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在微生物无细胞DNA的元基因组测序的矩阵内图书馆准备

Paul L Babb¹, Jamilla Akhund-Zade¹, Damek Spacek¹

¹Karius, Inc., Redwood City, California, USA.

Journal of clinical microbiology

|November 28, 2025

概括

卡里乌斯-4化学提供了一种更快,更清洁,更敏感的方法来测序微生物无细胞DNA (mcfDNA). 这种无DNA提取的工作流显著提高了mcfDNA产量,减少了污染,降低了传染病诊断和生物标志物发现的成本.

科学领域:

微生物学微生物学
基因组学就是基因组学.
生物信息学是一种生物信息学.

背景情况:

微生物无细胞DNA (mcfDNA) 的甲基因组测序有助于识别病原体和抗菌素耐药性.
目前的方法由于复杂的工作流程,来自宿主DNA的高成本和污染而面临限制.
这些挑战阻碍了mcfDNA测序的可扩展部署.

研究的目的:

开发和评估Karius Helion-4化学 (Helion-4),用于mcfDNA测序的矩阵内工作流程.
克服现有的元基因组测序工作流程的局限性.
为了实现高效和可扩展的mcfDNA在传染病,微生物组分析和生物标志物发现中的应用.

主要方法:

开发了Helion-4,一种没有DNA提取的样本对DNA测序库工作流.
将Helion-4与两个基于提取的工作流程和使用36个等离子体样本的之前的Karius平台进行了比较.
评估工作流效率,DNA产量,污染水平和读取丰富度.

主要成果:

-4在5.25-6.1小时内实现了对96个样本的端到端图书馆构建,使用最少的实践时间.
与其他方法相比,恢复了58至817倍的内源性mcfDNA,并证明外源性DNA污染率为1.8至6倍.
丰富了mcfDNA读取的部分60至164倍,大大降低了测序成本.

结论:

-4为mcfDNA测序提供了一种简单,高效和可扩展的方法.
该技术克服了主要的障碍,使小量标本的例行处理成为可能.
在各种医疗保健环境中促进广泛的,高分辨率的实时微生物分析.

关键词:

这是一个DNA库协议,一个DNA图书馆协议.没有提取的自由提取.在矩阵中的矩阵.液体活检活检液体活检转基因组学是指转基因组学.微生物无细胞DNA的DNA.

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