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目的 考察具有多个优势抗原表位区段嵌合表达的戊型肝炎病毒（HEV）重组抗原的免疫学性质。方法 将构建的含有3个不同HEV抗原表位基因(ORF2．1: 6287 ～ 6403nt, ORF2．2: 6743 ～ 7126nt, ORF3)的表达质粒pThioHisORF(2．1＋2．2＋3 )在大肠杆菌中进行异丙基硫代半乳糖苷（IPTG）诱导表达，其表达产物P_(2．1＋2．2＋3 )在8 mol/L尿素下经阳离子柱SP Sepharose FF层析纯化，透析复性。以P_(2．1＋2．2＋3 )免疫家兔和小鼠, 通过ELISA检测实验动物血清及戊型肝炎患者阳性血清IgG抗体，以Western蛋白印迹检测动物免疫血清对杆状病毒系统表达的抗原片段及嵌合抗原P_(2．1＋2．2＋3 )对患者阳性血清的免疫反应性。结果 纯化的抗原纯度达90%以上；免疫动物血清中检测到特异性抗体的滴度分别为1∶25 600（家兔）和1∶12 800（小鼠）以上，抗血清能够特异识别杆状病毒系统表达的HEV抗原片段，P_(2．1＋2．2＋3 )能与患者阳性血清特异反应。结论 嵌合表达的HEV重组抗原P_(2．1＋2．2＋3 )具有良好的免疫原性和免疫反应性，为进一步研究开发HEV诊断试剂盒及基因工程疫苗提供了基础。
Objective To investigate the immunological properties of hepatitis E virus (HEV) recombinant antigens with chimeric expression of multiple dominant epitopes. Methods The constructed expression plasmid pThioHisORF (2.1 + 2.2 + 3) containing 3 different HEV epitope genes (ORF2.1: 6287 ~ 6403nt, ORF2.2: 6743 ~ 7126nt, ORF3) The expression of P_ (2.1 + 2.2 + 3) was induced by IPTG and purified by cation exchange chromatography (SP Sepharose FF) under 8 mol / L urea. The rabbits and mice were immunized with P (2.1 + 2.2 + 3), and the positive serum IgG antibodies in serum of experimental animals and patients with hepatitis E were detected by ELISA. The Western blot was used to detect the expression of antigenic fragments of the animal immune system against baculovirus Immunoreactivity of chimeric antigen P_ (2.1 + 2.2 + 3) to patient positive serum. Results The purity of the purified antigen was above 90%. The titer of the specific antibody in the serum of immunized animals was 1:25 600 (rabbit) and 1:12 800 (mouse), respectively. The antiserum could specifically recognize the rod The virus system expressed HEV antigen fragment, P_ (2.1 + 2.2 +3) can react specifically with the patient’s positive serum. Conclusion The chimeric recombinant HEV recombinant P_ (2.1 + 2.2 + 3) has good immunogenicity and immunoreactivity, which provides a basis for further research and development of HEV diagnostic kits and genetic engineering vaccines.