自动化可定制的图书馆准备下一代测序到一个开放的微流体平台的自动化
Anne Hoffmann1,2, Anke Timm2, Christopher Johnson3
1Institute of Interfacial Process Engineering and Plasma Technology, University of Stuttgart, Nobelstraße 12, 70569, Stuttgart, Germany.
Scientific reports
|July 26, 2024
概括
本研究引入了一种用于下一代测序 (NGS) 图书馆准备的自动化实验室芯片系统. 这种具有成本效益的解决方案通过有效分析无细胞DNA (cfDNA) 来提高癌症诊断.
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 医学诊断 医学诊断 医学诊断
背景情况:
- 下一代测序 (NGS) 对于癌症管理至关重要,特别是使用无细胞DNA (cfDNA) 监测治疗反应,因为它在检测罕见突变方面具有很高的灵敏度.
- 传统的NGS图书馆准备是复杂和耗时的,通常需要昂贵的,大规模的自动化不适合低到中等吞吐量实验室.
- 现有的自动化解决方案昂贵,需要专家处理,限制了小型实验室的可访问性.
研究的目的:
- 为NGS.提供一个自动化,低到中等吞吐量库准备工作流程的概念验证.
- 以商业上可用的,开放的lab-on-a-chip平台来展示一个替代的自动化解决方案.
- 在微流体环境中优化常见的图书馆准备步骤,用于cfDNA分析.
主要方法:
- 开发了一种基于芯片的实验室盒,集成可定制的PCR用于目标丰富,终端修复,适配器结合,基于磁珠的核酸净化和量化.
- 为微流体环境优化了标准库准备步骤.
- 通过使用已知突变的参考cfDNA在不同等位基因频率上验证了工作流.
主要成果:
- 自动化lab-on-a-chip工作流程成功执行了所有关键的库准备步骤.
- 自动化系统的Amplicon测序结果与手动处理具有很高的可比性 (Pearson r = 0.94).
- 该系统证明了与含有已知突变的cfDNA在不同等位基因频率的功能.
结论:
- 拟议的自动化lab-on-a-chip工作流提供了一个可行且具有成本效益的替代方案,用于NGS库准备在低到中等吞吐量设置.
- 这种微流体方法可以通过简化和自动化图书馆准备来促进NGS在常规癌症管理中的整合.
- 该技术有可能将NGS的应用范围扩大到癌症治疗监测临床诊断的研究之外.
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