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A highly sensitive multiplex immunoassay for inflammatory cytokines in dried blood spots
Thomas W McDade1,2, Aaron Miller1, Tina T Tran2
1Department of Anthropology, Northwestern University, Evanston, Illinois, USA.
Insights
A new assay allows for sensitive measurement of inflammatory cytokines (IL6, IL10, TNFα) using finger-prick dried blood spots, offering a simpler alternative to venous blood draws for studying chronic inflammation.
Area of Science:
- Biomedical Science
- Immunology
- Biochemistry
Background:
- Chronic, low-grade inflammation is linked to various health issues.
- Current methods for measuring inflammatory cytokines require venous blood, limiting accessibility.
- Quantifying low circulating cytokine concentrations is crucial for understanding chronic inflammation.
Purpose of the Study:
- To develop and validate a sensitive multiplex immunoassay protocol for measuring inflammatory cytokines in dried blood spot (DBS) samples.
- To assess the feasibility of using finger-prick DBS for quantifying specific inflammatory markers.
Main Methods:
- Utilized a multiplex electrochemiluminescent immunoassay platform.
- Evaluated assay performance including reliability, precision, lower limit of detection, and linearity of dilution.
- Compared results from DBS samples with matched plasma samples.
Main Results:
- The assay demonstrated acceptable precision and reliability for IL6, IL8, IL10, and TNFα.
- Achieved low limits of detection for all measured cytokines (<0.5 pg/ml for IL6, IL8, IL10; <1.0 pg/ml for TNFα).
- High agreement was observed between DBS and plasma samples for IL6, IL10, and TNFα, with a noted interference for IL8.
Conclusions:
- Finger-prick DBS sampling is a viable alternative to venipuncture for quantifying IL6, IL10, and TNFα in chronic inflammation research.
- Potential interference from red blood cells may affect IL8 quantification in DBS.
- This method facilitates blood collection in non-clinical settings, advancing research on immune function and health.
Objectives:
Inflammatory cytokines are key regulators of inflammation, but current measurement approaches require venous blood to quantify low circulating concentrations associated with chronic, low-grade inflammation. This article describes a highly sensitive multiplex immunoassay protocol for the measurement of IL6, IL8, IL10, and TNFα in finger stick dried blood spot (DBS) samples.
Methods:
The protocol uses a multiplex electrochemiluminescent immunoassay platform. The following measures of assay performance were evaluated: reliability (inter-assay percent coefficient of variation; %CV), precision (intra-assay %CV), lower limit of detection (LLD), linearity of dilution, and agreement with results from matched plasma samples.
Results:
Analysis of three control samples across the assay range indicated an acceptable level of precision and reliability for each cytokine. Linearity of dilution returned average values that ranged from 104.1 to 127.6% of expected. Lower limits of detection for IL6, IL8, and IL10 were <0.5, and <1.0 pg/ml for TNFα. Level of agreement in results between matched DBS and plasma samples was high for all cytokines except for IL8.
Conclusions:
Finger stick DBS sampling provides a viable alternative to venipuncture for the quantification of IL6, IL10, and TNFα at low concentrations associated with chronic inflammation. The presence of red blood cells may interfere with the quantification of IL8 in DBS. In facilitating blood collection in nonclinical settings this method can advance scientific understandings of how social and ecological contexts shape immune function and health over the life course.
