摘要
目的 检测不同基因型丙型肝炎病毒 (HCV)非结构 5a区 (NS5a)高免疫原区短链多肽的抗原性 ,确定该区的主要线性抗原表位并探讨基因异质性与免疫原性间的关系。方法 设计并合成特异性多肽 ;DNAStar软件对多肽氨基酸同源性进行比较 ;间接ELISA法检测多肽的抗原性。结果多肽氨基酸的同源性在不同区及不同基因型间变化较大。氨基酸残基 2 2 12～ 2 2 4 1、2 2 72～ 2 30 1和2 30 2～ 2 331的多肽抗原性最强。 18个来自保守区的多肽可以与不同基因型抗 HCV阳性血清起反应 (阳性率高达 96 % ) ;而来自不同基因型间氨基酸序列保守性较低区的多肽抗原性与同基因血清的反应性较高。结论 HCVNS5a高免疫原区主要的线性抗原位于氨基酸残基 2 2 12～ 2 2 4 1、2 2 72～2 30 1和 2 30 2～ 2 331的位置 ;人工合成的多肽可以有效地用于检测抗 HCV抗体 ;
Objective To identify the location of major immnodominant antigenic region and study the relationship between the gene heterogeneity and immunoreactivity via detecting antigenic reactivity of synthetic peptides deriving from immunodominant region in different genotypes of hepatitis C virus (HCV) NS5a gene. Methods In total, 305 non-identical 30-mer long and overlapping by 15 aa peptides derived from HCV NS5a region from codon 2 182 to 2 343 among 45 unique published HCV sequences in GenBank corresponding to different genotype were designed and synthesized. The amino acid sequences of all peptides were compared with DNA Star software. The antigenic reactivity of those peptides was detected with indirect ELISA with both anti-HCV and anti-NS5 positive serum. Results The sequences showed highly conserved among HCV genotype in regions 2 272 -2 301 and 2 302-2 331 as compared to regions 2 212-2 241 and 2 257-2 286. The peptides basing on amino acid residues among 2 212-2 241,2 272-2 301 and 2 302-2 331 showed stronger immunoreactivity than any other peptides. Eighteen peptides derived from this region showed a broad immunoreactivity, 3 of them could react with 96% of anti-HCV positive sera. Whereas the immunoreactivity of the peptides derived from the region showing highly variable among HCV genotype was found to react more strongly with homologous-genotype sera. Conclusion The major linear antigenic region was located at amino acid residues among 2 212-2241,2 272-2 301 and 2 302- 2 331 ; the short synthetic peptide derived from NS5a region at position 2 212-2 241,2 272-2 301 and 2 302- 2 331 can be used for efficient detection of HCV antibody; some peptides showed genotype specific immuunoreactivity.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
北大核心
2004年第4期344-347,共4页
Chinese Journal of Experimental and Clinical Virology
