Enzyme-Linked Immunoassay-Based Quantitative Measurement of Apolipoprotein B (ApoB) in Dried Blood Spots, a Biomarker of Cardiovascular Disease Risk

Biodemography and Social Biology
Geeta EickJ Josh Snodgrass

Abstract

Apolipoprotein B (ApoB) is a strong predictor of cardiovascular disease, which remains the leading cause of mortality in both higher and lower income countries. Here, we adapted an enzyme-linked immunosorbent assay (ELISA) development kit for quantitative determination of ApoB levels in serum and plasma for use with dried blood spots (DBS). After confirming the dilution linearity of the assay for DBS, we measured ApoB in 208 venous DBS samples. Then, using Passing-Bablok regression analysis and Spearman rank correlation analysis, we evaluated the correspondence in ApoB values between matched plasma and finger-prick DBS samples from 40 individuals who had ApoB values spanning the range of ApoB values observed in the 208 vDBS samples. We also evaluated assay precision and recovery, the effects of hematocrit, number of freeze-thaw cycles, and different storage temperatures on ApoB levels in DBS. There was a strong, significant correlation between plasma and DBS ApoB levels with little bias. Assay precision and recovery were within the range recommended by the U.S. government's industry guidelines for bioanalytical assay validation. The assay was not affected by the DBS matrix or physiological hematocrit levels. This DBS-based ELIS...Continue Reading

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Citations

Mar 22, 2019·American Journal of Human Biology : the Official Journal of the Human Biology Council·Geeta N EickJ Josh Snodgrass
Feb 6, 2020·American Journal of Human Biology : the Official Journal of the Human Biology Council·Geeta N EickJ Josh Snodgrass
Aug 11, 2021·American Journal of Human Biology : the Official Journal of the Human Biology Council·Alicia M DeLouizeJ Josh Snodgrass

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Methods Mentioned

BETA
enzyme-linked immunosorbent assay
ELISA

Software Mentioned

Graphpad Prism
ACOMED Statistik
BioTek
Gen5

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