Low-frequency HIV-1 Drug Resistance in Primary HIV-1 Infection

公共卫生监测项目使用共识测序（基因分型）估计，至少有十分之一， 在美国，抗逆转录病毒（ARV）初治的HIV-1感染者获得了耐药性HIV-1。 因此，指南建议在进入护理时进行耐药性测试。然而，共识 测序不能检测病毒群体中低于10-50%水平的低频变体。的 寡核苷酸连接测定（奥拉）是一种更灵敏的测定，其可以检测在低至 5%的病毒准种。在入组的原发性HIV-1感染受试者中进行的一项初步研究中， 在华盛顿大学原发性感染诊所（PIC），共有测序检测到 100名受试者中有6%的HIV-1耐药，奥拉在94名受试者中有28名（30%）检测到低频突变 受试者不具有通过共有测序鉴定的突变。 我们建议研究将使用奥拉，以解决有关的传输和随后的问题， HIV-1耐药性的后果。在目标1中，我们将研究ARV初治PIC受试者， 检测持续时间（“持久性”）和随着时间的推移检测到的HIV-1耐药性水平 外周血单核细胞（PBMC）、血液和精浆。在目标2中，我们将研究PIC受试者 启动ARV治疗并使用奥拉确定是否可以在PBMC中检测到其他突变 在成功治疗期间，由于选择了传播的低频耐药突变或 发展新的突变。在目标3中，我们将使用奥拉比较PIC中的HIV-1耐药模式 受试者及其来源伴侣，以确定HIV-1耐药性是否影响“传播适应性”。 这些新的研究将拓宽我们对低血糖的自然史和临床影响的理解， 艾滋病毒-1耐药性的频率，并为艾滋病毒感染者的检测和治疗提供指导。如果 更敏感的HIV-1耐药性检测将被批准用于临床护理， 了解低频突变的相关性，初始复杂性的潜在增加， 抗逆转录病毒治疗方案和随后患者依从性的降低可能会矛盾地增加 耐药性最后，来自伴侣对的经验数据将产生关于HIV-1相关性的信息。 传播，可以纳入未来的耐药性人口动态模型。 这些模型将估计由ARV传播的HIV-1耐药性的总体比例， 初次感染HIV-1的未接受过抗逆转录病毒治疗的来源伴侣与已确诊的 HIV-1感染。这些数据可用于设计针对这些人群的公共卫生干预措施， 减少HIV-1耐药性传播。

Public health surveillance programs using consensus sequencing (genotyping) estimate that at least one in ten antiretroviral (ARV)-na¿ve persons infected with HIV-1 in the United States acquires drug resistant HIV-1. Guidelines therefore recommend resistance testing at the time of entry into care. However, consensus sequencing cannot detect low-frequency variants at levels below 10-50% of the viral population. The oligonucleotide ligation assay (OLA) is a more-sensitive assay that can detect mutations occurring in as little as 5% of the viral quasi-species. In a pilot study conducted among subjects with primary HIV-1 infection enrolled at the University of Washington Primary Infection Clinic (PIC), consensus sequencing detected transmitted HIV-1 drug resistance in 6% of 100 subjects, and OLA detected low-frequency mutations in 28 (30%) of 94 subjects who did not have mutations identified by consensus sequencing. We propose studies that will use OLA to address questions pertaining to the transmission and subsequent consequences of HIV-1 drug resistance. In Aim #1, we will study ARV-na¿ve PIC subjects to compare the duration of detection ("persistence") and level of detection of transmitted HIV-1 drug resistance over time in peripheral blood mononuclear cells (PBMCs), blood and seminal plasma. In Aim #2, we will study PIC subjects initiating ARV therapy and use OLA to determine whether additional mutations can be detected in PBMCs during successful treatment due to the selection of transmitted low-frequency drug resistance mutations or the development of new mutations. In Aim #3, we will use OLA to compare HIV-1 drug resistance patterns in PIC subjects and their source partners to determine whether HIV-1 drug resistance impacts "transmission fitness". These novel investigations would broaden our understanding of the natural history and clinical impact of low- frequency HIV-1 drug resistance and inform guidelines for the testing and treatment of HIV-infected persons. If more-sensitive HIV-1 drug resistance assays were to be endorsed for clinical care before there is a full understanding of the relevance of low-frequency mutations, the potential increase in the complexity of initial ARV regimens and subsequent reduction in patient adherence could paradoxically increase the prevalence of drug resistance. Finally, empiric data from partner-pairs will generate information on correlates of HIV-1 transmission that could be incorporated into future models of the population dynamics of drug resistance. These models would estimate the overall proportion of HIV-1 drug resistance that is transmitted from ARV- na¿ve source partners with primary HIV-1 infection versus ARV-experienced source partners with established HIV-1 infection. This data could be used to design public health interventions targeted to these populations to reduce the spread of transmitted HIV-1 drug resistance.