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Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure

Research output: Contribution to journalArticle

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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.

In: Journal of Neuroscience Methods, Vol. 226, 01.2014, p. 73-79.

Research output: Contribution to journalArticle

Harvard

Robinson, KJ, 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

APA

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, 226, 73-79. https://doi.org/10.1016/j.jneumeth.2014.01.019

Vancouver

Robinson KJ, Hazon N, Lonergan M, Pomeroy P. 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. 2014 Jan;226:73-79. https://doi.org/10.1016/j.jneumeth.2014.01.019

Author

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. In: Journal of Neuroscience Methods. 2014 ; Vol. 226. pp. 73-79.

Bibtex - Download

@article{23d599b54f884237a34589189ed3e53e,
title = "Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure",
abstract = "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.",
keywords = "Enzyme-linked immunoassay (ELISA), Plasma oxytocin, Raw plasma, Grey seals (Halichoerus grypus), Harbour seals (Phoca vitulina), EDTA, Heparin",
author = "Robinson, {Kelly Joanne} and Neil Hazon and Mike Lonergan and Patrick Pomeroy",
note = "This work was conducted as part of a PhD, which was funded by the Natural Environment Research Council (NERC), UK and by SMRU Marine, St Andrews, UK.",
year = "2014",
month = "1",
doi = "10.1016/j.jneumeth.2014.01.019",
language = "English",
volume = "226",
pages = "73--79",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure

AU - Robinson, Kelly Joanne

AU - Hazon, Neil

AU - Lonergan, Mike

AU - Pomeroy, Patrick

N1 - This work was conducted as part of a PhD, which was funded by the Natural Environment Research Council (NERC), UK and by SMRU Marine, St Andrews, UK.

PY - 2014/1

Y1 - 2014/1

N2 - 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.

AB - 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.

KW - Enzyme-linked immunoassay (ELISA)

KW - Plasma oxytocin

KW - Raw plasma

KW - Grey seals (Halichoerus grypus)

KW - Harbour seals (Phoca vitulina)

KW - EDTA

KW - Heparin

U2 - 10.1016/j.jneumeth.2014.01.019

DO - 10.1016/j.jneumeth.2014.01.019

M3 - Article

VL - 226

SP - 73

EP - 79

JO - Journal of Neuroscience Methods

T2 - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

ER -

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ID: 98220849