766167612007383040 |