PDF(Pubmed)
Abstract：
We report a case of incomplete HIV-1 suppression on a dolutegravir, lamivudine, and abacavir single-tablet regimen with the emergence of the H51Y and G118R integrase resistance mutations.
Integrase sequencing was performed retrospectively by Sanger and next-generation sequencing. Rates of emergence and decline of resistance mutations were calculated using next-generation sequencing data. Dolutegravir plasma concentrations were measured by ultra-performance liquid chromatography-tandem mass spectrometry. The effects of H51Y and G118R on infectivity, fitness, and susceptibility to dolutegravir were quantified using cell-based assays.
During periods of non-adherence to treatment, mutations were retrospectively documented only by next-generation sequencing. Misdiagnosis by Sanger sequencing was caused by the rapid decline of mutant strains within the retroviral population. This observation was also true for a M184V lamivudine-resistant reverse transcriptase mutation found in association with integrase mutations on single HIV genomes. Resistance rebound upon treatment re-initiation was swift (>8000 copies per day). Next-generation sequencing indicated cumulative adherence to treatment. Compared to WT HIV-1, relative infectivity was 73%, 38%, and 43%; relative fitness was 100%, 35%, and 10% for H51Y, G118R, and H51Y+G118R viruses, respectively. H51Y did not change the susceptibility to dolutegravir, but G188R and H51Y+G118R conferred 7- and 28-fold resistance, respectively.
This case illustrates how poorly-fit drug-resistant viruses wax and wane alongside erratic treatment adherence and are easily misdiagnosed by Sanger sequencing. We recommend next-generation sequencing to improve the clinical management of incomplete virological suppression with dolutegravir.
Integrase sequencing was performed retrospectively by Sanger and next-generation sequencing. Rates of emergence and decline of resistance mutations were calculated using next-generation sequencing data. Dolutegravir plasma concentrations were measured by ultra-performance liquid chromatography-tandem mass spectrometry. The effects of H51Y and G118R on infectivity, fitness, and susceptibility to dolutegravir were quantified using cell-based assays.
During periods of non-adherence to treatment, mutations were retrospectively documented only by next-generation sequencing. Misdiagnosis by Sanger sequencing was caused by the rapid decline of mutant strains within the retroviral population. This observation was also true for a M184V lamivudine-resistant reverse transcriptase mutation found in association with integrase mutations on single HIV genomes. Resistance rebound upon treatment re-initiation was swift (>8000 copies per day). Next-generation sequencing indicated cumulative adherence to treatment. Compared to WT HIV-1, relative infectivity was 73%, 38%, and 43%; relative fitness was 100%, 35%, and 10% for H51Y, G118R, and H51Y+G118R viruses, respectively. H51Y did not change the susceptibility to dolutegravir, but G188R and H51Y+G118R conferred 7- and 28-fold resistance, respectively.
This case illustrates how poorly-fit drug-resistant viruses wax and wane alongside erratic treatment adherence and are easily misdiagnosed by Sanger sequencing. We recommend next-generation sequencing to improve the clinical management of incomplete virological suppression with dolutegravir.
摘要：
目的：我们报告一例dolutegravir不完全抑制HIV-1，拉米夫定,和阿巴卡韦单片治疗方案伴随H51Y和G118R整合酶抗性突变的出现。
方法：通过Sanger和下一代测序进行整合酶测序。使用下一代测序数据计算抗性突变的出现率和下降率。通过超高效液相色谱-串联质谱法测量Dolutegravir血浆浓度。H51Y和G118R对感染性的影响，健身,和对dolutegravir的易感性使用基于细胞的测定进行定量。
结果：在不坚持治疗期间，仅通过下一代测序对突变进行回顾性记录.Sanger测序的误诊是由逆转录病毒种群中突变株的快速下降引起的。对于与单个HIV基因组上的整合酶突变相关的M184V拉米夫定抗性逆转录酶突变也是如此。治疗重新开始时的阻力反弹迅速（每天>8000份）。下一代测序表明对治疗的累积依从性。与WTHIV-1相比，相对感染性为73%，38%,43%；相对适应度为100%，35%,H51Y为10%，G118R,和H51Y+G118R病毒,分别。H51Y没有改变对dolutegravir的易感性，但是G188R和H51Y+G118R赋予了7倍和28倍阻力，分别。
结论：该案例说明了不良耐药病毒是如何随着治疗依从性的不稳定而出现的，并且容易被Sanger测序误诊。我们建议使用下一代测序来改善dolutegravir不完全病毒学抑制的临床管理。
方法：通过Sanger和下一代测序进行整合酶测序。使用下一代测序数据计算抗性突变的出现率和下降率。通过超高效液相色谱-串联质谱法测量Dolutegravir血浆浓度。H51Y和G118R对感染性的影响，健身,和对dolutegravir的易感性使用基于细胞的测定进行定量。
结果：在不坚持治疗期间，仅通过下一代测序对突变进行回顾性记录.Sanger测序的误诊是由逆转录病毒种群中突变株的快速下降引起的。对于与单个HIV基因组上的整合酶突变相关的M184V拉米夫定抗性逆转录酶突变也是如此。治疗重新开始时的阻力反弹迅速（每天>8000份）。下一代测序表明对治疗的累积依从性。与WTHIV-1相比，相对感染性为73%，38%,43%；相对适应度为100%，35%,H51Y为10%，G118R,和H51Y+G118R病毒,分别。H51Y没有改变对dolutegravir的易感性，但是G188R和H51Y+G118R赋予了7倍和28倍阻力，分别。
结论：该案例说明了不良耐药病毒是如何随着治疗依从性的不稳定而出现的，并且容易被Sanger测序误诊。我们建议使用下一代测序来改善dolutegravir不完全病毒学抑制的临床管理。
