二代测序技术在社区获得性肺炎病原学诊断中的应用价值

Application value of next-generation sequencing technology in etiological diagnosis of community-acquired pneumonia

目的研究二代测序(next-generation sequencing,NGS)技术在社区获得性肺炎(community-acquired pneumonia,CAP)病原学诊断中的应用价值。方法采用回顾性研究方法,选取武汉市中心医院呼吸与危重症医学科2020年12月至2021年8月采用支气管镜检查收集肺泡灌洗液标本并行NGS送检的64例社CAP患者作为研究对象,对其临床相关资料进行分析,比较NGS技术与其他传统病原学检测手段在诊断CAP患者病原体中的差异。结果64例肺泡灌洗液NGS送检患者中,阳性55例、阴性9例。NGS有41例患者检出细菌27种,传统细菌培养有17例患者检出细菌6种,NGS细菌检出率显著高于传统细菌培养(64.1%vs.26.6%,P<0.001)。NGS有27例患者检出真菌,传统真菌培养31例检出真菌,NGS真菌检出率与传统真菌培养之间无显著差异(42.2%vs.48.4%,P=0.48)。NGS检出病毒29例,嗜肺军团菌1例,鹦鹉热衣原体2例,结核分枝杆菌6例,未检出支原体及衣原体;传统病原学检测检出病毒2例,结核分枝杆菌4例,肺炎衣原体2例,未检出鹦鹉热衣原体、嗜肺军团菌、肺炎支原体。结论NGS可以同时检测细菌、真菌及病毒等非典型病原体,相较于传统病原学检测方法,其检测病原微生物范围广、种类丰富,对CAP患者的病原学诊断具有重要意义,但检出病原体仍需结合临床分析以确定其是否为致病菌。

Objective To investigate the application value of next-generation sequencing(NGS)technology in the etiological diagnosis of community-acquired pneumonia(CAP).Methods A retrospective study was conducted to analyze the clinical data of 64 patients with CAP whose alveolar lavage fluid samples were collected by bronchoscopy from Department of Respiratory and Critical Care Medicine,Wuhan Central Hospital from December 2020 to August 2021 for NGS examination,and to compare the differences between NGS technology and other traditional etiological detection methods in the diagnosis of pathogens in patients with CAP.Results Among 64 patients with alveolar lavage fluid NGS,55 cases were positive and 9 cases were negative.In NGS,27 bacterial species were detected in 41 patients,and 6 bacteria were detected in 17 patients with traditional bacterial culture,and the detection rate of NGS bacteria was significantly higher than that of unified bacterial culture(64.1%vs.26.6%,P<0.001).There were 27 patients with NGS and 31 cases with traditional fungal culture,and there was no significant difference between the detection rates of NGS fungi and traditional fungal culture(48.4%vs.42.2%,P=0.48).NGS detected virus in 29 cases,Legionella pneumophila in 1 case,chlamydia pti in 2 cases,Mycobacterium tuberculosis in 6 cases,and mycoplasma and chlamydia were not detected.Traditional etiological tests detected virus in 2 cases,Mycobacterium tuberculosis in 4 cases,chlamydia pneumonia in 2 cases,Chlamydia pti,Legionella pneumophila,and mycoplasma pneumoniae were not detected.Conclusion NGS can be used for the detection of atypical pathogens such as bacteria,fungi and viruses at the same time.Compared with traditional pathogen detection methods,NGS can detect a wide range of pathogenic microorganisms with rich species,which is of great significance in the etiological diagnosis of patients with community-acquired pneumonia.However,pathogens detected by NGS need to be combined with clinical analysis to determine whether they are pathogenic bacteria.