在MinION平台上使用两种不同的化学物质对Orthoavulavirus javaense直接RNA测序的比较
Megan C Mears1, Quentin D Read2, Abhijeet Bakre1
1Exotic and Emerging Avian Viral Disease Research Unit, Southeast Poultry Research Laboratories, US National Poultry Research Center, 934 College Station Road, Athens, GA 30605, United States.
Journal of virological methods
|December 26, 2024
概括
使用牛津纳米孔技术 (ONT) 的直接RNA测序 (DRS) MinION成功地测序了禽类病原体Orthoavulavirus javaense (OAVJ) 的完整病毒基因组. 这种进步为快速的现场诊断和表皮转录组研究提供了潜力.
科学领域:
- 病毒学 病毒学
- 基因组学就是基因组学.
- 分子生物学分子生物学
背景情况:
- 由于经济原因,在家禽中快速识别病毒病原体至关重要,尤其是在疫情爆发期间.
- 目前的下一代测序方法是实验室密集的,不适合现场应用.
- 牙买加的orthoavulavirus javaense (OAVJ) 导致纽卡斯尔病,这是全世界家禽的重大威胁.
研究的目的:
- 研究使用牛津纳米孔技术 (ONT) MinION设备进行直接RNA测序 (DRS) 的可行性,用于OAVJ的全长病毒RNA基因组测序.
- 评估新的ONT测序化学和基调用工作流程用于病毒RNA分析的性能.
主要方法:
- 直接RNA测序 (DRS) 在OAVJ上使用ONT MinION设备进行.
- 一个定制的OAVJ专用适配器被开发出来,并与ONT DRS套件一起使用.
- 采用了新的ONT SQK-RNA004化学和流动细胞,以及改进的基础调用工作流.
主要成果:
- 该研究成功地证明了OAVJ病毒RNAs的捕获和测序,使用DRS与自定义适配器和ONT套件.
- 与以前的方法相比,新的ONT SQK-RNA004化学和基调用工作流显示出更好的读取质量和长度.
- 这标志着使用DRS.的Paramyxoviridae家族成员几乎全长的病毒RNA基因组测序方法的首次报告.
结论:
- 使用ONT MinION技术的DRS是一种有前途的方法,用于对OAVJ的完整病毒RNA基因组进行测序.
- 虽然需要进一步改进以实现广泛的现场适应,但DRS具有快速禽类病原体诊断的潜力.
- 这种技术为未来对OAVJ表体转录组及其在疾病发病过程中的作用的研究开辟了道路.
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