Ohta T, Migita M, Yasutake T, Matsuda I
Department of Pediatrics, Kumamoto University Medical School, Japan.
J Pediatr Gastroenterol Nutr. 1988 Jul-Aug;7(4):524-31. doi: 10.1097/00005176-198807000-00008.
Hypercholesterolemia is a risk factor for atherosclerotic cardiovascular disease, and early diagnosis and treatment should be given attention. We developed a simple and sensitive enzyme-linked immunosorbent assay (ELISA) for apolipoprotein B (apoB) on a dried blood spot (DBS). The specificity of this assay was investigated by constructing curves for other proteins. The cross-reactivity was negligible. The mean intra- and intercoefficients of variation were 5.2 and 7.8%, respectively. We used phosphate buffer with Triton X-100 for extracting apoB on DBS. The elution of apoB was stable up to 30 days after bleeding. The correlation between plasma and DBS apoB was r = 0.94, p less than 0.005, n = 55. Using this method, we screened 2,500 babies between 5 and 7 days of age. The mean +/- SD of DBS apoB was 75 +/- 15 U. Seven hypercholesterolemic neonates were detected. Family studies of these neonates disclosed one definite and two suspected heterozygotes for familial hypercholesterolemia. Four other neonates showed no related familial background. This ELISA method for assaying apoB should prove to be a useful tool for large mass screening for hypercholesterolemia, particularly in the newborn.
高胆固醇血症是动脉粥样硬化性心血管疾病的一个危险因素，应重视早期诊断和治疗。我们开发了一种用于检测干血斑（DBS）中载脂蛋白B（apoB）的简单且灵敏的酶联免疫吸附测定（ELISA）方法。通过构建其他蛋白质的曲线来研究该测定方法的特异性。交叉反应可忽略不计。平均批内和批间变异系数分别为5.2%和7.8%。我们使用含有 Triton X - 100的磷酸盐缓冲液从DBS中提取apoB。出血后30天内apoB的洗脱是稳定的。血浆和DBS中apoB的相关性为r = 0.94，p < 0.005，n = 55。使用该方法，我们对2500名5至7日龄的婴儿进行了筛查。DBS中apoB的平均值±标准差为75±15 U。检测到7名高胆固醇血症新生儿。对这些新生儿的家族研究发现1名家族性高胆固醇血症的明确杂合子和2名疑似杂合子。另外4名新生儿未显示相关家族背景。这种检测apoB的ELISA方法应被证明是大规模筛查高胆固醇血症的有用工具，尤其是在新生儿中。
