New antivirals are required to prevent rising antimicrobial resistance from replication inhibitors. The aim of this study was to analyse the range of emerging mutations in herpesvirus by whole genome deep sequencing. We tested human herpesvirus 6 treatment with novel antiviral K21, where evidence indicated distinct effects on virus envelope proteins. We treated BACmid cloned virus in order to analyse mechanisms and candidate targets for resistance. Illumina based next generation sequencing technology enabled analyses of mutations in 85 genes to depths of 10,000 per base detecting low prevalent minority variants (<1%). After four passages in tissue culture the untreated virus accumulated mutations in infected cells giving an emerging mixed population (45–73%) of non-synonymous SNPs in six genes including two envelope glycoproteins. Strikingly, treatment with K21 did not accumulate the passage mutations; instead a high frequency mutation was selected in envelope protein gQ2, part of the gH/gL complex essential for herpesvirus infection. This introduced a stop codon encoding a truncation mutation previously observed in increased virion production. There was reduced detection of the glycoprotein complex in infected cells. This supports a novel pathway for K21 targeting virion envelopes distinct from replication inhibition.

需要新的抗病毒药来防止复制抑制剂引起的抗药性上升。这项研究的目的是通过全基因组深度测序分析疱疹病毒中新出现的突变范围。我们用新型抗病毒K21测试了人疱疹病毒6的治疗，证据表明对病毒包膜蛋白有明显的作用。我们分析了BACmid克隆病毒，以分析抗药性的机制和候选靶标。基于Illumina的下一代测序技术能够分析85个基因的突变，每个碱基深度达10,000个，从而检测出低流行性少数变异（<1％）。经过四次传代培养后，未经处理的病毒在受感染的细胞中积累了突变，从而在包括两个包膜糖蛋白在内的六个基因中出现了新兴的非同义SNP混合种群（45-73％）。惊人地 K21治疗未累积传代突变；而是在包膜蛋白gQ2中选择了高频突变，该蛋白是疱疹病毒感染必不可少的gH / gL复合体的一部分。这引入了编码先前在增加的病毒体产量中观察到的截短突变的终止密码子。减少了感染细胞中糖蛋白复合物的检测。这支持了不同于复制抑制的K21靶向病毒粒子包膜的新途径。减少了感染细胞中糖蛋白复合物的检测。这支持了不同于复制抑制的K21靶向病毒粒子包膜的新途径。减少了感染细胞中糖蛋白复合物的检测。这支持了不同于复制抑制的K21靶向病毒粒子包膜的新途径。