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Updated: Jun 7, 2025

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开发基于标记的下一代测序临床试验,作为基于毛细管电泳测序的替代方案

Wei Cheng David Kuek¹, Chean Nee Chai¹, Wei Ming Jason Tham¹

¹Department of Laboratory Medicine, Molecular Diagnosis Centre, National University Health System, Singapore, Singapore.

Molecular genetics & genomic medicine

|November 19, 2024

概括

下一代测序 (NGS) 精确验证了89个样本中的88个样本,证明了其替代传统方法的实用性. 该iSeq-Nextera工作流显示多重复基因测试的承诺.

科学领域:

分子生物学分子生物学
基因组学就是基因组学.
生物技术是生物技术.

背景情况:

下一代测序 (NGS) 允许多重复用多个感兴趣区域 (ROI).
负担得起的NGS平台有助于探索从毛细管电泳 (CE) 和单基因测试到NGS的过渡.
该研究评估了Illumina iSeq 100在高通量样本验证方面的能力.

研究的目的:

评估迁移毛细管电泳 (CE) 和单基因测试对NGS的可行性.
为了验证Illumina iSeq 100用于多重样本分析的性能.
评估iSeq-Nextera的工作流程,以准确检测遗传变异.

主要方法:

从89个不同样本类型的档案样本中提取基因组DNA.
聚合酶连锁反应 (PCR) 放大使用内部原料.
使用Nextera XT套件进行图书馆准备,然后在Illumina iSeq 100上对磁珠进行清理和排序.

主要成果:

在89个样本中,88个样本实现了精确的测序和变异性基因鉴定.
一个样本的初始不一致性通过重新设计向异合体删除区域的原始程序来解决.
该工作流在样本验证和变种检测方面取得了很高的成功.

结论:

对于验证样品和识别遗传变异,iSeq-Nextera的工作流是准确的.
虽然对检测准确,但Nextera XT套件产生的变异性基因频率可能缺乏精度.
国家基因系统为以前由CE进行的多重基因分析提供了实用和高效的替代方案.

关键词:

这是Nextera XT.在iSeq 100 中,有100个字母.这是下一代测序.验证验证的时间

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