Yang Jing, Wang Yan, Yang Li, Wu Jun
Beijing Jishuitan Hospital, Capital Medical University, Beijing, China.
Department of Laboratory Medicine, Beijing Jishuitan Hospital, Peking University Fourth School of Clinical Medicine, Beijing, China.
Microbiol Spectr. 2025 Jul;13(7):e0175124. doi: 10.1128/spectrum.01751-24. Epub 2025 May 21.
Lower respiratory tract infection (LRTI) is a serious global public health issue and poses significant challenges for detection and diagnosis. We validated the detection performance of targeted next-generation sequencing (tNGS) based on multiplex PCR using simulated microbial sample panels and clinical samples, providing a theoretical basis for promoting and applying tNGS in clinical diagnosis. We used a series of simulated microbial sample panels to validate the analytical validity of tNGS comprehensively. We also use tNGS to test respiratory specimens from 108 diagnosed or suspected LRTI patients to validate its clinical validity in diagnosing LRTI. Finally, we summarize the drug-resistance genes obtained from tNGS, the detection cost, and the turnaround time. tNGS has good analytical specificity, sensitivity, and precision. It has good stability when stored under low-temperature conditions. Using the composite diagnostic criteria as the gold standard, our internal tNGS platform has a sensitivity of 84.38%, specificity of 91.67%, positive predictive value of 98.78%, and negative predictive value of 42.31%. In terms of turnaround time, tNGS (about 16 h) and metagenomic next-generation sequencing (about 24 h) are similar, both significantly superior to traditional microbial detection methods (3-5 days). The cost of tNGS is approximately one-fourth of metagenomic next-generation sequencing. As a novel method with acceptable performance and cost, tNGS can compensate for the shortcomings of commonly used pathogen detection schemes in clinical practice, and its application prospects are worth looking forward to.
Lower respiratory tract infection (LRTI) is a serious global public health problem, and detecting its pathogenic microorganisms is difficult. Targeted next-generation sequencing (tNGS) is a rising star in microbial detection, with enormous potential. To understand the detection performance of tNGS and provide a theoretical basis for promoting its application in clinical diagnosis, this study prepared simulated microbial sample panels using reference materials to evaluate the analytical and clinical validity of tNGS. Our research suggests that tNGS has good analytical specificity and sensitivity, precision, and stability. Additionally, it can reliably detect common LRTI pathogens. It has advantages in identifying co-infections and atypical pathogens. Moreover, tNGS significantly reduces turnaround time, allowing faster treatment. In summary, tNGS is expected to be used in clinical practice to diagnose and manage LRTI.
下呼吸道感染（LRTI）是一个严重的全球公共卫生问题，在检测和诊断方面带来了重大挑战。我们使用模拟微生物样本板和临床样本验证了基于多重PCR的靶向新一代测序（tNGS）的检测性能，为tNGS在临床诊断中的推广和应用提供了理论依据。我们使用一系列模拟微生物样本板全面验证了tNGS的分析有效性。我们还使用tNGS检测了108例已确诊或疑似LRTI患者的呼吸道标本，以验证其在诊断LRTI方面的临床有效性。最后，我们总结了从tNGS获得的耐药基因、检测成本和周转时间。tNGS具有良好的分析特异性、灵敏度和精密度。在低温条件下储存时具有良好的稳定性。以综合诊断标准作为金标准，我们的内部tNGS平台灵敏度为84.38%，特异性为91.67%，阳性预测值为98.78%，阴性预测值为42.31%。在周转时间方面，tNGS（约16小时）和宏基因组新一代测序（约24小时）相似，两者均显著优于传统微生物检测方法（3 - 5天）。tNGS的成本约为宏基因组新一代测序的四分之一。作为一种性能和成本均可接受的新方法，tNGS可以弥补临床实践中常用病原体检测方案的不足，其应用前景值得期待。
下呼吸道感染（LRTI）是一个严重的全球公共卫生问题，检测其致病微生物具有困难。靶向新一代测序（tNGS）是微生物检测领域的一颗新星，具有巨大潜力。为了解tNGS的检测性能并为其在临床诊断中的应用提供理论依据，本研究使用参考材料制备模拟微生物样本板，以评估tNGS的分析和临床有效性。我们的研究表明，tNGS具有良好的分析特异性、灵敏度、精密度和稳定性。此外，它能够可靠地检测常见的LRTI病原体。在识别合并感染和非典型病原体方面具有优势。而且，tNGS显著缩短了周转时间，能够更快地进行治疗。总之，tNGS有望用于临床实践中诊断和管理LRTI。
