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使用纳米孔技术对口病病毒完整基因组进行普遍放大和测序

Andrew E Shaw¹, Kebaneilwe Lebani^2,3, Lina González Gordon⁴

¹The Pirbright Institute, Ash Road, Pirbright, Surrey, GU24 0NF, UK. andrew.shaw@pirbright.ac.uk.

|August 22, 2025

概括

使用纳米孔测序的新通用协议现在可以完全描述口病病毒 (FMDV) 基因组. 这种方法不依赖于血统,为全球追踪口疾疫情提供了至关重要的工具.

科学领域:

病毒学
基因组学
生物信息学

背景情况:

足口病病毒 (FMDV) 在两动物中引起严重的疫情.
基因组表征对于追踪口疾病的起源和传播至关重要.
现有的测序方法通常针对特定的FMDV谱系或仅针对片段.

研究的目的:

开发一个通用口病病毒 (FMDV) 测序协议.
创建一个生物信息管道来组装任何FMDV基因组.
为了实现血统无关的FMDV基因组测序.

主要方法:

全面多重RT-PCR在重叠的片段中放大了整个FMDV基因组.
使用MinION设备进行纳米孔测序.
采用了使用blastn和参考组件的生物信息管道.

主要成果:

开发了两种原始方案 (S_scheme和L_scheme),两种方案都能够生成FMDV基因组.
对于完整的基因组生成,L_scheme被证明更简单,更可靠,更具成本效益.
该方案在30种不同型号的口疫苗上得到了验证,并在乌干达进行了试验.

结论:

开发的策略成功地以不依赖血统的方式生成完整的FMDV基因组.
这种方法使用两种多重PCR反应,使其具有广泛的适用性.
这些原始组和方法提高了实验室对口腔疾病基因组特征的能力.

关键词:

全基因组口病一个小时纳米孔测序方式全基因组测序

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