开发和验证基于Amplicon的呼吸道同胞病毒基因组测序协议
Guglielmo Ferrari1, Great Romano1, Antonino Maria Guglielmo Pitrolo1
1Microbiology and Virology Department, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
Journal of medical virology
|August 20, 2025
概括
我们从临床样本中直接开发了一种全基因组测序的新方法. 这种改进的方法增强了对这种常见的儿童呼吸道感染的RSV监测和分子流行病学工作.
科学领域:
- 病毒学
- 基因组学
- 流行病学
背景情况:
- 人类呼吸道同胞病毒 (RSV) 是儿童严重呼吸道感染的主要原因.
- 下一代测序 (NGS) 技术对于病原体监测和监测越来越重要.
- 全基因组测序 (WGS) 是跟踪RSV演变的重要工具.
研究的目的:
- 报告一个改进的,高通量RSV全基因组测序 (WGS) 试验.
- 使用短读测序平台直接对临床样本进行测试的验证.
- 为分子流行病学和RSV监测提供强有力的协议.
主要方法:
- 开发一种高通量RSV-WGS测定方法,扩大覆盖整个基因组的三个不同的安普利康.
- 在2022年11月至2024年3月期间采集的100个RSV阳性临床样本的验证.
- 用低病毒载量样本评估检测性能 (量化值周期为27-32).
主要成果:
- 即使病毒载量低,测试也显示出高可复制性 (约95%).
- 具有意义的测序结果获得了高中位覆盖深度 (> 12000 ×) 和映射读数 (> 1 × 10 ^ 6).
- 测序的基因组在整个基因组中至少达到98%的覆盖率.
结论:
- 开发的RSV-WGS试验是有效的,适用于短读测序平台.
- 该方案易于执行,对于RSV的分子流行病学和监测具有高度可靠性.
- 这种方法有助于更好地追踪RSV的演变和传播模式.
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