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定制捕获测序面板适应牛津纳米孔MinION平台的自定义捕获测序面板
Vishal Kapoor1,2, Santiago Sanchez-Vicente3, William Donovan3
1Center for Infection and Immunity, Mailman School of Public Health, Columbia University, New York, USA. vk2040@cumc.columbia.edu.
Molecular biology reports
|March 3, 2026
概括
在便携式MiniON MK1C设备上,捕获序列增强了下一代序列测序 (NGS) 的灵敏度. 这种方法改善了用于临床诊断的病原体检测,提供与已建立的平台相匹配的性能.
科学领域:
- 临床微生物学 临床微生物学
- 基因组学就是基因组学.
- 分子诊断学 分子诊断学
背景情况:
- 下一代测序 (NGS) 提供广泛的病原体检测,但在临床使用中面临局限性,包括比PCR和复杂数据分析更低的灵敏度.
- 捕获测序丰富了向的核酸,与标准的元基因组NGS (mNGS) 相比,显著增加了灵敏度 (高达10,000倍).
- 这项研究探讨了将捕获测序应用到便携式牛津纳米孔技术MINION MK1C测序器.
研究的目的:
- 在MinION MK1C平台上评估捕获测序试验 (VirCapSeq-VERT和TBDCapSeq) 的性能.
- 为了比较灵敏度,病原体基因组覆盖率和MiniON MK1C捕获测序的读取输出与Illumina平台.
主要方法:
- 修改了基于Illumina的捕获测序协议的混合工作流,使其与ONT MinION测序适配器兼容.
- 使用人工和临床样本测试了两个捕获试验,VirCapSeq-VERT (脊椎动物病毒) 和TBDCapSeq (传播的病原体).
- 与Illumina NextSeq平台进行测试性能比较,包括灵敏度值和测序指标 (基因组覆盖范围,读数).
主要成果:
- 与mNGS相比,捕获丰富显著改善了使用MinION NGS的病原体检测.
- 在病原体检测方面,MinION MK1C表现出与Illumina NextSeq相等的灵敏度.
- 在MinION MK1C上,较长的读取长度弥补了较低的读取数量,从而获得了可比的病原体基因组覆盖范围.
结论:
- VirCapSeq和TBDCapSeq捕获试验在不同的测序平台上是有效的,包括MinION.
- 当与捕获测序技术相结合时,MinION平台显示了广泛的临床诊断潜力.
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