乙型肝炎病毒重组抗原表位检测敏感性研究

目的:在外源抗原表位表达载体系统中构建并表达乙型肝炎病毒(HBV)抗原表位,并对其诊断敏感性进行评价。方法:人工合成编码HBV“a”抗原决定簇表位中24个氨基酸(S124-147)的寡核苷酸序列,克隆入外源抗原表位表达载体系统FHV-RNA2-I3位点,构建重组质粒,在原核pET系统(BL21细胞)中表达。通过ELISA和West-ernblot检测表达产物的抗原性,并以表达产物为包被抗原,通过ELISA和Westernblot检测58份乙肝患者血清抗-HBs。结果:嵌和蛋白可与抗-HBs特异性结合,其ELISA检测抗-HBs阳性率为77.5%,Westernblot的为68%,而对照试剂盒的为62%。结论:在外源抗原表位表达系统中表达的HBV重组抗原表位具有良好的抗原性,有诊断应用价值。

Study on Antigenicity of HBV Recombinant Epitope

Objective Construction and Expression HBV recombinant epitope in a foreign epitope presenting vector,and evalution of dignostic sensitivity for anti-HBs with HBV epitope. Methods Oligonucleotids encoded twenty-four amino acids(aaS124-147) which is within "a" antigenic deterninant were synthesized and inserted into a foreign epitope-presenting carrier I3 site (FHV-RNA2-cDNA-I3). HBV epitope was expressed in a prokaryotic cellular system(BL21 cell). The expression product was analyzed and its antigenicity was further studied by ELISA and Wstern blot method with chimeric protein as coating antigen.we used chimeric protein as coating antigen to test anti-HBs with ELISA and Wstern blot in 58 seral samples. Results The chimeric protein reacted with anti-HBs seral specially.The detection positive ratio of anti-HBs is 77.5% and 68%respectively by ELISA and Wstern blot with chimeric protein as coating antigen,the control's was 62%.Conclusion HBV epitope expressed in a foreign epitope-presenting carrier possessed good antigenicity and diagnostic value.