SARS冠状病毒N蛋白6个不同原核表达区段的抗原特性分析  

Antigenicity characterization of six different fragments of SARS-CoV N protein expressed in E. Coli

作　　者：王慧娟[1] 周为民[1] 张陵林[1] 阮力[1] 夏宁邵[2] 谭文杰[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京100052 [2]厦门大学国家传染病诊断试剂与疫苗工程技术研究中心

出　　处：《中华实验和临床病毒学杂志》2012年第1期40-42,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：国家863课题（2007AA022464）,传染病重大专项（2008ZX10004-014）

摘　　要：目的确定原核表达的SARS—COYN蛋白不同片段抗原特性及在血清学诊断中的应用。方法在前期N蛋白抗原性初步分析的基础上，从39个原核表达不同区段的N蛋白中选取6个纯化的N蛋白即PN360（1—360aa），PN301（1—301aa），PN199（30—228aa），PN185（30—214aa），PN155b（60—214aa），PN125（90—214aa），采用Western—Boh和ELISA方法，检测SARS—COV阴性正常人血清与SARS—COV患者恢复后血清中抗SARS—COVN蛋白抗体。结果Western—Bolt结果表明6个片段皆能与SARS—COV阳性血清反应，但PN360和PN301与SARS—COV阴性血清存在明显交叉反应；使用PN199，PN185，PN155b，PN125作为包被抗原进行ELISA检测，分析表明PN185，PN155b区段的敏感性较好。结论原核表达的SARS—COVN蛋白PN185与PN155b区段是SARS—COV病毒感染的特异性抗体检测的较佳抗原。Objective To determine the antigen characteristics of different fragments of SARS-CoV N protein expressed in E. Coli and their application in the serological diagnosis. Methods Based on preliminary analysis of 39 different segments of the N protein, We choosed six purified N protein for further antigenicity characterization in this study, including that PN360 （ I - 360aa） , PN301 （ 1 - 301aa）, PN199 （30-228aa）, PN185 （30-214aa）, PN155b （60 -214aa）, and PN125 （90 -214aa）. We developed Western-Boh and ELISA to detect antibody reactivity between truncated N fragments with sera from SARS- CoV-negative normal adults or SARS-CoV patient convalescent sera. Results Western-Bolt results show that all the six fragments have reacted with the SARS patient convalescent sera, but the PN360 and PN301 showed obvious cross-reaction with sera from SARS-CoV-negative normal adults; sensitivity analysis using an ELISA coating with PN199, PN185, PN155b, PN125 as antigen showed that the PN185 and PN155b are better than PN125. Conclusion Truncated N protein PN185 and PN155b expressed in E. Coli are better antigen candidates used for detection of SARS-CoV specific antibody.

关 键 词：冠状病毒属 蛋白质N 抗原 

分 类 号：R44[医药卫生—诊断学]