A novel digital PCR assay for detection and comprehensive characterization of Molluscum contagiosum virus genotypes MOCV1, MOCV2, and MOCV3 and recombinant lineages

IF 2.2 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS
Tomaž M. Zorec , Lucijan Skubic , Mario Poljak
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引用次数: 0

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.

一种新型数字 PCR 检测法,用于检测和全面鉴定传染性软疣病毒基因型 MOCV1、MOCV2 和 MOCV3 以及重组系。
传染性软疣病毒(MOCV)是一种重要的人类病原体,在全球范围内造成沉重的疾病负担。它是最后一种仍在其自然储库中循环的专门感染人类的痘病毒,是痘病毒进化的宝贵模型。遗憾的是,MOCV 仍被忽视,人们对其进化史和循环基因组变种知之甚少,尤其是在非特权国家。随着最近大量序列信息的积累,现有 MOCV 检测/基因分型检测方法在设计上的弱点逐渐浮出水面:所有现有的 MOCV 检测方法都无法进行准确的基因分型和捕捉亚基因型水平的多样性。由于完整的 MOCV 基因组特征描述是一项昂贵且劳动密集型的任务,因此通过多样性分流筛选来优先对样本进行全基因组测序是非常有意义的。为了满足这一需求,我们开发了一种新型检测方法,用于准确检测 MOCV 和进行基因分型,并对系统发生群(PGs)水平的亚基因型进行全面鉴定。该检测方法包括一套新型的寡核苷酸引物和探针,并采用数字聚合酶链式反应(dPCR)实现。它能对临床样本中的 MOCV DNA 进行灵敏、特异和准确的检测、基因分型(MOCV1-MOCV3)和 PG 鉴定(PG1-6)。这种新型的 dPCR 检测方法适用于 MOCV 多样性分流筛选,并能优先选择样本进行完整的 MOCV 基因组鉴定。
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来源期刊
CiteScore
5.80
自引率
0.00%
发文量
209
审稿时长
41 days
期刊介绍: The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.
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