摘要
目的 对戊型肝炎病毒 (HEV)Ⅳ型ORF3的全长基因片段和 4个覆盖全长ORF2的互相重叠的基因片段进行了表达 ,对表达产物进行了纯化及抗原性分析。方法 将O3、FB5、E4、F2 2和E5基因片段分别连接到融合性表达质粒pThioHis,用IPTG进行诱导表达 ,并用反向、分子筛、离子交换和亲和层析方法进行纯化 ,用纯化的蛋白分别制备检测抗HEVIgG的诊断试剂 ,用该试剂检测急性散发性的戊肝病人和非戊肝病人血清。结果 位于ORF2N 端的FB5蛋白在急性期的戊肝病人血清中具有最强的反应性 ;而且仍有部分被HEVⅠ型或 /和Ⅱ型的诊断试剂排除的急性非戊肝病人血清对HEVⅣ型的多肽呈阳性反应。结论 为早期诊断戊肝病毒的感染 ,其诊断试剂应含有HEVⅣ型的ORF2N
Objective To express the entire ORF3 and four overlapping regions of ORF2 from HEV genotype Ⅳ and to analyze their immunoreactivities. Methods The fragments O3, FB5, E4, F2 2 and E5 were ligated into the expression vector pThioHis respectively; The expressed polypeptides were purified by reverse phase, size exclusion, ion exchange and affinity chromatography. The immunoassays were developed with each of the purified polypeptides. The sera from acute hepatitis E and non E hepatitis patients were tested with the assays. Results Polypeptide FB5, from the N terminus of ORF2, showed the greatest immunoreactivity with sera from patients with acute hepatitis; Some sera, from patients provisionally diagnosed as acute non E hepatitis based on Genelabs anti HEV IgG assay with antigens were from HEV genotype Ⅰ and Ⅱ, were positive for anti HEV IgG with the assays based on recombinant polypeptides from HEV genotype Ⅳ. This result indicates that the assays based on genotypes Ⅰ and Ⅱ alone are inadequate for detecting the HEV infection in China. Conclusion In order to diagnose the HEV infection early and correctly, the antigens used in the assays should include those from N terminus of ORF2 and from HEV genotype Ⅳ. [
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2002年第3期257-261,共5页
Chinese Journal of Microbiology and Immunology
基金
英国WellcomeTrust基金资助课题
