OBJECTIVE: Interleukin-6 (IL-6) is a pro-inflammatory cytokine that is associated with the production of C-reactive protein, the acute phase response, and chronic low-grade inflammation. In this report, we describe and validate a highly sensitive enzyme-linked immunosorbent assay (ELISA) for quantifying IL-6 at low concentrations in samples of capillary whole blood collected from a simple finger stick and dried on filter paper (dried blood spots, DBS). METHODS: A commercially available ELISA for IL-6 was modified to develop a protocol for the quantification of IL-6 in DBS samples. Procedures for sample elution and incubation were optimized for precision, reliability, accuracy, and lower limit of detection using small volumes of DBS. A set of 46 matched serum/DBS samples were used to evaluate agreement between serum and DBS results. RESULTS: The protocol demonstrated acceptable levels of precision, reliability, accuracy, and agreement with serum-based results. The lower limit of detection was sufficiently low to measure levels of IL-6 associated with both chronic, low-grade inflammation, and acute increases in inflammatory activity. CONCLUSIONS: This protocol adds to the growing panel of analytes validated for quantification in DBS samples and should facilitate future research on the causes and consequences of inflammation in diverse non-clinical settings.
OBJECTIVE:Interleukin-6 (IL-6) is a pro-inflammatory cytokine that is associated with the production of C-reactive protein, the acute phase response, and chronic low-grade inflammation. In this report, we describe and validate a highly sensitive enzyme-linked immunosorbent assay (ELISA) for quantifying IL-6 at low concentrations in samples of capillary whole blood collected from a simple finger stick and dried on filter paper (dried blood spots, DBS). METHODS: A commercially available ELISA for IL-6 was modified to develop a protocol for the quantification of IL-6 in DBS samples. Procedures for sample elution and incubation were optimized for precision, reliability, accuracy, and lower limit of detection using small volumes of DBS. A set of 46 matched serum/DBS samples were used to evaluate agreement between serum and DBS results. RESULTS: The protocol demonstrated acceptable levels of precision, reliability, accuracy, and agreement with serum-based results. The lower limit of detection was sufficiently low to measure levels of IL-6 associated with both chronic, low-grade inflammation, and acute increases in inflammatory activity. CONCLUSIONS: This protocol adds to the growing panel of analytes validated for quantification in DBS samples and should facilitate future research on the causes and consequences of inflammation in diverse non-clinical settings.
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