Middleton Tim, He Yupeng, Pilot-Matias Tami, Tripathi Rakesh, Lim Ben Hock, Roth Andrew, Chen Chih-Ming, Koev Gennadiy, Ng Teresa I, Krishnan Preethi, Pithawalla Ron, Mondal Rubina, Dekhtyar Tatyana, Lu Liangjun, Mo Hongmei, Kati Warren M, Molla Akhteruzzaman
Abbott Laboratories, Global Pharmaceutical Research and Development, Department R4CQ, AP52N, 200 Abbott Park Road, Abbott Park, IL 60064, USA.
J Virol Methods. 2007 Nov;145(2):137-45. doi: 10.1016/j.jviromet.2007.05.016. Epub 2007 Jun 29.
Hepatitis C virus (HCV) replicon-based shuttle vectors that permit phenotypes of NS5B polymerase genes from a large number of patient isolates to be rapidly assessed when transiently expressed in cultured cells were designed. When used to test responses to an inhibitor of HCV RNA-dependent RNA polymerase, IC(50) values for inhibition covered a several hundred-fold range among 47 patient samples tested. This observation highlights the variability that can be found by testing isolates derived from HCV-infected subjects. Partial suppression with a polymerase inhibitor of the most sensitive species permitted detection of minor quasispecies that were 7-200-fold more resistant than the bulk population in approximately half of the samples. Sequence analysis showed a wide range of amino acid changes not detected by conventional selection methods using laboratory-derived strains. This approach provides a means to assess variation in antiviral efficacy, and to predict possible responses in a clinical setting.
设计了基于丙型肝炎病毒（HCV）复制子的穿梭载体，当在培养细胞中瞬时表达时，可快速评估来自大量患者分离株的NS5B聚合酶基因的表型。当用于测试对HCV RNA依赖性RNA聚合酶抑制剂的反应时，在47个测试的患者样本中，抑制的IC（50）值覆盖了几百倍的范围。这一观察结果突出了通过测试源自HCV感染受试者的分离株所发现的变异性。在大约一半的样本中，用最敏感物种的聚合酶抑制剂进行部分抑制，可检测到比总体群体耐药性高7至200倍的次要准种。序列分析显示了使用实验室衍生菌株的传统选择方法未检测到的广泛氨基酸变化。这种方法提供了一种评估抗病毒疗效差异并预测临床环境中可能反应的手段。
