目的:探讨第二代宏基因组下一代测序(mNGS)在肺部感染患者病原菌检测中的应用价值。
方法:对我院及上海市第五人民医院2021年1月至2023年5月收治的65例肺部感染病例进行回顾性分析。所有受试者都接受了mNGS,靶向下一代测序(tNGS),和常规微生物培养。进行了比较分析,以评估通过这些方法鉴定的病原体的多样性和数量,并评估其在肺部感染诊断中的诊断能力。
结果:mNGS在65例中的60例中成功确定了病因,与TNGS相比,在42例中产生了积极的结果,和传统的实验室培养,在24例中检测到病原体。在细菌属水平,mNGS辨别9属,11种,和92株病原菌,而tNGS确定了8个属,8种,和71个分离株。常规方法不太敏感,只检测6属,7种,33个分离株在真菌检测方面,mNGS鉴定出4种真菌,tNGS检测到4个念珠菌属的分离株,和常规方法鉴定了2个相同属的分离株。物种水平的病毒检测揭示了10个物种和46个mNGS分离株,而tNGS仅检测到3个物种和7个分离株。对于mNGS,以95%置信区间诊断肺部感染的受试者工作特征曲线(AUC)下面积为0.818(0.671至0.966),对于tNGS,0.668(0.475至0.860),常规培养为0.721(0.545至0.897)。mNGS在呼吸道感染的危重患者中显示出优越的诊断效能和病原体检测广度,通过减少诊断时间提供了显着的优势。mNGS的增强的敏感性和全面的病原体分析强调了其作为临床微生物学中领先的诊断工具的潜力。
OBJECTIVE: To explore the application value of the second-generation metagenomic next-generation sequencing (mNGS) in the
detection of pathogens in patients with pulmonary infection.
METHODS: We conducted a retrospective analysis of 65 pulmonary infection cases treated at our institution and the Fifth People\'s Hospital of Shanghai between January 2021 and May 2023. All subjects were subjected to mNGS, targeted next-generation sequencing (tNGS), and conventional microbiological culture. A comparative analysis was performed to evaluate the diversity and quantity of pathogens identified by these methodologies and to appraise their respective diagnostic capabilities in pulmonary infection diagnostics.
RESULTS: The mNGS successfully identified etiological agents in 60 of the 65 cases, compared to tNGS, which yielded positive results in 42 cases, and conventional laboratory cultures, which detected pathogens in 24 cases. At the bacterial genus level, mNGS discerned 9 genera, 11 species, and 92 isolates of pathogenic bacteria, whereas tNGS identified 8 genera, 8 species, and 71 isolates. Conventional methods were less sensitive, detecting only 6 genera, 7 species, and 33 isolates. In terms of fungal
detection, mNGS identified 4 fungal species, tNGS detected 4 isolates of the Candida genus, and conventional methods identified 2 isolates of the same genus. Viral
detection at the species level revealed 10 species and 46 isolates by mNGS, whereas tNGS detected only 3 species and 7 isolates. The area under the receiver operating characteristic curve (AUC) with 95% confidence intervals for diagnosing pulmonary infections was 0.818 (0.671 to 0.966) for mNGS, 0.668 (0.475 to 0.860) for tNGS, and 0.721 (0.545 to 0.897) for conventional culture.The mNGS demonstrates superior diagnostic efficacy and pathogen
detection breadth in critically ill patients with respiratory infections, offering a significant advantage by reducing the time to diagnosis. The enhanced sensitivity and comprehensive pathogen profiling of mNGS underscore its potential as a leading diagnostic tool in clinical microbiology.