基于高通量测序的分析：幽门螺杆菌感染与唾液微生物组。
Analysis of by high-throughput sequencing: Helicobacter pylori infection and salivary microbiome.
机构信息
Division of Gastroenterology and Hepatology, Key Laboratory of Gastroenterology and Hepatology, Ministry of Health, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
出版信息
BMC Oral Health. 2020 Mar 20;20(1):84. doi: 10.1186/s12903-020-01070-1.
BACKGROUND
There have been reports of Helicobacter pylori (H. pylori) in the oral cavity and it has been suggested that the oral cavity may be a reservoir for H. pylori reflux from the stomach. High-throughput sequencing was used to assess the structure and composition of oral microbiota communities in individuals with or without confirmed H. pylori infection.
METHODS
Saliva samples were obtained from 34 H. pylori infected and 24 H. pylori uninfected subjects. Bacterial genomic DNA was extracted and examined by sequencing by amplification of the 16S rDNA V3-V4 hypervariable regions followed by bioinformatics analysis. Saliva sampling was repeated from 22 of the 34 H. pylori infected subjects 2 months after H. pylori eradication.
RESULTS
High-quality sequences (2,812,659) clustered into 95,812 operational taxonomic units (OTUs; 97% identity). H. pylori was detected in the oral cavity in infected (12/34), uninfected (11/24) and eradicated (15/22) subjects by technique of high-throughput sequencing, occupying 0.0139% of the total sequences. Alpha diversity of H. pylori infected subjects was similar to that of uninfected subjects (Shannon: 1417.58 vs. 1393.60, p > 0.05, ACE: 1491.22 vs. 1465.97, p > 0.05, Chao 1: 1417.58 vs. 1393.60, p > 0.05, t-test). Eradication treatment decreased salivary bacterial diversity (Shannon, p = 0.015, ACE, p = 0.003, Chao 1, p = 0.002, t-test). Beta diversity analysis based on unweighted UniFrac distances showed that the salivary microbial community structure differed between H. pylori infected and uninfected subjects (PERMANOVAR, pseudo-F: 1.49, p = 0.033), as well as before and after H. pylori eradication (PERMANOVAR, pseudo-F: 3.34, p = 0.001). Using LEfSe analysis, 16 differentially abundant genera were defined between infected and uninfected subjects, 12 of which had a further alteration after successful eradication.
CONCLUSIONS
Our study using high-throughput sequencing showed that H. pylori was present commonly in the oral cavity with no clear relation to H. pylori infection of the stomach. Both H. pylori infection and eradication therapy caused alterations in community and structure of the oral microbiota.
TRIAL REGISTRATION
clinicaltrials.gov, NCT03730766. Registered 2 Nov 2018 - Retrospectively registered, https://clinicaltrials.gov/ct2/show/ NCT03730766.
背景
已有报道称幽门螺杆菌（H. pylori）存在于口腔中，有人认为口腔可能是 H. pylori 从胃反流的储库。本研究采用高通量测序技术评估了经证实感染 H. pylori 和未感染 H. pylori 的个体的口腔微生物群落结构和组成。
方法
从 34 例 H. pylori 感染和 24 例 H. pylori 未感染的受试者中采集唾液样本。提取细菌基因组 DNA，通过扩增 16S rDNA V3-V4 高变区进行测序，然后进行生物信息学分析。在 H. pylori 根除后 2 个月，对 34 例 H. pylori 感染患者中的 22 例重复进行唾液采样。
结果
高质量序列（2812659 个）聚类为 95812 个操作分类单元（OTUs；97%的同一性）。通过高通量测序技术，在感染（12/34）、未感染（11/24）和根除（15/22）的受试者口腔中均检测到 H. pylori，占总序列的 0.0139%。感染 H. pylori 的受试者的 alpha 多样性与未感染的受试者相似（香农：1417.58 与 1393.60，p>0.05；ACE：1491.22 与 1465.97，p>0.05；Chao 1：1417.58 与 1393.60，p>0.05，t 检验）。根除治疗降低了唾液细菌多样性（香农，p=0.015；ACE，p=0.003；Chao 1，p=0.002，t 检验）。基于非加权 UniFrac 距离的 beta 多样性分析表明，H. pylori 感染和未感染受试者的唾液微生物群落结构不同（PERMANOVAR，伪 F：1.49，p=0.033），以及 H. pylori 根除前后（PERMANOVAR，伪 F：3.34，p=0.001）。使用 LEfSe 分析，确定了感染和未感染受试者之间 16 个差异丰度的属，其中 12 个在成功根除后有进一步的变化。
结论
本研究采用高通量测序技术表明，H. pylori 通常存在于口腔中，与 H. pylori 胃部感染无明显关系。H. pylori 感染和根除治疗均导致口腔微生物群落结构发生改变。
试验注册
clinicaltrials.gov，NCT03730766。注册日期：2018 年 11 月 2 日-回顾性注册，https://clinicaltrials.gov/ct2/show/NCT03730766。
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