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以痘苗病毒为载体克隆了 Epstein-Barr 病毒(EBV)壳抗原(VCA)的基因,重组痘苗病毒在感染细胞表达蛋白产物。用间接免疫荧光、免疫印迹技术和免疫电镜确定反应的特异性,表达产物位于细胞浆内和核内,胞浆内大量集聚,致使核偏向一侧,分子量约为100000。重组痘苗病毒免疫血清能特异性地与 B95-8细胞浆内的抗原反应。用重组病毒感染的细胞为靶细胞,并以 B95-8细胞和天坛株痘苗病毒感染的细胞分别为阳性和阴性对照,用免疫荧光方法平行检查不同人群血清中的VCA/IgG 和 VCA/IgA 抗体。结果表明,基因工程表达产物的特异性和在检查人血清抗体中的意义,为 ELISA 的建立和应用提供可靠的依据和抗原来源。
The gene of Epstein-Barr virus (EBV) shell antigen (VCA) was cloned using vaccinia virus and the recombinant vaccinia virus expressed the protein product in infected cells. Indirect immunofluorescence, immunoblotting and immunoelectron microscopy were used to determine the specificity of the reaction. The expressed product was localized in the cytoplasm and nucleus and accumulated abundantly in the cytoplasm, causing the nucleus to deviate to one side with a molecular weight of about 100,000. Recombinant vaccinia virus serum can specifically react with antigens in B95-8 cytoplasm. Cells infected with the recombinant virus were used as target cells. Cells infected with vaccinia virus of B95-8 cells and Tiantan strain were positive and negative controls, and VCA / IgG and VCA / IgA antibodies in serum of different populations were examined in parallel by immunofluorescence . The results showed that the specificity of the gene expression product and the significance in the examination of human serum antibody provided a reliable basis and antigen source for the establishment and application of ELISA.