Siedel J, Schiefer S, Rosseneu M, Bergeaud R, De Keersgieter W, Pautz B, Vinaimont N, Ziegenhorn J
Boehringer Mannheim GmbH, Biochemical Research Center, Tutzing, F.R.G.
Clin Chem. 1988 Sep;34(9):1821-5.
We describe a method for routine immunoturbidimetry of apolipoproteins (apo) A-I, A-II, and B in both normo- and hyperlipemic sera. A special antiserum reagent, consisting of a highly concentrated mixture of nonionic and anionic detergents (final concentration in the assay, 36 g/L), rapidly removes intrinsic turbidities of even strongly lipemic sera without interfering with the antigen-antibody precipitation reaction. The method has good precision, and obviates the need for special sample pretreatment, extended incubation periods, and measurment of sample blanks. A comparison with established immunoephelometric assays generally showed close agreement for analytical recoveries of the three apolipoproteins. However, in samples containing greater than or equal to 18 g of triglycerides per liter, the nephelometric assays yielded about two- to threefold higher values for apo A-II and B than did the turbidimetric procedure. To elucidate this discrepancy, we used the turbidimetric methods to assay sera with and without enzymatic lipolytic pretreatment. Even for samples with triglyceride concentrations up to 60 g/L, complete enzymatic lipolysis (as evidenced by thin-layer chromatography) did not significantly alter the recoveries of apo A-II and B from those obtained with the untreated specimens. Thus the immunoturbidimetric methods yield reliable results for apo A-I, A-II, and B, not only in normo- but also in hyperlipemic sera.
我们描述了一种用于常规免疫比浊法检测正常血脂和高血脂血清中载脂蛋白(apo)A-I、A-II和B的方法。一种特殊的抗血清试剂,由高浓度的非离子和阴离子洗涤剂混合物组成(测定中的终浓度为36 g/L),能快速消除即使是高度脂血血清的固有浊度,而不干扰抗原-抗体沉淀反应。该方法具有良好的精密度,无需特殊的样品预处理、延长孵育时间和测定样品空白。与已建立的免疫比浊法比较,通常显示三种载脂蛋白的分析回收率密切一致。然而,在每升含甘油三酯大于或等于18 g的样品中,比浊法测定的apo A-II和B值比浊度法高出约两到三倍。为了阐明这种差异,我们用比浊法检测了经酶促脂解预处理和未预处理的血清。即使对于甘油三酯浓度高达60 g/L的样品,完全酶促脂解(通过薄层色谱证明)也不会显著改变apo A-II和B与未处理标本相比的回收率。因此,免疫比浊法不仅对正常血脂血清,而且对高血脂血清中的apo A-I、A-II和B都能产生可靠的结果。