摘要
本研究构建了SARS冠状病毒纤突蛋白(S)重组杆状病毒(rBac-SS)。SDS-PAGE及Western-Blot分析表明约190 Ku左右的重组SARS纤突蛋白(rSS)在rBac-SS感染sf9昆虫细胞获得表达,并具有特异免疫反应原性。以rBac-SS感染的昆虫细胞裂解物稀释后直接包被ELISA板,与Vero细胞培养的全病毒裂解物比较,检测SARS-CoV康复病人血清特异抗体,表现出同样的敏感性和特异性;rBac-SS感染的昆虫细胞用于间接免疫荧光,快速检测血清特异抗体反应,具有良好的敏感性和特异性。结果显示,杆状病毒表达的rSS有望替代SARS-CoV全病毒,作为安全、敏感和特异的重组诊断抗原,并为探索重组亚单位疫苗的可行性奠定基础。
A recombinant baculovims（ rBac-SS）expressing spike protein of severe acute respiratory syndrome coronavirus was constmcted. The expression of full length recombinant spike protein （rSS） in insect cells was confirmed by SDS-PAGE and Western-blot （ 190 Ku）. When lysis supematant of rBac-SS infected insect cells was directly used in ELISA as coating antigen for detecting specific antibodies in convalesce serum of SARS-CoV infected patient, the rSS showed the same sensitivity as the cell lysis of veto E6 infected with SARS-CoV, Insect cell infected with rBac-SS also showed highly sensitivity to detect specific serum antibody by indirect inmmnofluoresence assay （IFA）.The results here indicated that baculovims expressed rSS is promising to replace the whole SARS-CoV as antigen in ELISA and IFA for detecting the SARS-CoV specific antibodies and potential subtmit vaccine candidate.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2005年第6期549-552,共4页
Chinese Journal of Preventive Veterinary Medicine
