Yamauchi Y, Tachiband Y, Maeda A, Wakiguchi H, Usui M, Kurata T, Sairenji T
Department of Ophthalmology, Tokyo Medical College Hospital, Tokyo, Japan.
Intervirology. 1998;41(6):278-84. doi: 10.1159/000024950.
For the serodiagnosis of Epstein-Barr virus (EBV) infections, we have developed a new enzyme-linked immunosorbent assay (ELISA) for antibodies to the ZEBRA product of EBV immediate early gene BZLF1. ZEBRA protein fused with glutathione-S-transferase (GST) was expressed in Escherichia coli and purified by affinity chromatography with glutathione-Sepharose 4B. An ELISA sandwich capture system was constructed with the GST-ZEBRA immobilized on plastic microtiter plates which had been coated with a mouse monoclonal antibody to GST. ZEBRA-IgG antibodies in patients' sera with chronic active EBV infection (CAEBV) and infectious mononucleosis (IM) had, respectively, very high and high titers. Anti-ZEBRA antibodies were also detected at low titers in sera of some healthy controls. ZEBRA-IgM antibodies were detected in sera of patients with IM and CAEBV but not in sera of healthy controls. In sera of patients with CAEBV, the titers of IgG antibodies to ZEBRA correlated with the antibody titers to early antigens obtained with an immunofluorescence assay, but not to EBV nuclear antigens. This ELISA is a useful diagnostic and prognostic test for EBV infection.
为进行爱泼斯坦-巴尔病毒（EBV）感染的血清学诊断，我们开发了一种新的酶联免疫吸附测定（ELISA），用于检测针对EBV即刻早期基因BZLF1的ZEBRA产物的抗体。与谷胱甘肽-S-转移酶（GST）融合的ZEBRA蛋白在大肠杆菌中表达，并通过用谷胱甘肽-琼脂糖4B进行亲和层析纯化。构建了一种ELISA夹心捕获系统，将GST-ZEBRA固定在已用抗GST小鼠单克隆抗体包被的塑料微量滴定板上。慢性活动性EBV感染（CAEBV）和传染性单核细胞增多症（IM）患者血清中的ZEBRA-IgG抗体分别具有非常高和高的滴度。在一些健康对照者的血清中也检测到低滴度的抗ZEBRA抗体。在IM和CAEBV患者的血清中检测到ZEBRA-IgM抗体，但在健康对照者的血清中未检测到。在CAEBV患者的血清中，针对ZEBRA的IgG抗体滴度与通过免疫荧光测定获得的针对早期抗原的抗体滴度相关，但与EBV核抗原无关。这种ELISA对于EBV感染是一种有用的诊断和预后检测方法。
