Developments and clinical evaluations of easy-to-perform and rapid measurements of serum Lp(a) concentrations and apo(a) isoforms.

虽然开发了几种不同的测定血清脂蛋白(a) （Lp(a)）的技术，但许多实验室仍然缺乏可靠的、易于执行的Lp(a)测定方法。目前，ELISA法是测定血清Lp(a)最常用的技术，因为它灵敏度高，原则上具有特异性，并且不需要放射性同位素。然而，ELISA方法需要高度稀释的血清作为样品，一式两份测量以达到令人满意的精度，并且需要数小时的分析时间。免疫浊度法和免疫浊度法适用于临床实验室，因为它们快速且易于执行，具有高通量，并且可以自动化。然而，浊度免疫测定（TIA）和乳胶增强免疫测定（LIA）方法对被测分析物的大小差异也很敏感。经过仔细的评估，我们选择了用LIA方法测量Lp(a)的自动化程序，具有令人满意的精度，特异性和灵敏度。另一方面，一个新的系统已被用来确定载脂蛋白(a)表型。电泳和免疫印迹后，用计算机数字微标仪测量载脂蛋白(a)异型带的Rf值，并根据Rf值自动分类载脂蛋白(a)表型。最近，为了对载脂蛋白(a)基因型进行分类，我们利用限制性内切酶kpni和载脂蛋白(a) kringle IV特异性探针对基因组DNA进行脉冲场凝胶电泳，研究了载脂蛋白(a)大小多态性的基础。

Although several different techniques to determine serum lipoprotein(a) (Lp(a)) were developed, many laboratories still lack reliable any easy-to-perform assay for Lp(a) measurements. At present, ELISA method is the most commonly used technique to measure serum Lp(a), because it is highly sensitive, specific in principle, and does not require radio-isotopes. Gowever, the ELISA method requires highly diluted serum as samples, measurement in duplicate to achieve satisfactory precision, and several hours for the assay. The immunonephelometric and immunoturbidimetric assays are suitable for the clinical laboratories, in that they are rapid and easy-to-perform, have a high throughput, and can be automated. The turbidimetric immunoassay (TIA) and latex-enhanced immunoassay (LIA) methods, however, also are sensitive to differences in size of the analyte being measured. After the careful evaluations, we have chosen automated procedure for Lp(a) measurement by an LIA method having satisfactory precision, specificity and sensitivity.On the other hand, a new system has been employed to determine the apo(a) phenotypes. After electrophoresis and immunoblotting, Rf values of apo(a) isoform bands were measured using a computerized digital micro scale and apo(a) phenotypes were categorized automatically based on the Rf values. Recently, in an attempt to classify the apo(a) genotypes, we have investigated the basis of the apo(a) size polymorphism by pulsed-field gel electrophoresis of genomic DNA employing the restriction enzymeKpnI and an apo(a) kringle IV specific probe.