相关实验视频
Updated: Jan 17, 2026

08:57
Amplicon Sequencing using the Long-Read Sequencing Technologies
Published on: August 29, 2025
495
使用长读测序技术进行幅测序
Morwasehla Modjadji1, Brendon Coenrad Mann1, Johannes Loubser1
1Department of Science and Innovation - National Research Foundation Centre of Excellence for Biomedical Tuberculosis Research, South African Medical Research Council Centre for Tuberculosis Research, Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University.
Journal of visualized experiments : JoVE
|September 15, 2025
概括
便携式测序为检测耐药结核病 (DR-TB) 提供了一种具有成本效益的方法. 这种方法可以准确识别高频耐药性突变,但可能会错过低频变异,影响资源有限的环境中的结核病诊断.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 传染病诊断 传染病诊断 传染病诊断
背景情况:
- 世界卫生组织 (WHO) 强调需要可访问的结核病 (TB) 诊断.
- 下一代测序 (NGS) 提高了耐药结核病 (DR-TB) 的检测,有针对性的NGS (tNGS) 专注于耐药性突变.
- 高成本的测序平台限制了低收入和中等收入国家的DR-TB诊断.
研究的目的:
- 用tNGS试验评估便携式测序对于DR-TB检测的实用性.
- 为了比较便携式 (长读) 测序与高精度 (短读) 测序的性能,用于识别结核病中与耐药性相关的变体.
主要方法:
- 从耐利芬素结核病 (RR-TB) 隔离物的DNA使用tNGS试验进行了放大.
- 放大产品在便携式测序平台和高精度短读平台上进行了测序.
- 生物信息学管道用于长读和短读数据被用于分析.
主要成果:
- 便携式测序方法成功识别了与高频电阻相关的变体.
- 便携式测序的结果与常见变体的参考短读测序平台的相关性很好.
- 在使用便携式测序方法检测低频电阻相关变异时观察到局限性.
结论:
- 便携式测序是DR-TB诊断的可行,具有成本效益的替代方案,特别是在资源有限的环境中.
- 虽然对高频变体有效,但需要进一步优化,以提高对低频变体在结核病耐药性检测中的灵敏度.
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