丙型肝炎病毒感染者F基因区变异临床意义的初步研究

目的探讨丙型肝炎病毒（hepatitis C virus,HCV）F基因区变异的临床意义。方法以78例未治疗慢性丙型肝炎患者为研究对象,采用单链构象多态性分析对HCV-F基因区进行准种分析,实时荧光定量聚合酶链反应对HCV RNA进行定量检测,利用酶联免疫吸附试验分别对患者血清中的抗F蛋白抗体、Th1（IL-12,IFN-γ）和Th2（IL-5,IL-10）细胞因子水平进行检测分析。结果 78例慢性丙型肝炎患者中有46例为抗HCV-F抗体阳性,阳性率为59%。HCV1b亚型患者F基因区准种数目高于非HCV1b亚型（P=0.002）。F基因区SSCP条带数与HCV RNA载量及ALT/AST水平呈正相关（均有P〈0.05）。另外,与F基因准种数量≤2的患者相比,F基因准种数量〉2的患者血清中,Th1细胞因子IL-12、IFN-γ水平降低（P=0.023,P=0.010）,Th2细胞因子IL-5、IL-10水平显著升高（P=0.024,P=0.012）。结论 HCV-F基因区准种多样性与病毒复制、肝损伤和机体Th2优势应答有关,具有一定的临床意义。

Study on the diversity and its clinical significance of HCV gene F region

Objective To explore the relationship between the HCV F region quasispecies and the clinical significance. Methods HCV quasispecies from 78 un-treated patients were detected by the single-strand conformation polymorphism（ SSCP） analysis. The HCV RNA was quantitatively analysed by the real-time fluorescence quantitative polymerase chain reaction. The antibody of HCV F protein and Th1（ IL-12,IFN-γ） or Th2（ IL-5,IL-10） cytokines were detected by the enzyme-linked immunosorbent assay. Results The positive rate of anti-F antibody in chronic HCV patients was 59% and the number of SSCP bands of patients with HCV 1b subtype was significantly higher than that of patients with non-HCV 1b subtype（ P = 0. 002）. Moreover,the number of quasispecies was associated with the loads of HCV RNA or levels of serum ALT/AST in chronic patients（ all P 0. 05）. In addition,the levels of Th1（ IL-12,IFN-γ） cytokines secreted in the patients with the number of F quasispecies 2 were lower than those of patients with F quasispecies ≤2（ P = 0. 023; P = 0. 010）,and the Th2（ IL-5,IL-10） cytokines in the patients with the number of F quasispecies 2 were higher than those with F quasispecies≤2（ P = 0. 024; P = 0. 012）. Conclusions The number of HCV F region quasispecies correlates with the viral replication,liver damage and Th2 dominant response.