Weiblen B J, Schumacher R T, Hoff R
Massachusetts Department of Public Health, State Laboratory Institute, Jamaica Plain 02130.
J Immunol Methods. 1990 Feb 9;126(2):199-204. doi: 10.1016/0022-1759(90)90151-k.
Indirect assays for IgM and IgA antibodies often lack sensitivity and specificity due to interference from IgG antibodies. To overcome this problem we have developed a simple procedure using recombinant protein G coupled to agarose beads to remove the interfering IgG. A series of HIV seroconversion panels was tested by IgM and IgA immunoblot after protein G treatment in order to evaluate IgG removal and to study appearance of IgM and IgA antibodies in early HIV infection. Protein G treatment removed 99.9% of the IgG and reduced IgG anti-HIV titers of over 1/100,000 to undetectable levels. Both IgM and IgA HIV antibodies were detected as early in seroconversion as were IgG HIV antibodies. IgA HIV antibodies persisted for a longer period of time, reacted with more HIV proteins, and showed more intense staining than IgM HIV antibodies.
由于受到IgG抗体的干扰,针对IgM和IgA抗体的间接检测方法常常缺乏敏感性和特异性。为克服这一问题,我们开发了一种简单的方法,即使用与琼脂糖珠偶联的重组蛋白G来去除干扰性IgG。对一系列HIV血清转化样本在经蛋白G处理后进行IgM和IgA免疫印迹检测,以评估IgG的去除情况,并研究早期HIV感染中IgM和IgA抗体的出现情况。蛋白G处理去除了99.9%的IgG,并将超过1/100,000的IgG抗HIV滴度降低至检测不到的水平。在血清转化过程中,IgM和IgA HIV抗体与IgG HIV抗体一样在早期就能被检测到。IgA HIV抗体持续的时间更长,与更多的HIV蛋白发生反应,并且比IgM HIV抗体显示出更强的染色。