Lee F K, Nahmias A J, Nahmias D G, McDougal J S
J Virol Methods. 1983 Sep;7(3):167-81. doi: 10.1016/0166-0934(83)90006-x.
A method has been developed to detect virus particles within immune complexes by electron microscopy (VICEM test). Radioiodinated or unlabeled adenovirus type 2 immune complexes (IC) in human serum were used to determine the optimal conditions. Best results were obtained when the immune complexes were precipitated with 5% polyethylene glycol 6000, adsorbed with 0.5% of protein A-containing Staphylococcus aureus (Cowan 1) and eluted, with brief sonication, in 2 M propionic acid. When the eluate was examined by the pseudoreplica method of electron microscopy, morphologically intact viruses, concentrated over five-fold and free from background serum proteins, were readily demonstrated. The method is applicable to virus IC with a wide-range of antigen-to-antibody ratios and requires a minimum of 10(4)--10(5) virus particles. This method should be useful for the characterization of IC in clinical specimens obtained from patients with viral infections and for the possible demonstration of viral agents in immune complex diseases of unknown etiology. A modification of the technique can also be used for the rapid serological identification of viruses by electron microscopy, as exemplified by studies using adeno- and herpes simplex viruses.
已开发出一种通过电子显微镜检测免疫复合物中病毒颗粒的方法(VICEM检测)。使用人血清中放射性碘化或未标记的2型腺病毒免疫复合物(IC)来确定最佳条件。当免疫复合物用5%聚乙二醇6000沉淀、用0.5%含蛋白A的金黄色葡萄球菌(考恩1株)吸附并通过短暂超声处理在2M丙酸中洗脱时,可获得最佳结果。当通过电子显微镜的假复型法检查洗脱液时,形态完整的病毒很容易被证明,其浓度提高了五倍以上且无背景血清蛋白。该方法适用于抗原与抗体比例范围广泛的病毒IC,并且至少需要10⁴ - 10⁵个病毒颗粒。该方法对于表征从病毒感染患者获得的临床标本中的IC以及可能在病因不明的免疫复合物疾病中证明病毒病原体应该是有用的。该技术的一种改进也可用于通过电子显微镜对病毒进行快速血清学鉴定,如使用腺病毒和单纯疱疹病毒的研究所举例说明的那样。