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 ELISA assay will facilitate population-scale assessment of cardiovascular risk in previously unexplored populations.

抽象的

载脂蛋白B（ApoB）是心血管疾病的有力预测指标，在高收入和低收入国家，心血管疾病仍然是导致死亡的主要原因。在这里，我们采用了一种酶联免疫吸附测定（ELISA）开发套件，用于定量测定血清和血浆中的ApoB水平，以用于干血斑（DBS）。在确认了DBS分析的稀释线性之后，我们在208个静脉DBS样品中测量了ApoB。然后，使用Passing-Bablok回归分析和Spearman秩相关分析，我们评估了来自40个个体的匹配血浆和手指刺DBS样品之间的ApoB值之间的对应性，这些个体的ApoB值跨越了208个vDBS样品中观察到的ApoB值范围。我们还评估了测定的准确性和回收率，血细胞比容的影响，冻融循环的次数，以及DBS中ApoB级别的不同存储温度。血浆和DBS ApoB水平之间存在很强的显着相关，几乎没有偏差。测定精度和回收率在美国政府针对生物分析测定验证的行业指南建议的范围内。该测定不受DBS基质或生理性血细胞比容水平的影响。这种基于DBS的ELISA测定法将有助于对以前未开发人群的心血管风险进行人群规模评估。该测定不受DBS基质或生理性血细胞比容水平的影响。这种基于DBS的ELISA测定法将有助于对以前未开发人群的心血管风险进行人群规模评估。该测定不受DBS基质或生理性血细胞比容水平的影响。这种基于DBS的ELISA测定法将有助于对以前未开发人群的心血管风险进行人群规模评估。