Whole-blood samples were taken from the cubital vein. Blood cell counts of leukocytes, haematocrit, thrombocytes, and leukocyte subtypes were analysed using the Sysmex XT 2000 Haematology Autoanalyser (Sysmex, Kobe, Japan) according to the manufacturer’s recommendation [44 (link)]. Serum cortisol levels were measured with the Advia Centaur Cortisol Assay (Siemens Healthcare Diagnostics, Eschborn, Germany) without within-sample replication [43 (link)].
Advia centaur cortisol assay
Manufactured by Siemens
Sourced in Germany
The ADVIA Centaur Cortisol assay is a laboratory equipment product from Siemens. It is designed to measure the concentration of cortisol, a hormone, in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Multi test imk kits, by BD (1 mentions)
Cba human chemokine kit, by BD (1 mentions)
Fasc calibur, by BD (1 mentions)
Cytometric bead array cba human inflammatory cytokines kit, by BD (1 mentions)
Quantikine human bdnf immunoassay kit, by R&D Systems (1 mentions)
Vacutainer plus plastic k2edta tubes, by BD (1 mentions)
6 protocols using advia centaur cortisol assay
1
Routine Blood Analysis Protocol
2
Serum Cortisol Levels in Guelph
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Blood samples were collected from participants in the morning by Lifelabs Medical Laboratory Services in Guelph. Serum cortisol levels were also analyzed by Lifelabs using the Siemens Advia Centaur cortisol assay, a competitive immunoassay with chemiluminescent detection. Lifelabs did not require participants to fast for this test.
3
Comprehensive Cytokine and Cellular Analysis
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Cytokine and chemokine assessments. The BD™ Cytometric Bead Array (CBA) Human Inflammatory Cytokines Kit was used to quantitatively measure IL-8, IL-1β, IL-6, IL-10, TNF and IL-12P protein levels in a single sample.
BD™ CBA Human Chemokine Kit was used to quantitatively measure IL-8 (CXCL8/IL-8), RANTES (CCL5/RANTES), monokine induced by IFN-γ (CXCL9/MIG) and IFN-γ-induced protein-10 (CXCL10/IP-10) levels in a single sample. Cell subtype assessments. BD™ Multitest IMK kits were used with CD3/CD8/CD45/CD4 reagent and CD3/CD16 + CD56/CD45/ CD19 reagent. Flow cytometry analysis measures the emission of optical signals, after passing through a laser beam. In our investigation a "lyse -no wash" method was used (35) . The measurement was performed using CellQuest Protocol according to the guidelines of the manufacturer on a BD™ FascCalibur. Assessment of cortisol levels. Serum for analysis was obtained between 07.40 h and 10.15 h. Cortisol levels were analyzed using Siemens ADVIA Centaur ® cortisol assay, a competitive immunoassay using direct chemiluminescent technology.
BD™ CBA Human Chemokine Kit was used to quantitatively measure IL-8 (CXCL8/IL-8), RANTES (CCL5/RANTES), monokine induced by IFN-γ (CXCL9/MIG) and IFN-γ-induced protein-10 (CXCL10/IP-10) levels in a single sample. Cell subtype assessments. BD™ Multitest IMK kits were used with CD3/CD8/CD45/CD4 reagent and CD3/CD16 + CD56/CD45/ CD19 reagent. Flow cytometry analysis measures the emission of optical signals, after passing through a laser beam. In our investigation a "lyse -no wash" method was used (35) . The measurement was performed using CellQuest Protocol according to the guidelines of the manufacturer on a BD™ FascCalibur. Assessment of cortisol levels. Serum for analysis was obtained between 07.40 h and 10.15 h. Cortisol levels were analyzed using Siemens ADVIA Centaur ® cortisol assay, a competitive immunoassay using direct chemiluminescent technology.
4
Quantifying Stress Biomarkers in Serum
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Blood samples were taken before breakfast at 07:00 on the day after admission and were centrifuged immediately after blood was drawn and clotting confirmed. Serum samples were stored at -80℃ until the time of processing. levels of BDNF were measured using a Quantikine Human BDNF Immunoassay Kit (R&D Systems, Minneapolis, MN, USA) according to the instructions from the manufacturer and as previously described 29) . The lower limit of the detection was 20 pg/ml and the intra-assay coefficient of variation was < 8%.
Serum levels of cortisol were measured via competitive immunoassay using the ADVIA Centaur Cortisol assay (Siemens Healthcare Diagnostic Inc., Tokyo, Japan). This assay measures serum cortisol concentrations up to 75 μg/dl (2,069 nmol/l) with a minimum detectable concentration (analytical sensitivity) of 0.20 μg/dl (5.5 nmol/l). The intra-assay CVs were < 15% 30) .
Serum levels of DHEA-S were measured at the SRL Laboratory (Tokyo, Japan) using a chemiluminescent immunoassay (CLEIA; Access DHEA-S, Beckman Coulter KK, Tokyo, Japan). The lower limit of the detection for DHEA-S were 11 ng/ml. Inter-and intra-assay coefficients of variation were 10.6% and 7.2% for DHEA-S 25) .
Serum levels of cortisol were measured via competitive immunoassay using the ADVIA Centaur Cortisol assay (Siemens Healthcare Diagnostic Inc., Tokyo, Japan). This assay measures serum cortisol concentrations up to 75 μg/dl (2,069 nmol/l) with a minimum detectable concentration (analytical sensitivity) of 0.20 μg/dl (5.5 nmol/l). The intra-assay CVs were < 15% 30) .
Serum levels of DHEA-S were measured at the SRL Laboratory (Tokyo, Japan) using a chemiluminescent immunoassay (CLEIA; Access DHEA-S, Beckman Coulter KK, Tokyo, Japan). The lower limit of the detection for DHEA-S were 11 ng/ml. Inter-and intra-assay coefficients of variation were 10.6% and 7.2% for DHEA-S 25) .
5
Cortisol and NEFA Measurement Protocol
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Concentrations of cortisol in plasma were determined in a single assay according to the manufacturer’s instructions (ADVIA Centaur® Cortisol assay; Siemens Healthcare Pty Ltd, Bayswater, Victoria, Australia). Assay sensitivity was 2.0 ng/mL, and the intra-assay coefficient of variation was 3.5%. The NEFA were also determined in a single assay (Randox Australia Pty Ltd, Parramatta, Australia) previously used in cattle20 (link). Assay sensitivity was 0.03 mEq/L, and the intra-assay coefficient of variation was 1.4%.
6
Assessing HPA-axis Activation
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HPA-axis activation was assessed through plasma adrenocorticotropic hormone (ACTH) levels, determined using sandwich chemiluminescence immunoassay (LaisonXL, DiaSorin S.p.A., Saluggia, Italy), and cortisol levels, measured by chemiluminescent immunoassay (ADVIA Centaur Cortisol assay, Siemens Healthcare Diagnostics, Munich, Germany). Blood samples were collected in plastic tubes (BD Vacutainer® Plus Plastic K2-EDTA Tubes, Franklin Lakes, NJ, USA), centrifuged, and aliquoted for hormonal determinations.
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