Wong N, Beeso J, Sherwood R A, Peters T J
Department of Clinical Biochemistry, King's College School of Medicine and Dentistry, London, UK.
Ann Clin Biochem. 1995 Mar;32 ( Pt 2):175-80. doi: 10.1177/000456329503200207.
An extraction protocol has been developed to elute apolipoprotein B (apo B) and apolipoprotein A-1 (apo A-1) from dried blood spots with assay of the extracted apolipoproteins by automated immunoturbidimetry. Various extraction media were investigated to assess their elution efficiency and the optimum medium was found to be deionized water. Studies on the rate of elution suggested both apolipoproteins were eluted readily in less than an hour with a recovery of 71% for apo B and 65% for apo A-1. Extracted apo B but not aop A-1 was found to be stable for 24 h when kept at 4 degrees C. The within-batch coefficients of variation (CV) for the combined extraction and assay of apo B were found to be 5.9% and 8.5% at 0.6 g/L and 1.2 g/L respectively. The CVs for apo A-1 were found to be 7.4% at 0.7 g/L and 8.9% at 1.5 g/L. Antiserum from three sources (Immuno, Incstar and Bayer) was investigated for immunoreactivity with apo B and apo A-1 and cross-reactivity with red blood cells was also assessed. Although the antisera from the three companies showed similar immunoreactivity towards apo B and apo A-1, antiserum from Bayer was found to cross-react with red cells.
已开发出一种提取方案，用于从干血斑中洗脱载脂蛋白B（apo B）和载脂蛋白A-1（apo A-1），并通过自动免疫比浊法对提取的载脂蛋白进行检测。研究了各种提取介质以评估其洗脱效率，发现最佳介质是去离子水。洗脱速率研究表明，两种载脂蛋白在不到一小时内即可轻松洗脱，apo B的回收率为71%，apo A-1的回收率为65%。发现提取的apo B在4℃保存时24小时内稳定，而apo A-1则不稳定。apo B提取和检测联合操作的批内变异系数（CV）在0.6 g/L和1.2 g/L时分别为5.9%和8.5%。apo A-1在0.7 g/L时的CV为7.4%，在1.5 g/L时为8.9%。研究了来自三个来源（Immuno、Incstar和拜耳）的抗血清与apo B和apo A-1的免疫反应性，并评估了与红细胞的交叉反应性。尽管这三家公司的抗血清对apo B和apo A-1表现出相似的免疫反应性，但发现拜耳公司的抗血清与红细胞发生交叉反应。
