Kurosaki Yoshifumi, Hachimura Kazuo, Ogawa Zensuke
Department of Molecular Medical Biology, the Graduate School of Medical Sciences, Kitasato University, Kanagawa, Japan.
Clin Chim Acta. 2009 Mar;401(1-2):110-3. doi: 10.1016/j.cca.2008.11.026. Epub 2008 Dec 6.
2 homogeneous assay kits, Determiner L HDL-C from Kyowa Medex, Co., Ltd. (Tokyo, Japan) (KWM) and Cholestest NHDL from Daiichi Pure Chemical, Co., Ltd. (Tokyo, Japan) (DIC), for HDL-C are widely used in laboratory tests worldwide. Meanwhile, it was reported that free cholesterol (FC) in HDL is not measured with one of these kits. We devised a novel evaluation method and analyzed the 2 kits in detail.
Using HPLC, the residual amounts of cholesterol in reaction mixtures were compared before and after reaction with homogeneous reagents. Healthy sera were reacted with homogeneous reagents or with heated, non-enzyme reagents from each kit. The FC and total cholesterol in each lipoprotein were continuously measured with HPLC.
With KWM, the HDL-C was measured >95%.With DIC, all FC in sera was eliminated in the first reaction, and FC in HDL was not measured. Moreover, the peak X that was assumed to be a part of HDL was separated with DIC, and the cholesterol in the peak X was not measured.
2 HDL-C assay kits were compared using a novel evaluation method. KWM was good overall. DIC was found to have 2 problems: 1) FC in HDL was not measured, and 2) the cholesterol in peak X that was assumed to be a part of HDL was not measured.
两种均相检测试剂盒，即日本东京协和梅迪克公司（Kyowa Medex, Co., Ltd.）的Determiner L HDL-C（KWM）和日本东京第一制药纯化学公司（Daiichi Pure Chemical, Co., Ltd.）的Cholestest NHDL（DIC），用于检测高密度脂蛋白胆固醇（HDL-C），在全球实验室检测中广泛使用。同时，有报道称其中一种试剂盒无法检测HDL中的游离胆固醇（FC）。我们设计了一种新的评估方法，并对这两种试剂盒进行了详细分析。
使用高效液相色谱法（HPLC），比较反应混合物与均相试剂反应前后胆固醇的残留量。将健康血清与均相试剂或与来自每个试剂盒的加热的非酶试剂反应。用HPLC连续测量每种脂蛋白中的FC和总胆固醇。
使用KWM时，HDL-C的检测率>95%。使用DIC时，血清中的所有FC在第一次反应中被消除，HDL中的FC未被检测到。此外，被认为是HDL一部分的峰X被DIC分离，峰X中的胆固醇未被检测到。
使用一种新的评估方法对两种HDL-C检测试剂盒进行了比较。总体而言，KWM表现良好。发现DIC有两个问题：1）未检测到HDL中的FC；2）未检测到被认为是HDL一部分的峰X中的胆固醇。
