Lipscomb Jonathan T, Switzer William M, Li Jin-fen, Masciotra Silvina, Owen S Michele, Johnson Jeffrey A
Division of HIV/AIDS Prevention, Centers for Disease Control and Prevention, Atlanta, Georgia.
J Infect Dis. 2014 Dec 1;210(11):1827-37. doi: 10.1093/infdis/jiu333. Epub 2014 Jun 12.
Drug resistance mutations (DRMs) can serve as distinct, nonpolymorphic markers for evaluating diversity of expressed HIV-1. We screened for DRMs during early-acute viremia and examined the diversity in reverse transcriptase (RT) relative to envelope (env) in cases of transmitted drug resistance.
We evaluated 111 longitudinal plasma samples collected every 2-7 days from 15 individuals who seroconverted for HIV-1 infection in 1994-2000. The samples were screened with sensitive polymerase chain reaction assays for the commonly transmitted M41L and K70R mutations and for K65R, which was undetected by bulk sequencing. Mutation-positive samples were further characterized by clonal sequencing of RT and env V1-V3.
Drug resistance mutations were detected in 4 of 15 seroconverters at 5-50 days of viral nucleic acid expression; most mutations disappeared about the time of seroconversion. Clonal sequencing verified low-level K65R at frequencies of 0.4%-4.9%. In each case, K65R coexisted unlinked with variants carrying 2-5 thymidine analog mutations at frequencies of 1.6%-23.0%. In one seroconverter, variants with M184V and nonnucleoside RT inhibitor mutations were also identified at first RNA expression. Each seroconverter displayed a homogeneous V1-V3 env population.
Reverse-transcriptase DRMs demonstrate that the breadth of variants in transmission may be greater than what is reflected in envelope sequences.
耐药性突变（DRMs）可作为评估表达的HIV-1多样性的独特、非多态性标记。我们在急性早期病毒血症期间筛查了DRMs，并研究了传播性耐药情况下逆转录酶（RT）相对于包膜（env）的多样性。
我们评估了1994年至2000年期间15名HIV-1感染血清转化者每2至7天收集的111份纵向血浆样本。用敏感的聚合酶链反应检测法对样本进行筛查，以检测常见传播的M41L和K70R突变以及通过批量测序未检测到的K65R突变。对突变阳性样本通过RT和env V1-V3的克隆测序进行进一步特征分析。
在15名血清转化者中的4名在病毒核酸表达的5至50天检测到耐药性突变；大多数突变在血清转化时消失。克隆测序验证了低水平的K65R，频率为0.4%至4.9%。在每种情况下，K65R与携带2至5个胸苷类似物突变的变体不连锁共存，频率为1.6%至23.0%。在一名血清转化者中，在首次RNA表达时也鉴定出携带M184V和非核苷类逆转录酶抑制剂突变的变体。每名血清转化者均显示出均匀的V1-V3 env群体。
逆转录酶DRMs表明，传播中变体的广度可能大于包膜序列所反映的广度。
