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首页> 外文期刊>International journal of infectious diseases : >Comparison of susceptibility of HIV-1 variants to antiretroviral drugs by genotypic and recombinant virus phenotypic analyses
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Comparison of susceptibility of HIV-1 variants to antiretroviral drugs by genotypic and recombinant virus phenotypic analyses

机译：通过基因型和重组病毒表型分析比较HIV-1变体对抗逆转录病毒药物的敏感性

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Objective: This study utilized genotypic and in-house recombinant virus phenotypic assays to examine HIV-1 variant susceptibility to antiretroviral (ARV) drugs; comparisons were made between the analyses. Methods: A nested PCR was employed to amplify the HIV-1 gag-pol gene, which comprised the entire PR gene (codons 1-99) and the former RT gene (codons 1-312). Genetic resistance was determined by submitting the sequences to the Stanford University Network HIV-1 Database. Phenotypic susceptibilities to six ARV drugs were measured using a high-throughput, multi-cycle, recombinant virus phenotypic assay. Results were expressed in terms of the IC"5"0 (half maximal inhibitory concentration) and fold-change values. The relationship between phenotypic drug resistance and genetic polymorphisms was determined. Results: Nineteen fragment sequences for which recombinant viruses were successfully constructed were translated and compared with the consensus B sequences in the Stanford University Network HIV-1 Database. No recognizable genotypic resistance-associated mutations were noted, except in one sample. Each homologous replication-competent recombinant viral fold-change in the presence of six ARV drugs used widely in China was measured. According to the clinical and statistical criteria, 16 of the 19 samples were susceptible to the six drugs tested. The majority of phenotypic and genotypic results obtained were in agreement, with a concordance rate of 97.4%. Both phenotypic and genotypic assays suggested that sample HN2009001 was resistant to all drugs tested. All phenotypic and genotypic results obtained regarding the susceptibility of the 19 recombinant viruses to nucleoside reverse transcriptase inhibitors (NRTIs) were in agreement. With regard to the genotypic results for the non-nucleoside reverse transcriptase inhibitors (NNRTIs), 7.9% (3/38) were inconsistent with the phenotypic results. Conclusions: The in-house recombinant virus phenotypic assay was able to provide a straightforward quantitative assessment of resistance. In most cases, the genotypic and novel phenotypic assays yielded similar results. The disparity in HIV-1 susceptibility indicates a need to further investigate the clinical outcomes of antiretroviral therapy in certain individuals.
机译：目的：本研究利用基因型和内部重组病毒表型分析方法检测HIV-1变异体对抗逆转录病毒药物的敏感性。分析之间进行了比较。方法：采用巢式PCR扩增HIV-1 gag-pol基因，该基因由完整的PR基因（密码子1-99）和以前的RT基因（密码子1-312）组成。通过将序列提交斯坦福大学网络HIV-1数据库来确定遗传抗性。使用高通量，多周期，重组病毒表型测定法测量了对六种ARV药物的表型敏感性。结果以IC“ 5” 0（最大抑制浓度的一半）和倍数变化值表示。确定了表型耐药性与遗传多态性之间的关系。结果：成功构建重组病毒的19个片段序列被翻译，并与斯坦福大学网络HIV-1数据库中的共有B序列进行了比较。除一个样品外，未发现可识别的与基因型抗性相关的突变。在中国广泛使用的六种抗逆转录病毒药物存在下，对每种具有同源复制能力的重组病毒倍数进行了测量。根据临床和统计标准，在19个样品中有16个对六种药物敏感。获得的大多数表型和基因型结果一致，一致性率为97.4％。表型和基因型分析均表明样品HN2009001对所有测试的药物均具有抗性。关于19种重组病毒对核苷逆转录酶抑制剂（NRTIs）的敏感性，获得的所有表型和基因型结果都一致。关于非核苷类逆转录酶抑制剂（NNRTIs）的基因型结果，7.9％（3/38）与表型结果不一致。结论：内部重组病毒表型测定能够提供直接的耐药性定量评估。在大多数情况下，基因型和新的表型测定产生相似的结果。 HIV-1易感性差异表明需要进一步研究某些个体中抗逆转录病毒疗法的临床结果。

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