Ellins M L, Campbell J B
J Clin Microbiol. 1977 Oct;6(4):348-58. doi: 10.1128/jcm.6.4.348-358.1977.
By using trypsin-treated human type O cells as indicators, we compared the abilities of four polyanion-divalent cation combinations (heparin-MnCl(2); high-and low-molecular-weight dextran sulfate-CaCl(2); and sodium polyanetholesulfonate [SPS]-CaCl(2)) for removal of serum non-immunoglobulin (lipoprotein) inhibitors of rubella hemagglutination. The combination of SPS-CaCl(2) was found to be the most effective, precipitating completely the pre-beta and beta-lipoproteins and reducing the alpha-lipoprotein levels by more than 50%. Hemagglutination patterns after this treatment were clear and stable, and, when normal sera were tested, hemagglutination-inhibition (HI) titers were comparable to those obtained after standard heparin-MnCl(2) treatment. High-molecular-weight dextran sulfate-CaCl(2) removed serum lipoproteins almost as effectively as SPS-CaCl(2). However, problems of nonspecific agglutination and the heavy hemagglutination patterns resulting made this combination unacceptable for routine purposes. Neither low-molecular-weight dextran sulfate-CaCl(2) nor heparin-MnCl(2) removed the pre-beta lipoproteins completely, and occasionally traces of beta-lipoprotein also remained after treatment. The presence of pre-beta lipoproteins in normal sera after treatment may be of no consequence in the HI test since we have found that the very-low-density lipoprotein fractions obtained by ultracentrifugal methods from normal sera (those corresponding to the pre-beta fractions obtained by electrophoresis) had no HI activity. However, very-low-density lipoprotein fractions from all hyperlipemic sera tested had HI activity (titers ranging from 1:16 to 1:1,024) which, in the majority of cases, was not eliminated after heparin-MnCl(2) treatment. In every case, treatment with SPS-CaCl(2) removed this nonspecific activity completely. Since hyperlipemic sera may occasionally be encountered in routine rubella HI antibody testing, we recommend the use of SPS-CaCl(2) rather than heparin-MnCl(2) for pretreatment of sera.
以经胰蛋白酶处理的人O型细胞为指示物，我们比较了四种聚阴离子 - 二价阳离子组合（肝素 - 氯化锰；高、低分子量硫酸葡聚糖 - 氯化钙；以及多聚茴香脑磺酸钠[SPS] - 氯化钙）去除血清中风疹血凝反应非免疫球蛋白（脂蛋白）抑制剂的能力。结果发现，SPS - 氯化钙组合最为有效，能使前β脂蛋白和β脂蛋白完全沉淀，并使α脂蛋白水平降低50%以上。该处理后的血凝模式清晰且稳定，检测正常血清时，血凝抑制（HI）效价与标准肝素 - 氯化锰处理后获得的效价相当。高分子量硫酸葡聚糖 - 氯化钙去除血清脂蛋白的效果几乎与SPS - 氯化钙相同。然而，非特异性凝集问题以及由此导致的严重血凝模式使其不适用于常规用途。低分子量硫酸葡聚糖 - 氯化钙和肝素 - 氯化锰均不能完全去除前β脂蛋白，处理后偶尔还会残留微量的β脂蛋白。处理后正常血清中前β脂蛋白的存在在HI试验中可能无关紧要，因为我们发现通过超速离心法从正常血清中获得的极低密度脂蛋白组分（对应于通过电泳获得的前β组分）没有HI活性。然而，所有检测的高脂血症血清中的极低密度脂蛋白组分都有HI活性（效价范围为1:16至1:1024），在大多数情况下，肝素 - 氯化锰处理后这种活性并未消除。在每种情况下，SPS - 氯化钙处理都能完全去除这种非特异性活性。由于在常规风疹HI抗体检测中偶尔会遇到高脂血症血清，我们建议使用SPS - 氯化钙而非肝素 - 氯化锰对血清进行预处理。
