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基于16S rRNA测序技术的呼吸机相关性肺炎患者粪便与肺泡灌洗液菌群相关性分析

[Analysis of correlation between fecal and alveolar lavage fluid flora of ventilator-associated pneumonia patients based on 16S rRNA sequencing technology].

作者信息

Lei Mengmeng, Zhang Xiaoya, Yang Xiaojuan, Jing Pei, Yang Mingyue, Yang Xiaojun

机构信息

School of Clinical Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China.

Department of Critical Care Medicine, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China. Corresponding author: Yang Xiaojun, Email:

出版信息

Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2021 Feb;33(2):169-173. doi: 10.3760/cma.j.cn121430-20201010-00662.

Abstract

OBJECTIVE

To investigate the changes and correlation of intestinal and pulmonary microecological structures in patients with ventilator-associated pneumonia (VAP).

METHODS

A prospective observational study was conducted. Thirty-one patients with VAP admitted to the department of critical care medicine of General Hospital of Ningxia Medical University from May 1st 2019 to May 1st 2020 were enrolled. Feces and alveolar lavage fluid samples from patients with the same day, feces and alveolar lavage specimen flora composition and the structure of biological information analysis by 16S rRNA sequencing technologies, the comprehensive sequencing results, and clinical data of patients were analyzed.

RESULTS

(1) The diversity (abundance and diversity) of flora in the alveolar lavage fluid of VAP patients was higher than that of fecal flora. Among them, Ace index, Chao index and Shannon index describing the abundance of flora showed statistically significant differences [Ace index: 305.89 (214.39, 458.66) vs. 204.51 (165.15, 247.61), Chao index: 259.83 (194.20, 459.31) vs. 187.67 (153.28, 234.01), Shannon index: 3.01 (2.39, 3.54) vs. 2.55 (1.86, 2.95), all P < 0.05], but there was no significant difference in Simpson index describing diversity [0.14 (0.08, 0.27) vs. 0.19 (0.10, 0.33), P > 0.05]. (2) In the sequencing results of feces and alveolar lavage fluid of VAP patients, there were some intestinal related bacteria groups with high abundance, such as Escherichia-Shigella, Faecalibacterium, Bacteroides, and Lachnospira, etc. (3) In 31 VAP patients, suspicious pathogenic bacteria was found in 20 cases (6 cases of Streptococcus viridans, 5 cases of Escherichia coli, 3 cases of Klebsiella pneumoniae, 3 cases of Acinetobacter baumannii, 2 cases of Staphylococcus aureus, 1 case of Pseudomonas aeruginosa), and the same suspected pathogens also existed in the 17 patients' alveolar lavage and waste sequencing. (4) Fourteen VAP patients combined with sepsis, 14 patients without sepsis were selected for sample size matching. The results showed that, Jaccard similarity index to describe lung-correlation of intestinal flora in VAP with sepsis group was significantly elevated, and the difference was statistically significant (0.24±0.08 vs. 0.19±0.06, P < 0.01).

CONCLUSIONS

There is a certain correlation between pulmonary and intestinal flora in VAP patients. In addition to the exclusion of pulmonary infection caused by environmental and upper respiratory micro-inhalation, the lower digestive tract may also be source of infection.

摘要

目的

探讨呼吸机相关性肺炎(VAP)患者肠道与肺部微生态结构的变化及相关性。

方法

进行一项前瞻性观察性研究。选取2019年5月1日至2020年5月1日在宁夏医科大学总医院重症医学科住院的31例VAP患者。采用16S rRNA测序技术对患者同日粪便及肺泡灌洗液样本、粪便及肺泡灌洗标本菌群组成及生物信息结构进行分析,并对综合测序结果与患者临床资料进行分析。

结果

(1)VAP患者肺泡灌洗液中菌群的多样性(丰富度和多样性)高于粪便菌群。其中,描述菌群丰富度的Ace指数、Chao指数和Shannon指数差异有统计学意义[Ace指数:305.89(214.39,458.66)对204.51(165.15,247.61),Chao指数:259.83(194.20,459.31)对187.67(153.28,234.01),Shannon指数:3.01(2.39,3.54)对2.55(1.86,2.95),均P<0.05],但描述多样性的Simpson指数差异无统计学意义[0.14(0.08,0.27)对0.19(0.10,0.33),P>0.05]。(2)VAP患者粪便和肺泡灌洗液测序结果中,存在一些高丰度的肠道相关菌群,如埃希-志贺菌属、粪杆菌属、拟杆菌属、毛螺菌属等。(3)31例VAP患者中,20例检出可疑病原菌(草绿色链球菌6例、大肠埃希菌5例、肺炎克雷伯菌3例、鲍曼不动杆菌3例、金黄色葡萄球菌2例、铜绿假单胞菌1例),17例患者肺泡灌洗及粪便测序中也存在相同的可疑病原菌。(4)选取14例合并脓毒症的VAP患者,14例未合并脓毒症患者进行样本量匹配。结果显示,描述VAP合并脓毒症组肠道菌群与肺部相关性的Jaccard相似性指数显著升高,差异有统计学意义(0.24±0.08对0.19±0.06,P<0.01)。

结论

VAP患者肺部与肠道菌群之间存在一定相关性。除排除环境及上呼吸道微吸入所致肺部感染外,下消化道也可能是感染源。

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