开发和全面评估培养独立的,基于长安普利康的目标下一代测序方法,用于预测结核病中的抗菌素耐药性
Lulu Zhang1,2, Xia Yu3, Chi Zhang1,2
1NHC Key Laboratory of Systems Biology of Pathogens, National Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 102629, P. R. China.
Analytical chemistry
|December 21, 2024
概括
一种新的独立于培养的下一代向测序 (tNGS) 方法可以准确预测结核病患者的抗菌素耐药性 (AMR). 这种优化的牛津纳米孔测序方法提供了快速的结果和与现有诊断测试的高度一致性.
科学领域:
- 基因组学和分子诊断的研究.
- 传染病研究 传染病研究
- 抗微生物耐药性 (AMR) 研究研究
背景情况:
- 结核病 (TB) 的多种抗微生物耐药性 (AMR) 概况需要先进的检测方法.
- 现有的诊断方法可能在多种药物之间全面识别AMR方面存在局限性.
研究的目的:
- 开发和优化一种文化独立的,基于长安普利康的目标下一代测序 (tNGS) 方法.
- 为了预测Mycobacterium结核病综合体 (MTBC) 内的16种药物的AMR.
- 评估优化的tNGS方法的性能和临床实用性.
主要方法:
- 使用多重PCR来丰富MTBC的20个基因区域.
- 在牛津纳米孔技术 (ONT) 和Illumina平台上进行了测序.
- 开发了定制的生物信息管道用于数据分析和报告;使用Q20+化学和R10.4.1流细胞优化了ONT.
主要成果:
- 优化的ONT tNGS方法只需要15个高质量读取每个目标基因,以准确识别变异.
- 实现了4小时50分钟的快速周转时间.
- 与表型药物敏感性测试 (pDST) 98.35%的一致性和与Xpert MTB/RIF测试的一致性;有效地确定了对临床病原体干扰具有高耐药性的Mycobacterium物种.
结论:
- 优化的ONT tNGS方法为预测结核病中的AMR提供了快速,准确和全面的方法.
- 这种文化独立的方法简化了原料设计,并且与各种测序平台兼容.
- 在速度,准确性和AMR检测突变覆盖率方面,tNGS方法显著改进了以前的方法.
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