摘要
目的 研究输血后肝炎病人的基因型 ,探讨 HCV抵抗 IFN疗效的作用机制。方法 以RT- PCR从慢性丙肝病人血清中扩增一段 4 86 bp的 c DNA片段作目的基因 ,插入 PQE30的多克隆位点 ,构建重组质粒 PQE30 - NS5 A- 1U 1D。结果 应用限制酶酶切和测序分析 ,证实 PQE30 -NS5 A- 1U1D中克隆基因是 HCVJ的 NS5 A部分序列 ,核苷酸和氨基酸序列同源性均在 90 %以上 ,包含有干扰素敏感决定区 (ISDR)。结论 血清学抗体检测 NS5 A片段对 HCV诊断有独特的应用价值 。
Objective To study the genotype of a patient after infecting Hepatitis C virus and analyse the mechanism which HCV resists to effect of β IFN. Methods A gene fragment about 486 bp of NS5A of HCV was cloned by RT PCR and gene recombinant techniques from the serum of a chronic HCV infected patient. The fragment was inserted into PQE30 vector and constructed the recombined plasmid: PQE30 NS5A 1U1D. Results Identified by restriction enzyme digestion and DNA sequencing , the NS5A cDNA fragment shared over 96% nucleotide acid and amino acid homology with HCVJ, including the interfon sensitivity determining region (ISDR). Conclusions Detection of antibody to HCV NS5A has a unique value in HCV diagnosis and may be helpful for evaluation of anti HCV therapy.
出处
《疾病控制杂志》
2003年第2期103-105,共3页
Chinese Journal of Disease Control and Prevention
基金
国家自然科学基金 (编号 30 170 85 6 )
