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Updated: Jul 26, 2025

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Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
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开发一种基于片的测序方法,以应对全球出现的mopox病毒
Nicholas F G Chen1, Chrispin Chaguza1, Luc Gagne2
1Department of Epidemiology of Microbial Diseases, Yale School of Public Health, New Haven, Connecticut, United States of America.
PLoS biology
|June 13, 2023
概括
基于Amplicon的测序提供了一种敏感且具有成本效益的方法,用于对mopox病毒的全基因组测序,即使病毒载量低. 这种方法在2022年mopox疫情期间被证明是非常有效的,增强了基因组监测能力.
科学领域:
- 基因组监测是指基因组的监测.
- 病毒病原体的测序.
- 公共卫生准备 公共卫生准备
背景情况:
- 2022年mopox疫情突出了资源密集型元基因组测序的局限性,以快速识别病原体.
- 需要一种更敏感和更广泛的测序方法,特别是对于具有可变病毒载荷的样本.
- 现有的基于amplicon的测序方法,如PrimalSeq,已经适应了寨卡病毒和SARS-CoV-2等病毒.
研究的目的:
- 开发和验证一种基于安普利康的敏感测序方法,用于人类病毒 (mpox).
- 评估这种方法在各种临床标本和公共卫生实验室的有效性.
- 为快速基因组监测新出现的病原体提供一个具有成本效益和灵活性的工具.
主要方法:
- 开发基于PrimalScheme的原始基因组,用于人类病毒全基因组测序.
- 使用基于amplicon和metagenomic方法对临床标本进行比较测序.
- 为十个国际公共卫生实验室进行验证,分配了初级池的配额.
主要成果:
- 与元基因组学方法相比,基于AMPLCON的测序显示出更高的基因组覆盖率和更少的脱学率,特别是在具有更高PCR周期值 (Ct) 值的样本中.
- PCR Ct值与测序读数和基因组覆盖率相关,最佳结果低于31 Ct和100万读数.
- 国际实验室成功地在各种工作流程,样品类型和Ct值中实施了原料方案,证实了其广泛的适用性.
结论:
- 基于AMPlicon的测序提供了一个快速部署的,具有成本效益和灵活的方法,用于像mopox.com这样的新兴病原体的全基因组测序.
- 开发的初始化方案很容易适应现有的实验室工作流程,包括用于SARS-CoV-2监测的工作流程.
- 这种方法显著提高了国家和国际快速基因组监测和疫情应对能力.
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