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Updated: Sep 8, 2025

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

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配对加减测序是一种超高吞吐量和精确的双链DNA分子测序方法

Alexandre Pellan Cheng^1,2,3, Itai Rusinek⁴, Aaron Sossin³

¹École de Technologie Supérieure, Montréal, Québec, Canada.

bioRxiv : the preprint server for biology

|August 20, 2025

概括

配对加减测序 (ppmSeq) 是一种具有成本效益的方法,可以准确地检测DNA中的低频单核酸变体 (SNV). 这项技术改善了在临床应用中增强基因组测序的双重恢复.

科学领域:

基因组学
分子生物学
生物信息学

背景情况:

区分真实生物变异 (SNV) 和测序错误至关重要,特别是在癌症检测和体质马赛克的低频变异中.
由于双重分子捕获率低,目前的双重测序方法需要大量的超级测序.

研究的目的:

引入配对加减测序 (ppmSeq) 作为对现有双重测序技术的进步.
评估ppmSeq在临床环境中检测低频变异的效率,错误率和适用性.

主要方法:

配对加减测序 (ppmSeq) 分区,并通过乳液PCR从单个分子中克隆地放大两个DNA链.
这两条线程都为单一的测序读取提供了帮助,从而使双重产量的线性扩展具有覆盖范围.
与现有的双重测序技术进行比较,并使用基因组和无细胞DNA评估错误率.

主要成果:

与领先的双重技术 (~5-11%) 相比,ppmSeq实现了更高的双重回收率 (44%±5.5%).
遗留SNV检测错误率在基因组DNA中低至7.98×10−8,而在无细胞DNA中低至3.5×10−7.
在癌症监测中证明有效的瘤信息和瘤原始循环瘤DNA (ctDNA) 检测.

结论:

ppmSeq提供了一个可扩展,成本高效,高可靠的错误纠正全基因组测序方法.
该技术可用于敏感的ctDNA检测,用于疾病监测和癌症特异性突变特征的识别.
ppmSeq非常适合具有挑战性的临床应用和需要高精度突变识别的人体遗传学.

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