目的:初步建立纳米孔16S测序技术检测下呼吸道感染细菌性病原的流程和方法，并评估其可行性。方法:收集北京医院呼吸与危重症医学科2019年7月至2020年9月就诊的33例下呼吸道感染患者的支气管肺泡灌洗液标本，进行纳米孔16S测序。采用χ2检验对16S测序结果和细菌培养结果进行统计分析，比较病原检出率以及病原种类，对纳米孔测序在病原体检测的应用进行评估。结果:33例患者的16S测序病原检出率高于传统的细菌培养［75.8%（25/33），45.5%（15/33），χ2=5.140，n P<0.05］。25份纳米孔16S测序阳性样本中，共检测出16种病原体，主要包括副流感嗜血杆菌、流感嗜血杆菌、肺炎链球菌、嗜麦芽窄食单胞菌、鲍曼不动杆菌、琼氏不动杆菌、金黄色葡萄球菌、肺炎克雷伯菌、粪肠球菌、鹑鸡肠球菌、纹带棒状杆菌、副胞内分枝杆菌、黏质沙雷菌、n Insuavis无色杆菌、默氏枸橼酸杆菌以及肺炎支原体，多于临床培养检测的6种病原体，包括副流感嗜血杆菌、鲍曼不动杆菌、铜绿假单胞菌、金黄色葡萄球菌、肺炎克雷伯菌和嗜麦芽窄食单胞菌（χ2=7.949，n P<0.05）。纳米孔16S测序比对至种水平的序列占属水平的80.0%（60.0%，86.0%），与细菌培养结果的一致率为33.3%（11/33）。n 结论:本研究建立的纳米孔16S扩增子测序流程能够从下呼吸道感染患者的支气管肺泡灌洗液中快速鉴定细菌性病原。纳米孔16S扩增子测序病原检出率高，检出病原种类多于细菌培养，且可以将多数细菌鉴定至种水平。该技术是一种非常有潜力的平台，具有广阔的应用前景。“,”Objective:To establish the method for detecting lower respiratory infections (LRIs) bacterialpathogens using nanopore sequencing, and evaluate the feasibility of this method.Methods:Bronchoalveolar lavage fluid (BALF) samples from 33 patients with LRIs who visited the Department of Respiratory and Critical Care Medicine of Beijing Hospital from July 2019 to September 2020 were collected.Nanopore 16S amplicon sequencing were performed on these samples. In order to evaluate the clinical value of the nanopore sequencing, χn 2 test was used to analyze the pathogen differences between the detection rate and pathogen types results found with using the nanopore 16S sequencing and the results found with bacterial culture.n Results:The process and method of nanopore sequencing used in the detection of the LRIs pathogens were established. The pathogen detection rate of the 16S sequencing was higher than that of the traditional bacterial culture (75.8% [25/33], 45.5% [15/33], χ2=5.140,n P<0.05). From the 25 positive samples found with nanopore 16S sequencing, 16 pathogens were detected, includingn Haemophilus parainfluenzae, n Haemophilus influenzae, n Streptococcus pneumoniae, n Streptomonas maltophilia, n Acinetobacter baumannii, and n Acinetobacter junii, n Staphylococcus aureus, n Klebsiella pneumoniae, n Enterococcus faecalis, n Enterococcus gallinarum, n Corynebacterium striatum, n Mycobacterium paraintracellulare, n Serratia marcescens, n Achromobacter insuavis, n Citrobacter murliniae and n Mycoplasma pneumoniae. More than 6 pathogens were tested in clinical culture, including n Haemophilus parainfluenzae, n Acinetobacter baumannii, n Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae and n Streptomonas maltophilia (χ2=7.949, n P<0.05). 16S sequencing aligned to species level sequences accounted for 80.0 (60.0, 86.0)% of the genus level. The results obtained by using16S sequencing and bacterial culture were consistent in 11 (33.3%) samples.n Conclusions:Nanopore 16S amplicon sequencing can quickly identify pathogenic bacteria from BALF in LRIs patients. Nanopore 16S amplicon sequencing has a high detection rate, it can detect more pathogens than traditional bacterial culture, and it can also identify most bacteria to the species level. This technology is a very promising platform with broad application prospects.