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Updated: Jan 18, 2026
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Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
Published on: August 29, 2014
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下一代测序改善了诊断 16S rRNA 基于片的微生物群分析临床样本与桑格测序相比
Huma Aftab1,2, Christian H Schouw2, Rimtas Dargis2
1Department of Clinical Microbiology, University Hospital Rigshospitalet, Copenhagen, Denmark.
APMIS : acta pathologica, microbiologica, et immunologica Scandinavica
|September 9, 2025
概括
牛津纳米孔技术 (ONT) 测序在临床样本中提供了改进的细菌病原体检测. 这种下一代测序方法与桑格测序相比,提高了单个和多个物种的识别能力.
科学领域:
- 微生物学 微生物学
- 基因组学就是基因组学.
- 临床诊断 临床诊断 临床诊断
背景情况:
- 16S核糖体RNA (rRNA) 基因测序对于识别细菌病原体至关重要.
- 桑格测序与多微生物样本作斗争,产生无法解释的结果.
- 下一代测序 (NGS) 为复杂样品提供了一个潜在的解决方案.
研究的目的:
- 评估牛津纳米孔技术 (ONT) 测序,用于在临床样本中检测细菌病原体.
- 将ONT测序的诊断性能与16SrRNA基因分析的桑格测序进行比较.
- 评估ONT在识别单菌和多微生物感染方面的能力.
主要方法:
- 101个临床样本在PCR放大16S rRNA基因后进行了桑格和ONT测序.
- 桑格序列使用BLAST与NCBI数据库进行了分析.
- 在使用EPI2ME Fastq 16S平台处理ONT数据.
主要成果:
- 与桑格测序 (59%) 相比,ONT测序显示出更高的阳性率 (72%).
- 在ONT和桑格测序之间的一致性为80%.
- ONT发现的多微生物样本 (13) 多于Sanger的 (5),包括Borrelia bissettiiae的一个病例,Sanger错过了.
结论:
- 在临床环境中,ONT测序改善了细菌病原体的检测,包括多微生物感染.
- 与传统的桑格测序相比,ONT为细菌识别提供了更全面的诊断工具.
- 这项研究支持采用ONT测序作为一种常规的诊断方法,用于增强病原体检测.
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