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新的目标下一代测序 (NGS) 面板用于检测急性呼吸道感染病原体
Maksim Koryukov1, Andrey Kechin1, Mariya Gordukova2
1Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia, 630090; Novosibirsk State University, Novosibirsk, Russia, 630090.
Acta tropica
|February 4, 2026
概括
一种新的下一代定向测序 (NGS) 试验可以检测32种病毒和13种导致呼吸道感染的细菌. 这种先进的分子诊断工具改善了病原体的识别,以改善患者护理和公共卫生监测.
科学领域:
- 分子生物学分子生物学
- 传染性疾病 传染性疾病
- 基因组学就是基因组学.
背景情况:
- 呼吸道感染对全球健康造成重大负担,导致广泛的发病率和死亡率.
- 当前的诊断方法往往在检测各种呼吸道病原体的效率和范围上有局限性.
- 迫切需要改进的诊断工具来识别呼吸系统疾病的致病因子.
研究的目的:
- 开发和验证内部针对性下一代测序 (NGS) 试验,用于同时检测多种呼吸道病原体.
- 通过改进原料设计和PCR条件来优化试验的性能.
- 与传统的多重PCR面板相比,评估NGS试验的临床实用性.
主要方法:
- 使用NGS-PrimerPlex设计了117个原始对,针对32种病毒和13种细菌,分为三种多重PCR反应.
- 进行了in silico和in vitro评估,包括单体PCR和聚烯胺凝电泳,以评估原料的特异性和覆盖范围.
- 系统优化PCR变量 (primer/dNTP度,循环数,回火温度,速) 和反转录协议.
- 使用120名患者cDNA样本验证了测定,并将结果与商业多重PCR面板进行比较.
主要成果:
- 针对性的NGS测定在60%的阳性样本中显示出与PCR完全一致,当排除具有高Cq值的样本时,该比例上升到74%.
- 该试验通过商业PCR检测出13个阴性样本中的病原体,并在47个PCR阴性样本中检测出额外的病原体.
- 总的来说,NGS测定成功地在120个患者样本中的86个中确定了至少一个向病原体.
结论:
- 开发的向NGS测定提供了一种全面和有效的方法来检测广泛的呼吸道病原体.
- 这种分子诊断方法显示出增强呼吸道疾病监测和临床诊断的巨大潜力.
- 这些发现为未来开发针对传染病的基于NGS的先进诊断解决方案提供了坚实的基础.
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