González-Peralta R P, Qian K, She J Y, Davis G L, Ohno T, Mizokami M, Lau J Y
Department of Medicine, University of Florida, Gainesville 32610, USA.
J Med Virol. 1996 Jul;49(3):242-7. doi: 10.1002/(SICI)1096-9071(199607)49:3<242::AID-JMV14>3.0.CO;2-E.
To determine the clinical significance of viral quasispecies heterogeneity, 59 patients with chronic hepatitis C were studied using singlestranded conformational polymorphism (SSCP) analysis of the HCV E2 hypervariable region 1 (HVR1); of these, 48 were subsequently treated with interferon-alpha. The SSCP method was validated using clones of known nucleotide sequence. HVR1 was amplified in 54 of 59 (92%) patients. The median number of SSCP bands per sample was 6 (range: 2-12). Increased quasispecies heterogeneity correlated with the estimated duration of HCV infection (P < 0.05), parenteral-acquired HCV infection (vs. sporadic, P < 0.05), serum HCV RNA levels (P < 0.05), and HCV genotype 1 infection (P < 0.05), but not with age, serum AST, ALT, or Knodell score. Patients who had complete and sustained response to interferon-alpha (n = 11) had lower pre-treatment quasispecies heterogeneity compared to patients who had complete response with relapse (n = 18, P < 0.05) or no complete response (n = 16, P < 0.01). However, multivariate analysis revealed that HCV viremia was a stronger predictor of response to interferon-alpha. These findings indicate that the estimated duration of HCV carriage, serum HCV RNA levels, and HCV type 1 are important determinants for the evolution of HCV quasispecies heterogeneity; and that increased HCV quasispecies heterogeneity is another marker associated with a poor subsequent response to interferon-alpha.
为了确定病毒准种异质性的临床意义，我们使用丙型肝炎病毒（HCV）E2高变区1（HVR1）的单链构象多态性（SSCP）分析对59例慢性丙型肝炎患者进行了研究；其中48例随后接受了α干扰素治疗。使用已知核苷酸序列的克隆对SSCP方法进行了验证。59例患者中有54例（92%）扩增出了HVR1。每个样本的SSCP条带中位数为6条（范围：2 - 12条）。准种异质性增加与HCV感染的估计持续时间（P < 0.05）、经肠外途径获得的HCV感染（与散发感染相比，P < 0.05）、血清HCV RNA水平（P < 0.05）以及HCV 1型感染（P < 0.05）相关，但与年龄、血清谷草转氨酶（AST）、谷丙转氨酶（ALT）或Knodell评分无关。与出现完全应答但复发的患者（n = 18, P < 0.05）或未出现完全应答的患者（n = 16, P < 0.01）相比，对α干扰素产生完全且持续应答的患者（n = 11）治疗前的准种异质性较低。然而，多变量分析显示HCV病毒血症是对α干扰素应答的更强预测指标。这些发现表明，HCV携带的估计持续时间、血清HCV RNA水平以及HCV 1型是HCV准种异质性演变的重要决定因素；并且HCV准种异质性增加是另一个与随后对α干扰素应答不佳相关的标志物。
