CAJ | 학술논문

目的确定原核表达的SARS-COV N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-Bolt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-COV N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-COV N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.
목적 학정원핵표체적SARS-COV N단백불동편단항원특성급재혈청학진단중적응용.방법 재전기N단백항원성초보분석적기출상,종39개원핵표체불동구단적N단백중선취6개순화적N단백즉PN360(1 -360aa),PN301(1- 301aa),PN199( 30 - 228aa),PN185(30 - 214aa),PN155b(60 - 214aa),PN125 (90 - 214aa),채용Western-Bolt화ELISA방법,검측SARS-COV음성정상인혈청여SARS-COV환자회복후혈청중항SARS-COV N단백항체.결과 Western-Bolt결과표명6개편단개능여SARS-COV양성혈청반응,단PN360화PN301여SARS-COV음성혈청존재명현교차반응;사용PN199,PN185,PN155b,PN125작위포피항원진행ELISA검측,분석표명PN185,PN155b구단적민감성교호.결론 원핵표체적SARS-COV N단백PN185여PN155b구단시SARS-COV병독감염적특이성항체검측적교가항원.
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 (1 -360aa),PN301 (1 -301aa),PN199 (30-228aa),PN185 (30-214aa),PN155b (60-214aa),and PN125 (90-214aa).We developed Western-Bolt and ELISA to detect antibody reactivity between truncated N fragments with sera from SARSCoV-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.