摘要
目的探讨不同基因型和亚型戊型肝炎病毒(HEV)ORF2重组蛋白p166用于抗体检测的价值,为研发准确可靠的戊型肝炎诊断试剂提供新的途径。方法用等浓度的不同基因型和亚型HEV p166作为包被抗原,对血清标本进行酶联免疫吸附试验测定。用HEV多基因型通用性引物逆转录套式PCR(RT-nPCR)扩增标本中HEV RNA,并测序、分型。结果8种不同p166抗原对30份健康献血者血清无抗原性,对182份来自世界不同国家和地区的已知HEV抗体阳性血清和7份HEV实验感染动物血清标本均呈阳性反应,但所得血清抗体滴度的高低与所用抗原的基因型有明显关系。 RT-nPCR检测的50份中国血清标本中,19份阳性,基因分型均为Ⅳ型,与Ⅳ型中国株p166抗原反应最好。而以同属于第Ⅲ基因型的猪HEV新西兰株和人HEV美国株重组p166检测血清标本,两者结果差异无统计学意义。以多基因型p166混合抗原建立的ELISA抗体检测法与两种市售试剂盒比较, 前者敏感性高,特异性好。结论不同基因型和亚型的。HEV重组蛋白p166对不同血清标本HEV抗体检测的敏感性高低不同,因此多基因型和亚型p166的混合抗原是HEV抗体检测的最佳抗原。
Objective To evaluate the application of p166 recombinant proteins derived from different genotypes and subtypes of hepatitis E virus （HEV） in anti-HEV antibody detection. Methods Anti-HEN antibodies were tested by indirect ELISA. RT-nPCR based on universal primers for multiple HEV genotypes was used to detect HEV RNA. Results No antigenicity of HEV was found in 30 serum samples obtained from normal blood donors. Positive reactions were observed in 182 serum specimens obtained from patients with acute hepatitis E and in 7 serum samples from experimenfly HEV-infeeted animals. Nineteen of 50 patient serum samples collected in China were RT-nPCR positive. All the positive isolates were clustered into HEV genotype Ⅳ. Moreover, titers of the anti-HEN antibody were associated with the genotypes of the antigens used in the ELISA detection. Comparing with two commercial ELISA kits, the method established in this study based on a mixture of multiple-genotype and subtype p166 antigens showed much higher sensitivity and specificity for anti-HEV antibody detection. The mixture of the multi-genotype and subtype HEV p166s is the best antigen apphed for HEV immunodiagnostics.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第4期369-374,共6页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金(30271231
30271212)江苏省自然科学基金(BK2002053)江苏省卫生厅医学科技发展基金(H200115) 教育部留学回国人员科研启动基金(2004-176)
