Body temperature was assessed via medical-grade oral thermometer (WelchAllyn SureTemp Plus) at each study visit. Serum proinflammatory markers were assessed at baseline, 1, 2, and 3 days post-vaccination. At each study visit, whole blood was collected into vacutainer tubes while subjects were in a seated position. On follow-up days, blood samples for the same woman were collected within a 2 hour window of collection of the baseline sample for that particular woman to ensure that sample timepoints were approximately 24 hours apart. Samples were immediately centrifuged, aliquoted, and placed in −80°C freezer storage until analysis. Serum levels of interleukin(IL)-6, tumor necrosis factor (TNF)-α, IL-8, and IL-1β were assayed in duplicate with ultra-sensitive multiplex kits from Meso Scale Discovery (MSD) and chemilluminescence methodology using the Immulite 1000 (Siemens Healthcare Diagnostics, Inc., 1717 Deerfield Rd., Deerfield, Il.). Serum levels of macrophage migration inhibitory factor (MIF) were assayed in duplicate using ultra-sensitive multiplex kits from R&D Systems (Minneapolis, MN) per kit instructions.
Ultra sensitive multiplex kits
Manufactured by Mesoscale
Ultra-sensitive multiplex kits are laboratory equipment designed to detect and analyze multiple target analytes simultaneously with high sensitivity. The core function of these kits is to enable precise and efficient quantification of various biomolecules in a sample.
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Lab products found in correlation
2 protocols using ultra sensitive multiplex kits
1
Assessing Inflammatory Markers Post-Vaccination
2
Cytokine and CRP Measurement Protocol
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Serum levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-8, and IL-1β were assayed in duplicate with ultra-sensitive multiplex kits from Meso Scale Discovery (MSD) and chemilluminescence methodology using the Sector Imager 2400 (Meso Scale Discovery, 9238 Gaithers Rd., Gaithersburg, Md). Limits of detection were 0.61 pg/ml for IL-6, 2.4 pg/ml for TNF-α, 0.3 pg/ml for IL-8, and 0.61 pg/ml for IL-1β. All values were above the limits of detection. Intra-assay coefficients of variation (CVs) were between 5.21 to 10.66 and the inter-assay CVs were between 8.4 to 11.17. CRP values were determined using chemilluminescence methodology using the Immulite 1000 (Siemens Healthcare Diagnostics, Inc., 1717 Deerfield Rd., Deerfield, Il.) The limit of detection is 0.3 mg/L; all values were above the limit of detection. The intra and inter-assay coefficients of variation were 5.1% and 7.3%. All samples from the same participant were batched together and assayed on the same plate. Assays for the two studies were conducted at the same time and all plates were from the same lot.
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