在海上军事环境中部署 SARS-CoV-2 全基因组下一代测序。

IF 1.4 4区 医学 Q2 MEDICINE, GENERAL & INTERNAL Bmj Military Health Pub Date : 2024-12-11 DOI:10.1136/military-2022-002296
Andrew Bosworth, J Robson, B Lawrence, A L Casey, A Fair, S Khanam, C Hudson, M K O'Shea
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引用次数: 0

摘要

背景:SARS-CoV-2 可在海上平台迅速传播。据报道,在海上的军舰上爆发过几次 SARS-CoV-2 疫情,因为在近距离的生活和工作环境中容易传播。感染控制措施的核心内容,如社会隔离、病人隔离和接触者检疫,都极难实施。SARS-CoV-2 的全基因组测序(WGS)促进了对疫情的流行病学调查,通过确定传播模式、感染群和指导控制措施,对疫情的实时管理产生了影响。我们认为这种能力可以减轻 SARS-CoV-2 在海上环境中的影响:方法:我们利用微型纳米孔测序技术,在皇家舰队辅助船 ARGUS 号上建立了海上 SARS-CoV-2 WGS。目标包括设计一个简化的方案,只需最少的试剂和处理步骤,使用可在有限空间内使用的小型化设备,以及简化和独立的数据分析能力,以实现快速的原位数据采集和解读:结果:在海上检测了 11 份具有 SARS-CoV-2 状态的临床盲样。病毒 RNA 提取和 ARTIC 测序文库制备完成后,进行了反转录和 ARTIC PCR 扩增。随后使用牛津纳米孔 MinION Mk1B 对样本进行条形码编码和测序。使用离线版 MinKNOW 软件,然后使用 CLC Genomics Workbench 进行下游分析,以进行变异识别和系统发生树构建。所有样本都被正确分类,并确定了亲缘关系:结论:在海上军事环境中,由于后方支援有限,建立小规模测序能力以进行 SARS-CoV-2 WGS 是可行的。这项概念验证研究强调了将来在海上和陆地军事环境中部署这种技术的潜力,以提供有意义的临床数据,协助疫情调查。
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Deployment of whole genome next-generation sequencing of SARS-CoV-2 in a military maritime setting.

Background: SARS-CoV-2 can spread rapidly on maritime platforms. Several outbreaks of SARS-CoV-2 have been reported on warships at sea, where transmission is facilitated by living and working in close quarters. Core components of infection control measures such as social distancing, patient isolation and quarantine of exposed persons are extremely difficult to implement. Whole genome sequencing (WGS) of SARS-CoV-2 has facilitated epidemiological investigations of outbreaks, impacting on outbreak management in real time by identifying transmission patterns, clusters of infection and guiding control measures. We suggest such a capability could mitigate against the impact of SARS-CoV-2 in maritime settings.

Methods: We set out to establish SARS-CoV-2 WGS using miniaturised nanopore sequencing technology aboard the Royal Fleet Auxiliary ARGUS while at sea. Objectives included designing a simplified protocol requiring minimal reagents and processing steps, the use of miniaturised equipment compatible for use in limited space, and a streamlined and standalone data analysis capability to allow rapid in situ data acquisition and interpretation.

Results: Eleven clinical samples with blinded SARS-CoV-2 status were tested at sea. Following viral RNA extraction and ARTIC sequencing library preparation, reverse transcription and ARTIC PCR-tiling were performed. Samples were subsequently barcoded and sequenced using the Oxford Nanopore MinION Mk1B. An offline version of the MinKNOW software was used followed by CLC Genomics Workbench for downstream analysis for variant identification and phylogenetic tree construction. All samples were correctly classified, and relatedness identified.

Conclusions: It is feasible to establish a small footprint sequencing capability to conduct SARS-CoV-2 WGS in a military maritime environment at sea with limited access to reach-back support. This proof-of-concept study has highlighted the potential of deploying such technology in the future to military environments, both maritime and land-based, to provide meaningful clinical data to aid outbreak investigations.

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来源期刊
Bmj Military Health
Bmj Military Health MEDICINE, GENERAL & INTERNAL-
CiteScore
3.10
自引率
20.00%
发文量
116
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