Enzyme-Linked Immunoassay-Based Quantitative Measurement of Apolipoprotein B (ApoB) in Dried Blood Spots, a Biomarker of Cardiovascular Disease Risk
Enzyme-Linked Immunoassay-Based Quantitative Measurement of Apolipoprotein B (ApoB) in Dried Blood Spots, a Biomarker of Cardiovascular Disease Risk
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DOI：
10.1080/19485565.2017.1283582
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发表时间：
2017-01-01
影响因子：
1.3
通讯作者：
Snodgrass, J. Josh
中科院分区：
文献类型：
--
作者：
Eick, Geeta N.;Kowal, Paul;Snodgrass, J. Josh
载脂蛋白B（Apo B）是心血管疾病的一个强有力的预测因子，心血管疾病仍然是高收入和低收入国家死亡率的主要原因。在这里，我们采用了酶联免疫吸附试验（ELISA）开发试剂盒，用于定量测定血清和血浆中的ApoB水平，并与干血斑（DBS）一起使用。在确认DBS测定的稀释线性后，我们测量了208份静脉DBS样本中的ApoB。然后，使用Passing-Bablok回归分析和斯皮尔曼等级相关分析，我们评价了来自40名个体的匹配血浆和手指穿刺DBS样本之间ApoB值的对应性，这些个体的ApoB值涵盖了在208份vDBS样本中观察到的ApoB值范围。我们还评价了测定精密度和回收率、红细胞压积、冻融循环次数和不同储存温度对DBS中ApoB水平的影响。血浆和DBS ApoB水平之间存在强烈的显著相关性，偏倚很小。测定精密度和回收率在美国政府生物分析测定验证行业指南推荐的范围内。测定不受DBS基质或生理红细胞压积水平的影响。这种基于DBS的ELISA检测将有助于在以前未探索的人群中进行人群规模的心血管风险评估。
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 ELISA assay will facilitate population-scale assessment of cardiovascular risk in previously unexplored populations.