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中国防痨杂志 ›› 2021, Vol. 43 ›› Issue (4): 357-363.doi: 10.3969/j.issn.1000-6621.2021.04.011

• 论著 • 上一篇    下一篇

结核分枝杆菌感染对痰液菌群结构的影响

张晨晨, 谭卫国, 郭卉欣, 黄新春, 陈燕梅, 魏文静()   

  1. 510630 广州,广东省结核病控制中心结核病研究所(张晨晨、郭卉欣、黄新春、陈燕梅、魏文静);深圳市慢性病防治中心肺部疾病防治研究所(谭卫国)
  • 收稿日期:2020-10-23 出版日期:2021-04-10 发布日期:2021-04-09
  • 通信作者: 张晨晨,魏文静 E-mail:wenjingwei2014@163.com
  • 基金资助:
    “十三五”国家科技重大专项(2018ZX10715004-002-021);“十三五”国家科技重大专项(2017ZX10201302-005);广东省医学科学技术研究基金(A2020338)

The effect of Mycobacterium tuberculosis infection on sputum flora structure

ZHANG Chen-chen*, TAN Wei-guo, GUO Hui-xin, HUANG Xin-chun, CHEN Yan-mei, WEI Wen-jing()   

  1. *Research Institute of Tuberculosis, Centre for Tuberculosis Control of Guangdong Province, Guangzhou 510630, China
  • Received:2020-10-23 Online:2021-04-10 Published:2021-04-09
  • Contact: ZHANG Chen-chen*,WEI Wen-jing E-mail:wenjingwei2014@163.com

摘要:

目的 探索健康人、结核分枝杆菌潜伏感染者和活动性肺结核患者痰液菌群的差异。方法 搜集2016年11月至2017年12月深圳市慢性病防治中心健康体检者53名作为A组,登记确诊的结核分枝杆菌潜伏感染(LTBI)者41例作为B组,初治活动性肺结核患者54例作为C组。研究对象均于清晨空腹状态下收集下呼吸道痰液样本(以干酪痰和黏液痰为佳)3~5ml,共148份。痰液样本提取总DNA,对16S rRNA V4~V5区基因进行高通量测序,并对测序结果进行生物信息学分析。结果 通过对148份痰液样本进行高通量测序分析,获得有效序列14136502.0条,获得操作分类单元(operational taxonomic units,OTU)21712.00个。Alpha多样性分析显示,A组Pielou_e指数为0.85(0.82,0.88),明显高于C组[0.83(0.78,0.86)],差异有统计学意义(H=4.462,P=0.035)。Beta多样性分析发现,三组人群痰液菌群组成存在差异(H=2.027,P=0.002)。菌群组间差异物种分析发现,三组人群在3个门(厚壁菌门、梭杆菌门、螺旋体门)、3个纲(黄杆菌纲、梭杆菌纲、螺旋体纲)、5个目(黄杆菌目、乳杆菌目、梭杆菌目、伯克霍尔德菌目、螺旋体目)、6个科(紫单胞菌科、黄杆菌科、梭杆菌科、纤毛菌科、伯克菌科、螺旋体科)、5个属(二氧化碳噬纤维菌属、卟啉单胞菌属、梭杆菌属、纤毛菌属、密螺旋体属)和2个种(NanceiensisParvula)水平上均存在差异;其中,C组中伯克霍尔德菌目(Burkholderiales)[0.01(0.00,0.02)]和伯克菌科(Burkholderiaceae)[0.01(0.00,0.01)]的丰富度较A组[丰富度分别为0.00(0.00,0.01)和0.00(0.00,0.00)]和B组[丰富度分别为0.00(0.00,0.01)和0.00(0.00,0.00)]明显增加,差异均有统计学意义(C组与A组比较:H值分别为9.733和4.799,P值分别为0.002和0.028;C组与B组比较:H值分别为12.134和8.152,P值均<0.01)。结论 结核分枝杆菌感染未改变人痰液菌群的丰富度,但造成菌群结构组成上产生差异。

关键词: 分枝杆菌,结核, 痰, 细菌结构, 生物多样性, 对比研究

Abstract:

Objective To explore the difference of sputum flora in healthy population, latent tuberculosis infection (LTBI) and active pulmonary tuberculosis patients. Methods From November 2016 to December 2017, 53 healthy subjects from Shenzhen Center of Chronic Diseases Prevention and Control were enrolled as group A, 41 registered patients with confirmed LTBI were enrolled as group B, and 54 patients with newly treated active tuberculosis were enrolled as group C. The 148 sputum samples of the lower respiratory tract (cheese sputum and mucus sputum were preferred) were collected from all the subjects in the morning when limosis. Total DNA was extracted from sputum samples, and 16S rRNA V4-V5 region genes were sequenced by high-throughput sequencing, the sequencing results were analyzed by bioinformatics. Results Through high-throughput sequencing analysis of the 148 sputum samples, 14136502.0 effective sequences and 21712.00 operational taxonomic units (OTU) were obtained. Alpha diversity analysis showed that the Pielou_e index of group A was 0.85 (0.82, 0.88), which was significantly higher than that of group C (0.83 (0.78, 0.86), H=4.462, P=0.035). Beta diversity analysis showed that there were differences in sputum flora composition among the three groups (H=2.027, P=0.002). Species analysis of different bacterial groups showed significant differences in levels of different Phylum (Firmicutes, Fusobacteria, Spirochaetes), Class (Flavobacteriia, Fusobacteriia, Spirochaetes), Order (Flavobacteriales, Lactobacillales, Fusobacteriales, Burkholderiales, Spirochaetales), Family (Porphyromonadaceae, Flavobacteriaceae, Fusobacteriaceae, Leptotrichiaceae, Burkholderiaceae, Spirochaetaceae), Genus (Capnocytophaga, Porphyromonas, Fusobacterium, Leptotrichia, Treponema) and Species (Nanceiensis,Parvula) among the three groups. Of them, the richness of Burkholderiales (0.01 (0.00, 0.02)) and Burkholderiaceae (0.01 (0.00, 0.01)) in group C was significantly higher than those in group A (0.00 (0.00, 0.01) and 0.00 (0.00, 0.00), respectively) and group B (0.00 (0.00, 0.01) and 0.00 (0.00, 0.00), respectively) (group C vs. group A: H values were 9.733 and 4.799, P values were 0.002 and 0.028, respectively; group C vs. group B: H values were 12.134 and 8.152, respectively, P values were both <0.01). Conclusion Mycobacterium tuberculosis infection did not change the richness of human sputum bacteria, however, it caused differences in the composition of the bacterial community.

Key words: Mycobacterium tuberculosis, Sputum, Bacterial structures, Biodiversity, Comparative study