Validation of an enzyme-linked immunosorbent assay developed for measuring cortisol concentration in human saliva and serum for its applicability to analyze cortisol in pig saliva

BackgroundThe purpose of this study was to validate a commercially available enzyme-linked immunosorbent assay (ELISA) developed for measuring free cortisol in human saliva and total cortisol concentration in diluted human serum, for its applicability in measuring cortisol concentration in pig saliva. Collection of saliva is less stressful than e.g. blood sampling, and is a non-invasive method.FindingsSaliva was collected by allowing sows to chew on cotton swabs held by forceps. Thereafter, the swabs were centrifuged to retrieve the saliva. The ELISA was performed according to instructions provided by the manufacturer. To validate the ELISA, determination of the intra-assay coefficient of variation (CV), inter-assay CV, recovery, linearity and parallelism was performed. The intra-assay CV was below 10% and inter-assay CV below 15% for samples of high, medium and low cortisol concentrations. The mean recovery was 117% and the linearity and parallelism showed an r2-value of 0.994 and 0.993, respectively. For biological assessment of induced social stress, two saliva samples were collected in the morning from 6 primiparous and 21 multiparous sows. One sample was collected when the sows were individually housed in a farrowing pen and a second sample was collected when the sows were group housed. The primiparous sows had a significant higher cortisol concentration compared to the multiparous sows when group housed.ConclusionThe results obtained in this validation study indicate that the ELISA is suitable for measuring cortisol concentration in porcine saliva.

• Publication year

  2014

• Venue

  Acta Veterinaria Scandinavica

• Publication date

  2014-09-06

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1186/s13028-014-0055-1PMID 25193181PMCID 4172964
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Health and immune traits of Basque and Large White pigs housed in a conventional or enriched environment.

  2012cited by this paper
• Sorting by parity to reduce aggression toward first-parity sows in group-gestation housing systems.

  2012cited by this paper
• Validation and quality control of ELISAs for the use with human saliva samples.

  2012cited by this paper
• Validation of an automated chemiluminescent immunoassay for salivary cortisol measurements in pigs

  2012cited by this paper
• Influence of housing and social changes on growth, behaviour and cortisol in piglets at weaning.

  2012cited by this paper
• Validation of an Automated Method for Salivary Alpha-Amylase Measurements in Pigs (Sus Scrofa Domesticus) and its Application as a Stress Biomarker

  2011cited by this paper
• Comparison of salivary cortisol and calculated free plasma cortisol during low-dose ACTH test in healthy subjects.

  2010cited by this paper
• Effects of continuous elevated cortisol concentrations during oestrus on concentrations and patterns of progesterone, oestradiol and LH in the sow.

  2009cited by this paper
• The effect of management strategies and parity on the behaviour and physiology of gestating sows housed in an electronic sow feeding system

  2008cited by this paper
• Pig's responses to repeated social regrouping and relocation during the growing-finishing period

  2007cited by this paper
• Impact of ACTH during oestrus on the ultrastructure of the spermatozoa and their environment in the tubal reservoir of the postovulatory sow.

  2006cited by this paper
• Influence of repeated regrouping on reproduction in gilts.

  2006cited by this paper
• Validation of a behavioral observation tool to assess pig welfare.

  2006cited by this paper
• Effects of floor-feeding and the presence of a foraging substrate on the behaviour and stress physiological response of individually housed gilts

  2003cited by this paper
• Hormones as indicators of stress.

  2002cited by this paper
• Urinary cortisol as an additional tool to assess the welfare of pregnant sows kept in two types of housing.

  2002cited by this paper
• Radioimmunoassay for Cortisol in Pig Saliva and Serum

  1997cited by this paper
• The circadian rhythm of cortisol in the saliva of young pigs.

  1996cited by this paper
• The Specific-Stress-Free housing system has positive effects on productivity, health, and welfare of pigs.

  1995cited by this paper
• Changes in pig salivary cortisol in response to transport simulation, food and water deprivation, and mixing.

  1989cited by this paper

• Development of a Duplex qPCR Assay for Detecting Porcine Adulteration in Gudeg Krecek: A Complex Traditional Indonesian Food Matrix

  2026cites this paper
• Development and application of truncated cap protein-based indirect ELISA for screening antibodies against porcine circovirus 3.

  2026cites this paper
• Balancing Objectivity and Welfare: Physiological and Behavioural Responses of Guide Dogs During an Independent Certification Protocol

  2025cites this paper
• Phenotypic assessment of F0 generation lactating sows genomically selected for heat stress tolerance.

  2025cites this paper
• An In-House ELISA for Anti-Porcine Circovirus Type 2d (PCV2d) IgG: Analytical Validation and Serological Correlation

  2025cites this paper
• Investigating the development of piglet feeding behaviour during the immediate postweaning phase using computer vision.

  2025cites this paper
• Blood Sampling in Göttingen Minipigs—A Case Study of Two Standard Methods and Clicker Training as a Restraint-Free Alternative

  2025cites this paper
• Assessing salivary cortisol and testosterone as non-invasive biomarkers for GnRH-immunocastration efficiency in heavy pigs

  2025cites this paper
• Impact of farrowing crate enrichment strategies on the welfare and performance of sows, suckling piglets, and post-weaned piglets

  2025cites this paper
• An Ego-Involving Motivational Climate Can Trigger Inflammation, a Threat Appraisal, and Basic Psychological Need Frustration in an Achievement Context.

  2024influential citation
• Light source-free smartphone detection of salivary cortisol via colorimetric lateral flow immunoassay using a photoluminescent film.

  2024cites this paper
• RELIABILITY OF AMYLOID DETECTION IMMUNOASSAY KITS FOR ALZHEIMER’S DISEASE SCREENING: A PRELIMINARY STUDY

  2024cites this paper
• Clicker Training in Minipigs to Reduce Stress during Blood Collection—An Example of Applied Refinement

  2024cites this paper
• Exposure to natural stray currents of low voltage affects the behaviour and some stress biomarkers of weaned piglets

  2024cites this paper
• Recent Advances in Self‐Powered Electrochemical Biosensors for Early Diagnosis of Diseases

  2024cites this paper
• Ultrasensitive electrochemical sensor for detection of salivary cortisol in stress conditions

  2024cites this paper
• Plasma procalcitonin kinetics in healthy dogs and dogs undergoing tibial plateau leveling osteotomy.

  2023cites this paper
• Effect of co-mingling non-littermates during lactation and feed familiarity at weaning on the performance, skin lesions and health of piglet

  2023cites this paper
• Development of a cost-effective quantitative in-house ELISA assay for screening anti-S1 IgG antibodies targeting SARS-CoV-2

  2023cites this paper
• Chronic exposure to multiple stressors alters the salivary proteome of piglets

  2023cites this paper
• A trait or a state - how consistent are tail biting phenotypes in pigs?

  2023influential citation
• Basics for the potential use of saliva to evaluate stress, inflammation, immune system, and redox homeostasis in pigs

  2022cites this paper
• Prophylactic action of ayahuasca in a non-human primate model of depressive-like behavior

  2022cites this paper
• Inter-batch variability of hemagglutinin content transiently expressed in Nicotiana benthamiana plants grown under sole-source lighting and sunlight conditions before gene transfer

  2022cites this paper
• Development and evaluation of indirect antibody ELISA assay for early diagnosis and surveillance of Crimean-Congo hemorrhagic fever infection in humans.

  2022cites this paper
• Non-Invasive Antibody Assessment in Saliva to Determine SARS-CoV-2 Exposure in Young Children

  2021cites this paper
• Development of an Electrochemical CCL17/TARC Biosensor toward Rapid Triage and Monitoring of Classic Hodgkin Lymphoma.

  2021cites this paper
• Decreased IgG Antibody Response to Viral Protein Mimotopes of Oncogenic Merkel Cell Polyomavirus in Sera From Healthy Elderly Subjects

  2021cites this paper
• UPLC–TOF–MS Method for Simultaneous Quantification of Steroid Hormones in Tissue Homogenates of Zebrafish with Solid-Phase Extraction

  2021cites this paper
• Development and validation of an enzyme-linked immunosorbent assay for the determination of bovine lactoferrin in various milk products

  2021cites this paper
• Development of a peptide aptamer pair-linked rapid fluorescent diagnostic system for Zika virus detection.

  2021cites this paper
• Achievement goal theory-based psychological skills training session buffers youth athletes’ psychophysiological responses to performance stress

  2020cites this paper
• Distance-based paper/PMMA integrated ELISA-chip for quantitative detection of immunoglobulin G.

  2020cites this paper
• Development of monoclonal antibodies and simplified sandwich ELISA for canine neutrophil gelatinase-associated lipocalin A detection

  2020cites this paper
• Sensitive detection of influenza a virus based on a CdSe/CdS/ZnS quantum dot-linked rapid fluorescent immunochromatographic test.

  2020cites this paper
• Pentraxin-3 is not related to disease severity in cirrhosis and hepatocellular carcinoma patients

  2020cites this paper
• The Microbiome–Metabolome Response in the Colon of Piglets Under the Status of Weaning Stress

  2020cites this paper
• Enantioselective mixture toxicity of the azole fungicide imazalil with the insecticide α-cypermethrin in Chironomus riparius: Investigating the importance of toxicokinetics and enzyme interactions.

  2019cites this paper
• Crude fiber modulates the fecal microbiome and steroid hormones in pregnant Meishan sows.

  2019cites this paper
• Trends in Analysis of Cortisol and Its Derivatives.

  2019cites this paper
• Influence of milk pre-treatment, and changes in cheese structure and carbon dioxide solubility on the development of split defects in cheese

  2019cites this paper
• Hair or salivary cortisol analysis to identify chronic stress in piglets?

  2019cites this paper
• A sensitive and selective ELISA methodology quantifies a demyelination marker in experimental and clinical samples.

  2018cites this paper
• Effect of milk centrifugation and incorporation of high-heat-treated centrifugate on the composition, texture, and ripening characteristics of Maasdam cheese.

  2018cites this paper
• Development of enzyme-linked immunosorbent assay for early pregnancy diagnosis in cattle.

  2018cites this paper
• Human serum folate can be measured using folate binding protein linked to enzyme-labeled protein ligand binding assay (ELPLBA) as well as ELISA

  2018cites this paper
• Enhancing cognitive function of healthy Wistar rats with aqueous extract of Centella asiatica

  2018cites this paper
• Development of a smartphone-based rapid dual fluorescent diagnostic system for the simultaneous detection of influenza A and H5 subtype in avian influenza A-infected patients

  2018cites this paper
• Rapid detection of avian influenza A virus by immunochromatographic test using a novel fluorescent dye.

  2017cites this paper
• Rapid Detection of Avian Influenza Virus by Fluorescent Diagnostic Assay using an Epitope-Derived Peptide

  2017cites this paper
• Salivary alpha amylase in on-call from home fire and emergency service personnel

  2017cites this paper
• C2K77 ELISA detects cleavage of type II collagen by cathepsin K in equine articular cartilage.

  2017cites this paper
• Pyrrolidin-2-one derivatives may reduce body weight in rats with diet-induced obesity.

  2016cites this paper
• Effect of a Novel Curcumin Formulation on Adaptogenic and Endogenous Anti-oxidant /Oxidative Stress in Chronic Mild Unpredictable Stress Model in Rats

  2016cites this paper
• Seasonal variation in harbour seal (Phoca vitulina) blubber cortisol - A novel indicator of physiological state?

  2016cites this paper
• Development and validation of an enzyme linked immunosorbent assay for fluoroquinolones in animal feeds

  2015cites this paper
• Calculations for Adjusting Endogenous Biomarker Levels During Analytical Recovery Assessments for Ligand-Binding Assay Bioanalytical Method Validation

  2015cites this paper
• Aggression and cortisol levels in three different group housing routines for lactating sows

  2015cites this paper
• A Comparison of the Anorectic Effect and Safety of the Alpha2-Adrenoceptor Ligands Guanfacine and Yohimbine in Rats with Diet-Induced Obesity

  2015cites this paper
• The application of cardiac biomarkers as detectors for cardiac disease in horses

  2015cites this paper