Abstract：
Molluscum contagiosum virus (MOCV) is an important human pathogen causing a high disease burden worldwide. It is the last exclusively human-infecting poxvirus still circulating in its natural reservoir-a valuable model of poxviral evolution. Unfortunately, MOCV remains neglected, and little is known about its evolutionary history and circulating genomic variants, especially in non-privileged countries. The design weaknesses of available MOCV detection/genotyping assays surfaced with recent accumulation of abundant sequence information: all existing MOCV assays fail at accurate genotyping and capturing sub-genotype level diversity. Because complete MOCV genome characterization is an expensive and labor-intensive task, it makes sense to prioritize samples for whole-genome sequencing by diversity triage screening. To meet this demand, we developed a novel assay for accurate MOCV detection and genotyping, and comprehensive sub-genotype qualification to the level of phylogenetic groups (PGs). The assay included a novel set of oligonucleotide primers and probes, and it was implemented using digital polymerase chain reaction (dPCR). It offers sensitive, specific, and accurate detection, genotyping (MOCV1-MOCV3), and PG qualification (PG1-6) of MOCV DNA from clinical samples. The novel dPCR assay is suitable for MOCV diversity triage screening and prioritization of samples for complete MOCV genome characterization.
摘要：
传染性软疣病毒（MOCV）是一种重要的人类病原体，在全球范围内引起很高的疾病负担。它是最后一种完全感染人类的痘病毒，仍然在其天然水库中循环，这是一种有价值的痘病毒进化模型。不幸的是,MOCV仍然被忽视，对它的进化史和循环基因组变异知之甚少，特别是在非特权国家。可用的MOCV检测/基因分型测定的设计弱点随着最近大量序列信息的积累而浮出水面:所有现有的MOCV测定在准确的基因分型和捕获亚基因型水平多样性方面都失败。因为完整的MOCV基因组表征是一项昂贵且劳动密集型的任务，通过多样性分类筛选对样本进行全基因组测序是有意义的。为了满足这一需求,我们开发了一种准确的MOCV检测和基因分型的新方法，以及对系统发育组（PGs）水平的综合子基因型鉴定。该试验包括一组新的寡核苷酸引物和探针，它是使用数字聚合酶链反应（dPCR）实现的。它提供了敏感，具体,和准确的检测，基因分型（MOCV1-MOCV3），和来自临床样品的MOCVDNA的PG鉴定(PG1-6)。新型dPCR测定适用于MOCV多样性分类筛选和样品的优先级排序，以实现完整的MOCV基因组表征。
