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技术来测定血清脂蛋白（a）（Lp（a）），许多实验室仍然缺乏可靠的任何易于执行的测定Lp（a）的测量。酶联免疫吸附法（ELISA）是目前最常用的血清Lp（a）测定方法，因为它灵敏度高，原则上特异性强，且不需要放射性同位素。甘精胰岛素，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密度，并且测定需要数小时。免疫比浊法和免疫比浊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，比浊免疫测定（TIA）和胶乳增强免疫测定（LIA）方法也对被测量的分析物的大小差异敏感。经过仔细的评估，我们选择了一种精密度、特异性和灵敏度均令人满意的LIA法用于Lp（a）的自动化检测，另一方面，我们采用了一种新的系统来测定载脂蛋白（a）表型。在电泳和免疫印迹后，使用计算机化的数字化微标测量apo（a）同种型条带的Rf值，并基于Rf值自动对apo（a）表型进行分类。最近，为了对载脂蛋白（a）基因型进行分类，我们采用限制性内切酶Kpn I和载脂蛋白（a）Kringle IV特异性探针，通过基因组DNA的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.