AECOPD患者医院感染病原学及微生物组学

Distribution of pathogens and microbiome of nosocomial infection in patients with AECOPD

目的比较在慢性阻塞性肺疾病急性加重期(AECOPD)病原体诊断层面不同检验试验方法,从而探讨细菌感染在AECOPD中的作用及发病机制。方法回顾性分析青海大学附属医院2016年1月-2019年9月2345例住院患者慢性阻塞性肺疾病(COPD)合并肺部感染的临床资料,分析其病原菌分布特征;选取2019年住院患者6例诊断为AECOPD的患者,采集肺泡灌洗液进行细菌培养,同时,对肺泡灌洗液进行微生物多样性高通量二代测序,从而比较微生物种类和分布情况。结果研究对象共纳入1592例AECOPD患者的病历,整理分析感染病原体种类,分别为革兰阴性菌包括肺炎克雷伯菌、鲍氏不动杆菌、铜绿假单胞菌等,革兰阳性菌包括肺炎链球菌,金黄色葡萄球菌;临床采集的6例AECOPD患者的肺泡灌洗液标本进行培养和测序,通过肺泡灌洗液标本培养检出的病原体分别为鲍氏不动杆菌和肺炎克雷伯菌,通过测序技术,细菌分类门水平的主要为蛋白质细菌、厚壁菌、放线菌门、拟杆菌、栖热菌门、梭杆菌门、蓝细菌、绿弯菌门、浮霉菌门、酸杆菌;属层面主要的病原体为链球菌、棒状杆菌、克雷伯菌、乳酸菌、贪铜菌属、假丝酵母、苍白杆菌属、嗜血杆菌、普雷沃菌属、葡萄球菌、通过聚类分析后热图结果显示嗜血杆菌、链球菌、假丝酵母、苍白杆菌属等在不同的样本中序列较高。结论从纵向患者人群数据,以及横向实验室数据进行比较,探讨在AECOPD病原体诊断层面,测序技术可能更优越于传统培养方法。

OBJECTIVE To compare the efficacy of different methods in the diagnosis of AECOPD and to explore the role and pathogenesis of bacterial infection in patients with AECOPD.METHODS The clinical data of 2345 COPD patients complicated with pulmonary infection in a general hospital in Qinghai Province from 2016 to 2019 was retrospectively analyzed to explore the distribution characteristics of pathogenic bacteria.Bronchoalveolar lavage fluid(BALF)from six hospitalized patients diagnosed as AECOPD in 2019 were collected and cultured.BALF samples were also sequenced by high-throughput second-generation sequencing at the same time.RESULTS A total of 1592 cases of COPD patients with pulmonary infection were included in the study.The types of pathogens included Gram negative bacteria such as Klebsiella pneumoniae,Acinetobacter baumannii and Pseudomonas aeruginosa,and Gram-positive bacteria containing Streptococcus pneumoniae and Staphylococcus aureus.A.baumannii and K.pneumoniae were detected from the BALF samples from 6 patients with AECOPD by bacterial cultivation.The Phyla of the pathogens detected by second-generation sequencing technology were Protein bacteria,Firmicum,Actinobacteria,Bacteroides,Thermophilia,Fusobacteria,Cyanobacteria,Curvularia,Fusobacteria,and Acidobacteria;the main pathogens at genus level were Streptococcus,Corynebacterium,Klebsiella,Lactic acid bacteria,Cuprobacter,Candida,Pallidobacterium,Haemophilus,and Pleurotus.The heat map of community composition at genus level showed that the abundance of Haemophilus,Streptococcus and Candida and Pallidobacterium was high in all patients.CONCLUSION From the longitudinal patient population data,as well as the horizontal laboratory data comparison,sequencing technology may be superior to the traditional culture method in the detection of pathogens in patients with AECOPD.