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丙型肝炎病毒不同基因型NS3蛋白的抗原异质性分析
引用本文:乔伟振,孟继鸿,戴星,李丽,翟理杰,耿全林,周镇先.丙型肝炎病毒不同基因型NS3蛋白的抗原异质性分析[J].中华微生物学和免疫学杂志,2005,25(1):10-14.
作者姓名:乔伟振  孟继鸿  戴星  李丽  翟理杰  耿全林  周镇先
作者单位:1. 214001,无锡市第一人民医院中心实验室
2. 210009,南京,东南大学医学院病原生物学与免疫学系
3. 东南大学医学院附属中大医院
4. 东南大学医学院附属南京第二医院
基金项目:江苏省社会发展基金资助课题 (编号 :BS990 3 1)
摘    要:目的探讨不同基因型丙型肝炎病毒(HCV)NS3蛋白的抗原特性及其用于抗-HCV检测的意义。方法分别构建和表达含有HCV1型和6型NS3基因片段的重组质粒和重组蛋白,以EIJSA法和Western blot分析不同基因型重组蛋白与已知抗-HCV阳性血清的抗原反应性。结果HCV1型和6型NS3重组蛋白氨基酸序列的同源性为83.2%;85份抗-HCV阳性血清以此不同基因型HCV NS3单片段抗原检测,阳性检出率分别为61.2%(1型)和58.8%(6型),其中有7份标本以NS3-1型抗原检测阴性,但可被NS3-6型抗原检出,反之,有9份血清以NS3-6型重组蛋白检测为阴性,而NS3.1型检测阳性;54份大学生体检血清和39份阴性质控血清以此两种抗原检测均为阴性。结论HCV1型和6型NS3重组蛋白存在抗原异质性,在发展HCV抗体检测试剂时需考虑加入不同基因型的NS3抗原。

关 键 词:NS3蛋白  抗原  重组蛋白  丙型肝炎病毒  阴性  HCV  基因型  氨基酸序列  片段  异质性
修稿时间:2004年7月26日

Antigenic heterogeneity of the hepatitis C virus NS3 proteins derived from different genotypes
QIAO Wei-zhen,MENG Ji-hong,DAI Xing,LI Li,ZHAI Li-jie,GENG Quan-lin,ZHOU Zhen-xian.Antigenic heterogeneity of the hepatitis C virus NS3 proteins derived from different genotypes[J].Chinese Journal of Microbiology and Immunology,2005,25(1):10-14.
Authors:QIAO Wei-zhen  MENG Ji-hong  DAI Xing  LI Li  ZHAI Li-jie  GENG Quan-lin  ZHOU Zhen-xian
Institution:QIAO Wei-zhen*,MENG Ji-hong,DAI Xing,LI Li,ZHAI Li-jie,GENG Quan-lin,ZHOU Zhen-xian. *Department of Microbiology and Immunology,Southeast University School of Medicine,Nanjing 210009,China
Abstract:Objective To characterize the antigenicity of the hepatitis C virus (HCV) NS3 recombinant proteins derived from different genotypes and to identify the significance of the proteins used for anti-HCV detection. Methods The recombinant plasmids of pET-NS3-1 and pET-NS3-6 were constructed with pET-28a-c vector and PCR products of HCV NS3 regions derived from HCV genotype 1 and 6 respectively. The antigenic heterogeneity of the HCV NS3 proteins was determined by ELISA and Western blot with the known anti-HCV-positive serum samples. Results The homology of the amino acid sequences of the HCV NS3 recombinant proteins was 83.2% between HCV genotype 1 and 6. The positive rates of anti-NS3 were 61.2% by using the NS3 antigen from HCV genotype 1 and 58.8% by using the antigen from genotype 6, respectively, among 85 known anti-HCV-positive specimens. However, 7 specimens detected with the antigen from genotype 1 were negative but positive when detected with the antigen from genotype 6. On the contrary, 9 specimens were negative with the HCV NS3 antigen from genotype 6 but were positive with the antigen from genotype 1. Fifty-four serum samples collected from normal undergraduates and 39 specimens from an anti-HCV-negative panel were both negative detected by both the NS3 antigens. Conclusion Antigenic heterogeneity of HCV NS3 proteins exists between HCV genotype 1 and 6 and has a profound effect on the anti-HCV detection. This observation should be taken into consideration in the further development of diagnostic assays for the detection of anti-HCV activity.
Keywords:Hepatitis C virus  NS3  Recombinant antigen  Enzyme-linked immunosorbent assay(ELISA)
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