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应用鸟枪法宏基因组测序和靶向序列捕获技术检测荷兰-德国边境地区循环猪病毒。

Application of shotgun metagenomics sequencing and targeted sequence capture to detect circulating porcine viruses in the Dutch-German border region.

机构信息

Department of Medical Microbiology and Infection Prevention, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.

Institute of Medical Microbiology and Hygiene, University of Tübingen, Tübingen, Germany.

出版信息

Transbound Emerg Dis. 2022 Jul;69(4):2306-2319. doi: 10.1111/tbed.14249. Epub 2021 Aug 28.

Abstract

Porcine viruses have been emerging in recent decades, threatening animal and human health, as well as economic stability for pig farmers worldwide. Next-generation sequencing (NGS) can detect and characterize known and unknown viruses but has limited sensitivity when an unbiased approach, such as shotgun metagenomics sequencing, is used. To increase the sensitivity of NGS for the detection of viruses, we applied and evaluated a broad viral targeted sequence capture (TSC) panel and compared it to an unbiased shotgun metagenomic approach. A cohort of 36 pooled porcine nasal swab and blood serum samples collected from both sides of the Dutch-German border region were evaluated. Overall, we detected 46 different viral species using TSC, compared to 40 viral species with a shotgun metagenomics approach. Furthermore, we performed phylogenetic analysis on recovered influenza A virus (FLUAV) genomes from Germany and revealed a close similarity to a zoonotic influenza strain previously detected in the Netherlands. Although TSC introduced coverage bias within the detected viruses, it improved sensitivity, genome sequence depth and contig length. In-depth characterization of the swine virome, coupled with developing new enrichment techniques, can play a crucial role in the surveillance of circulating porcine viruses and emerging zoonotic pathogens.

摘要

近年来,猪病毒不断出现,威胁着动物和人类的健康,也影响着全球养猪户的经济稳定。下一代测序(NGS)可以检测和描述已知和未知的病毒,但当使用无偏方法(如 shotgun 宏基因组测序)时,其灵敏度有限。为了提高 NGS 检测病毒的灵敏度,我们应用并评估了广泛的靶向病毒序列捕获(TSC)面板,并将其与无偏 shotgun 宏基因组方法进行了比较。我们评估了来自荷兰-德国边境地区两侧的 36 个混合猪鼻拭子和血清样本。总的来说,我们使用 TSC 检测到了 46 种不同的病毒,而使用 shotgun 宏基因组方法检测到了 40 种病毒。此外,我们对从德国回收的甲型流感病毒(FLUAV)基因组进行了系统发育分析,结果表明与之前在荷兰检测到的一种人畜共患流感株密切相似。尽管 TSC 在检测到的病毒中引入了覆盖偏差,但它提高了灵敏度、基因组序列深度和连续序列长度。深入了解猪的病毒组,并结合开发新的富集技术,可以在监测循环猪病毒和新发人畜共患病原体方面发挥关键作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9448/9540031/37724ff2f280/TBED-69-2306-g002.jpg

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