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基于DSN的宿主DNA枯竭用于在下一代测序中同时检测DNA和RNA病原体

Han Ren^1,2, Xinlin Jiao², Ruizhi Wang³

¹Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.

Letters in applied microbiology

|January 10, 2026

概括

一种新的双重特异核酶 (DSN) 方法在临床下一代测序 (NGS) 中显著丰富了微生物读数. 这种方法通过减少宿主核酸污染在元基因组和向测序中提高了病原体检测灵敏度.

科学领域:

分子生物学分子生物学
基因组学就是基因组学.
临床诊断临床诊断临床诊断

背景情况:

核酸测序对于临床病原体检测至关重要.
样本中的高宿主DNA含量 (90-99%) 在元基因下一代测序 (mNGS) 中限制了微生物识别灵敏度.

研究的目的:

开发和验证一种基于双重特异核酶 (DSN) 的新方法,用于减少临床测序中的宿主核酸含量.
为了提高元基因性NGS (mNGS) 和向性NGS (tNGS) 的病原体检测灵敏度.

主要方法:

开发了一种双重特定核酶 (DSN) 工作流程,以选择性地降解宿主DNA.
优化包括测试回火温度,驱动DNA (Cot-1 DNA和rProbe) 和四甲基化物 (TMAC) 度.
该方法使用模拟微生物群落和临床样本进行了验证.

主要成果:

在mNGS中,DSN方法增加了病原体每百万读数 (RPM) 的2至3倍.
在基于探针的向NGS (tNGS) 中观察到病原体RPM的3到10倍的更实质性的改善.
最佳条件包括65°C的火,Cot-1 DNA和rProbe的特定度,以及150 mmol·L-1 TMAC.

结论:

在临床NGS中,DSN工作流提供了一种快速 (<30分钟) 和低损失的微生物丰富方法.
这种技术通过减少宿主DNA干扰,显著提高了病原体检测灵敏度.
通过将额外的人类基因组重复区域纳入驱动DNA,可以进一步提高宿主耗尽效率.

关键词:

双重体特异性核酶中的一个.主体消耗主体消耗 mNGSS 在线观看病原体检测检测病原体的检测 tNGS是什么意思 tNGS是什么意思

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