丙型肝炎病毒NS5a区血清型特异性多肽的选择及应用  

Selection and application of serotype-specific synthetic peptides derived from hepatitis C virus NS5a region

作　　者：窦晓光[1] 韩晓霜[1] 李智伟[1] 冯国和[1] 

机构地区：[1]中国医科大学附属盛京医院感染病科,辽宁沈阳110004

出　　处：《诊断学理论与实践》2007年第3期213-217,共5页Journal of Diagnostics Concepts & Practice

摘　　要：目的:从HCV非结构5a区(NS5a)选出基因型特异性多肽,并应用酶免疫法(EIA)进行HCV血清分型。方法:根据HCV NS5a高免疫源区(2182～2343)的氨基酸序列,设计并合成305个特异性多肽,应用已知基因1～6型的抗-HCV阳性血清,通过EIA法检测每个多肽的抗原性,并用基因型特异性多肽进行血清分型,同时将其与序列分析法基因分型结果进行比较。结果:不同基因型间NS5a区氨基酸的同源性较低,相同基因型不同区段氨基酸的同源性也很低。对305个特异性多肽抗原性检测结果表明,主要线性抗原区位于氨基酸残基R3、R7和R9区。18个来自保守区的多肽可与不同基因型抗-HCV阳性血清反应;12个多肽具基因型特异性。血清基因分型结果与基因分型的结果高度一致。结论:HCV NS5a高免疫源区存在主要的线性抗原;来自高度保守区的多肽抗原无基因型特异性,可用于HCV抗体检测;基因型特异性多肽可用于HCV基因分型,特别适用于HCV RNA阴性患者。Objective To select the genotype-specific synthetic peptides derived from hepatitis C virus （HCV） non- structure 5a （NS5a） region and set up an EIA method for HCV serotyping with the peptides. Methods Three hundred and five peptides within NS5a region at positions 2 182-2 343 of HCV were synthesized. All the peptides＇antigenic reactivity were tested by enzyme immunoassay with 69 human sera with anti-HCV positive representing genotypes 1-6. Forty hepatitis C positive patient sera were serotyped using serotype specific, synthetic peptides and genotyped by sequencing analysis. Results The correspondence of amino acids in HCV NS5a region withamino acids in positions 2 182-2 343 was very low among different genotype peptides. Using 305 peptides, antigenic regions were located in R3, R7 and R9. Eighteen peptides from highly conserved regions representing genotypes 1 to 6 showed broad immunoreactivity with sera containing antibody to all HCV genotypes. Twelve unique peptides showed highly, genotype specific, reactivity with same sera. The serotyping results showed high agreement with sequencing analysis. Conclusions There are the major antigenic regions in HCV NS5a region. Peptides from highly conserved region show genotype independent, immunoreactivity, useful for anti- HCV antibody test. Peptides from highly variable region show genotype dependent immunoreactivity, useful for determining HCV serotype, especially for patients with previously undetectable HCV RNA.

关 键 词：合成多肽 基因型特异性 丙型肝炎病毒 非结构5a区 

分 类 号：R466.6[医药卫生—临床医学]