使用牛津纳米孔技术开发和评估麻疹病毒下一代测序方法
Vanessa Zubach1, Gurasis Osahan1, Kurt Kolsun1
1Measles, mumps and rubella Unit, National Microbiology Laboratory, Public Health Agency of Canada, JC Wilt infectious Diseases Research Centre, Winnipeg, Manitoba, Canada.
Journal of clinical microbiology
|February 4, 2025
概括
这项研究表明,牛津纳米孔技术测序为麻疹病毒提供了准确的全基因组序列,增强了全球监测能力. 开发的协议确保了公共卫生响应的可靠数据,即使采样量很小.
科学领域:
- 病毒学和基因组学
- 生物信息学和计算生物学
- 公共卫生和流行病学
背景情况:
- 麻疹病毒 (MeV) 全基因组测序 (WGS) 对于流行病监测和疫情管理至关重要.
- 现有的测序方法可能昂贵且难以获得,限制了全球数据生成,特别是在资源有限的地区.
研究的目的:
- 使用牛津纳米孔技术 (ONT) 开发和评估MeV基因型D8 WGS的基准协议和生物信息管道.
- 评估ONT生成的MeV WGS的准确性,可靠性和实用性,用于麻疹监测.
主要方法:
- 开发一个使用1kb的安普利康进行MeV WGS的基板协议.
- 评估四个生物信息学管道参数 (Guppy简单和多拉多双重,具有不同的Q分数值).
- 使用参考基因组和32个麻疹病例样本的小组与已知的参考序列进行验证.
主要成果:
- 获得完整的MeV基因组低至10个副本/μL,检测到的错误在100个副本/μL.
- 过点 (Cp) 值≤31预测准确和完整的WGS生成.
- 与参考结果相比,GQ20参数产生了最完整 (75%) 和最相同 (84.4%) 的序列,具有很高的可重复性 (97.4%).
- 由ONT生成的WGS数据与与流行病相关联的疫情数据相结合.
结论:
- ONT测序平台提供了一个具有成本效益和便携性的解决方案,用于生成准确可靠的MeV WGS.
- 这项技术有可能在全球范围内显著扩大麻疹测序的可用性,改善公共卫生的及时数据生成.
- 开发的协议和管道适用于加强麻疹监测,特别是在序列数据有限的地区.
相关概念视频
03:00Next-generation Sequencing
87.2K
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
87.2K
03:21RNA-seq
9.8K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
9.8K