针对包括SARS-CoV-2在内的肉食病原体的MinION适应的tNGS面板
Nelly O Elshafie1, Jobin J Kattoor2, Janetta Kelly3
1Willie M. Reed Animal Disease Diagnostic Laboratory, Department of Comparative Pathobiology, College of Veterinary Medicine, Purdue University, West Lafayette, IN 47907, USA.
Pathogens (Basel, Switzerland)
|January 28, 2026
概括
野生动物病原体监测通过在牛津纳米孔MiniON平台上的新的,负担得起的下一代定向测序 (tNGS) 方法得到了增强. 这种方法可以同时检测SARS-CoV-2并对动物进行更广泛的病原体查.
科学领域:
- 兽医医学 兽医医学 兽医医学
- 基因组学就是基因组学.
- 流行病学 流行病学
背景情况:
- 对像SARS-CoV-2这样的病原体进行有效的野生动物监测至关重要.
- 目前的方法,如核酸放大试验 (NAAT),在检测共感染或新型病原体方面存在局限性.
- 需要可负担得起和灵活的工具来全面检测野生动物病原体.
研究的目的:
- 为了适应一个针对性下一代测序 (tNGS) 面板的 mesocarnivores 牛津纳米孔技术 (ONT) MinION 平台.
- 将这个tNGS面板与SARS-CoV-2全基因组测序试验相结合,用于同时检测和变种识别.
- 评价野生动物病原体监测这一综合工作流程的性能.
主要方法:
- 将一个经过验证的肉食动物tNGS面板从Ion Torrent调整到ONT MinION平台上,优化了多重复合的条件.
- 在图书馆准备之前,与tNGS面板进行集成的SARS-CoV-2全基因组测序.
- 使用已知SARS-CoV-2稀释的人工样本评估分析灵敏度和与NAAT结果对比的诊断性能.
主要成果:
- 在MinION tNGS工作流中,检测到SARS-CoV-2到Ct 34,并实现了Ct ≤24的≥99%基因组覆盖率.
- 与SARS-CoV-2同时检测其他病原体是可能的.
- 在测试的野生动物样本中,诊断灵敏度为96.7%,特异性为100%.
结论:
- 集成的MinION tNGS工作流提供了一种低成本,可扩展和高效的方法,用于全面监测野生动物病原体.
- 这种方法可以同时检测SARS-CoV-2,识别变种,并查其他病原体.
- 优化的工作流改善了交付时间,降低了成本,使其适合于低量监控工作.
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