SARS冠状病毒核衣壳蛋白基因的表达及其在SARS诊断中的应用

目的为SARS感染后建立一种特异性免疫学诊断方法,为基因疫苗的研制提供备选实验材料.方法利用基因重组的方法,在大肠杆菌中表达了SARS冠状病毒(SARS-CoV)N蛋白全长基因,经包涵体的初步纯化,金属螯合层析进一步纯化N蛋白,SDS-PAGE电泳和ELISA方法进行结果检测.结果N蛋白抗原与30名SARS感染患者血清结合全部为阳性,对照组30名正常人血清为阴性,30名发热但非SARS患者血清为阴性.结论利用基因重组的方法对SARS-CoV N蛋白进行了表达和纯化,证明该重组N蛋白抗原具有良好的反应特异性,是良好的应用于病毒感染早期诊断的抗原.完全可用于组构检测核心抗体的诊断试剂.并建立了免疫学的检测方法,为抗体检测提供了安全、方便的手段,并为基因工程疫苗研究奠定了基础.

Expression of SARS Coronavirus Nucleocapsid Protein Gene Fragment and Its Application of SARS Diagnosis

ObjectiveTo establish a special immune method of diagnosing SARS. To supply an alternative material of studying gene vaccine.MethodsSARS-CoV nucleocapsid protein full-length gene was expressed in E.coli by gene engineering. The fusion protein was purified by NAT chromatography, and identified by SDS-PAGE and ELISA.ResultsN protein antigen reacted with serum from 30 patients with SARS, all were positive. The serum from 30 healthy subjects and 30 feverish patients without SARS in control group were negative.ConclusionExpressing and purifying SARS-CoV N protein by gene engineering prove that the fusion protein has stronger antigenicity and it is a better antigen for the early-stage diagnosis of SARS. It can be used as diagnostic material for testing nuclear antibodies, so as to develop an immune test reference, to supply a safe and simple assay to detect antibodies and to make the basis of studying gene vaccine.