通过下一代测序检测到的插入,删除和删除-插入变体的确认
Lauren A Choate1, Alaa Koleilat1, Kimberley Harris1
1Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, United States.
Clinical chemistry
|August 11, 2023
概括
下一代测序 (NGS) 准确地检测到插入,删除和删除插入变体 (indels) 高达68bp. 对于NGS检测到的indels,桑格测序确认通常是不必要的,从而提高了效率.
科学领域:
- 基因组学就是基因组学.
- 分子生物学分子生物学
- 临床诊断 临床诊断 临床诊断
背景情况:
- 下一代测序 (NGS) 对于临床变种检测越来越准确.
- 然而,桑格测序仍然用于确认,增加了成本和延误.
- 对于插入,删除和删除-插入变体 (indels) 的NGS准确性存在有限的数据.
研究的目的:
- 与桑格测序相比,评估NGS用于indel检测的准确性和一致性.
- 在临床诊断环境中评估indels的NGS性能.
主要方法:
- 从NGS目标基因小组 (2015-2022) 进行了indel结果的回顾性分析.
- 对NGS和桑格测序进行临床和试验内验证的比较.
- 细分体大小,类型,变异频率,覆盖深度和基因组复杂性的分析.
主要成果:
- 在NGS和桑格测序之间100%的一致性超过490个indels (217个独特).
- 精确检测indels高达68bp,包括那些复杂的基因组区域.
- 在试验内验证过程中,indel检测的高可重复性 (100%).
结论:
- 在复杂的区域内,NGS可以准确地评估高达68bp的可报告价值,即使在复杂的地区.
- 在适当的NGS覆盖和等位基因频率值的情况下,对于indels的桑格测序确认通常不需要.
- 通过减少对确认桑格序列的需求,NGS可以简化临床测试工作流程.
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