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目的获得重组SARS冠状病毒(SARS-CoV)N蛋白抗原,建立特异性诊断SARS病毒感染的免疫学方法。方法大肠埃希菌中表达SARS病毒N蛋白基因,用金属螯合层析纯化N蛋白,建立检测SARS抗体的ELISA方法。结果大肠埃希菌中表达了SARS病毒全长N蛋白抗原,经包涵体洗涤和金属螯和纯化后得到纯度较高的重组蛋白。用重组抗原ELISA检测30名SARS患者抗体全部为阳性,30名正常人血清为阴性,30名发热非SARS患者血清为阴性。结论SARS表达核蛋白可以在大肠埃希菌中得到高效表达,纯化的重组N蛋白具有良好抗原性,可用于检测SARS抗体。
Objective To obtain recombinant SARS-CoV (N-protein) antigens of SARS-CoV and establish an immunological method to diagnose the SARS-specific virus infection. Methods The SARS virus N protein gene was expressed in Escherichia coli and the N protein was purified by metal chelation chromatography. The ELISA method for detecting SARS virus was established. Results The full-length N protein antigen of SARS virus was expressed in Escherichia coli, and the recombinant protein with high purity was obtained after washing with inclusion body and metal chelation and purification. Antibodies against all 30 SARS patients were all positive by recombinant antigen ELISA. Serum samples from 30 normal people were negative. Thirty patients with non-SARS fever were negative. Conclusion SARS-expressing nucleoprotein can be efficiently expressed in Escherichia coli. The purified recombinant N protein has good antigenicity and can be used to detect SARS antibodies.