人冠状病毒HKU1血清学检测方法的建立及其初步应用  

Development of serological detection assays for human coronavirus HKU1 infection and its application

作　　者：周为民[1] 王文玲[1] 谭文杰[1] 张陵林[1] 殷霄[1] 陆柔剑[1] 王慧娟[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

出　　处：《中华实验和临床病毒学杂志》2010年第5期376-379,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：863课题（2007AA02Z464,2007AA02Z463）;十一五重大专项（2008ZX10004-014）

摘　　要：目的 表达HCoV-HKU1的壳蛋白（N蛋白）及棘突蛋白（S蛋白）,建立检测血清中相应抗体的方法.方法 使用原核表达系统表达纯化HCoV-HKU1的N蛋白,转膜制条建立基于蛋白印迹的N抗体检测方法.构建HCoV-HKU1 S蛋白的真核表达质粒,转染细胞后,用间接免疫荧光（IFA）建立S抗体检测方法.利用已建立的N抗体和S抗体检测方法,检测了100份正常成人血清.结果 经Western Blot检测,S和N蛋白表达正确;利用已建立的血清学检测方法分析100份正常成人血清,其中HCoV-HKU1 S抗体的阳性率为47%,N抗体的阳性率为48哂,S抗体和N抗体均阳性的占总数的21%,双抗体均阴性的占总数的22%.S抗体和N抗体共同检测可获得74%的阳性检出率.结论 所建立的方法可用于HCoV-HKU1血清学分析,共同检测HCoV-HKU1 S抗体和N抗体可获得较好的检测结果.在正常成年中HCoV-HKU1抗体阳性检出率较高.Objective To express the nuclear capsid protein （N protein） and the spike protein （S protein） of HCoV-HKU1, and to develop the corresponding serum assay for antibody detection. Methods The N protein of HCoV-HKU1 was expressed in E. Coli, anti-N antibody assay was established using Western Blotting with turn-based membrane. HCoV-HKU1 S protein was constructed in the eukaryotic expression plasmids, and confirmed by Western Blotting, S antibody assay was established using indirect immunofluorescence assay （IFA）. We analyzed anti-S and anti-N antibody among 100 normal adult serum.Results Expression of S and N protein were confirmed;100 normal adult serum were analyzed using the established serological detection assay, in which HCoV-HKU1 S antibody positive rate was 47% , N antibody positive rate was 48% , Both S and N antibodies positive were 21% , Both S and N antibodies negative were 22%. Co-detection S and N antibody was achieved 74% positive rate. Conclusion The methods we established here could be used for serological analysis of HCoV-HKU1. Either detection of HCoV-HKU1 S or N antibodies achieved good results. Higher positive detection rate of anti-S or anti-N antibody was found in the normal adults.

关 键 词：冠状病毒属 病毒蛋白质类 抗体 病毒 血清学 

分 类 号：R446.6[医药卫生—诊断学]