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飞机、机场及污水处理厂废水中新冠病毒基因组RNA和亚基因组RNA 5'端持久性的差异

Variations in the persistence of 5'-end genomic and subgenomic SARS-CoV-2 RNAs in wastewater from aircraft, airports and wastewater treatment plants.

作者信息

Tang Man-Hung Eric, Bennedbaek Marc, Gunalan Vithiagaran, Qvesel Amanda Gammelby, Thorsen Theis Hass, Larsen Nicolai Balle, Rasmussen Lasse Dam, Krogsgaard Lene Wulff, Rasmussen Morten, Stegger Marc, Alexandersen Soren

机构信息

Department of Bacteria, Parasites and Fungi, Statens Serum Institut, Copenhagen, Denmark.

Department of Virus and Microbiological Special Diagnostics, Statens Serum Institut, Copenhagen, Denmark.

出版信息

Heliyon. 2024 Apr 16;10(9):e29703. doi: 10.1016/j.heliyon.2024.e29703. eCollection 2024 May 15.

Abstract

Wastewater sequencing has become a powerful supplement to clinical testing in monitoring SARS-CoV-2 infections in the post-COVID-19 pandemic era. While its applications in measuring the viral burden and main circulating lineages in the community have proved their efficacy, the variations in sequencing quality and coverage across the different regions of the SARS-CoV-2 genome are not well understood. Furthermore, it is unclear how different sample origins, viral extraction and concentration methods and environmental factors impact the reads sequenced from wastewater. Using high-coverage, amplicon-based, paired-end read sequencing of viral RNA extracted from wastewater collected directly from aircraft, pooled from different aircraft and airport buildings or from regular wastewater plants, we assessed the genome coverage across the sample groups with a focus on the 5'-end region covering the leader sequence and investigated whether it was possible to detect subgenomic RNA from viral material recovered from wastewater. We identified distinct patterns in the persistence of the different genomic regions across the different types of wastewaters and the existence of chimeric reads mapping to non-amplified regions. Our findings suggest that preservation of the 5'-end of the genome and the ability to detect subgenomic RNA reads, though highly susceptible to environment and sample processing conditions, may be indicative of the quality and amount of the viral RNA present in wastewater.

摘要

在新冠疫情后的时代,废水测序已成为监测新冠病毒(SARS-CoV-2)感染的临床检测的有力补充。虽然其在测量社区病毒载量和主要流行谱系方面的应用已证明其有效性,但对于新冠病毒基因组不同区域的测序质量和覆盖范围的差异,人们尚未完全了解。此外,不同的样本来源、病毒提取和浓缩方法以及环境因素如何影响从废水中测序得到的读数也不清楚。我们使用从直接从飞机收集的废水、不同飞机和机场建筑物汇集的废水或常规污水处理厂收集的废水中提取的病毒RNA进行高覆盖度、基于扩增子的双端读数测序,评估了各样本组的基因组覆盖范围,重点关注覆盖前导序列的5'端区域,并研究了是否有可能从废水中回收的病毒材料中检测到亚基因组RNA。我们在不同类型废水的不同基因组区域的持久性以及映射到未扩增区域的嵌合读数的存在中发现了明显的模式。我们的研究结果表明,基因组5'端的保存以及检测亚基因组RNA读数的能力,尽管对环境和样本处理条件高度敏感,但可能表明废水中存在的病毒RNA的质量和数量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd70/11061675/dd580361e40c/gr1.jpg

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