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作 者:谷淑燕[1] 王轶龙 李燕[1] 朱秋屏[1] H.Wolf
机构地区:[1]中国预防医学科学院病毒学研究所,北京100052 [2]德国慕尼黑大学
出 处:《中华实验和临床病毒学杂志》1996年第1期62-66,共5页Chinese Journal of Experimental and Clinical Virology
摘 要: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 major component of Epstein-Barr viral capsid antigens(EBV-VCA)was expressed in mammalia cells and used for detection of antibody in human sera.The full ORF was constructed and transfected in- to vaccinia viruses.The expressing antigen on cells infected with recombinant vaccinia viruses was characterized by immunofluorecence test,Western-blot and immuno-electronmicroscope.The specific antigen localized in cytoplasm and nucleus and mainly in cytoplasm.The serum from vaccinated rabbit with recombinant vaccinia viruses specifically reacted with the antigen in B95-8 cells.Cells infected with recombinant vaccinia virus were used for de- tection of antibody to EBV-VCA and compared with B95-8 cells.The cell expressed antigen are useful and more sensitive for detectiou of VCA/IgG antibody and screening VCA/IgA antibody positive persons from NPC high risk area.
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