三种人类8型疱疹病毒特异性抗原表达及复合抗原血清学检测方法的建立  

Establishment of serological detection methods of 3 types of human herpesvirus 8 specific-antigens and complex antigens

作　　者：王星[1] 何方平[1] 卢晓梅[1] 赵淑君[1] 林仁勇[1] 何斌 温浩[1] 

机构地区：[1]新疆医科大学第一附属医院中心实验室 [2]Department of Microbiology and Immunology College of Medicine University of Illinois,Chicago

出　　处：《中华传染病杂志》2007年第9期562-566,共5页Chinese Journal of Infectious Diseases

基　　金：新疆维吾尔自治区自然科学基金(200421122);新疆维吾尔自治区高校科研计划创新群体资助项目(XJEDU2004G10)

摘　　要：目的建立人类8型疱疹病毒(HHV-8)血清学检测办法。方法利用E.coli系统合成HHV-8三个抗原性较强的融合蛋白:K8.1,ORF65和ORF73 C,作为检测抗原,阳性血清为本地12份卡波西肉瘤患者血清和32份AIDS相关型卡波西肉瘤患者血清;阴性血清为20份皮肤肿瘤患者血清和77份15岁以下儿童血清。以Western印迹及ELISA法确定各重组蛋白免疫原性和该混合抗原ELISA法的敏感度和特异度。结果获得高纯度的3种8型疱疹病毒特异重组蛋白,均具备较强的抗原特性。应用上述复合抗原ELISA法与传统免疫荧光法共同筛查同批次阳性和阴性血清,发现前者敏感度远高于后者,分别为81.8%和34.4%,特异度则为97.9%。结论K8.1、ORF65、ORF73 C是建立HHV-8血清学检测方法较佳的候选抗原,有更高的敏感度和特异度。Objective To establish serological detection methods of human herpesvirus 8 （HHV-8）. Methods Three potent antigenic fusion proteins, K8.1, ORF65 and ORF73 C of HHV- 8 were synthesized using E. coli system. The sera were detected using these antigenic proteins. The positive sera were from 12 patients with Kaposi＇s sarcoma and 32 patients with acquired immunodeficiency syndrome-related Kaposi＇s sarcoma. The negative sera were from 20 patients with cutaneous tumors and children under 15 years old. Western-blot and enzyme-linked imrnunosorbent assay （ELISA） were employed to determine the immunogenicity of each recombinant protein and the sensitivity and specificity of ELISA using the complex antigens. Results Three types highly purified HHV-8 specific recombinant pro- teins with potent antigenicity were successfully synthesized. The sensitivity of ELISA using the above complex antigens was significantly higher than traditional immuno-flurescent assay （IFA） detecting the positive and negative sera, which were 81.8 %, 34.4 %, respectively. And the specificity of ELISA was 97.9%. Condusion K8. 1, ORF65 and ORF73 C are good candidate antigens for establishing HHV-8 serological detection methods, which have better sensitivity and specificity.

关 键 词：肉瘤 卡波西 疱疹病毒8型 人 基因表达 抗原 酶联免疫吸附测定 重组融合蛋白质类 

分 类 号：R446.6[医药卫生—诊断学]