Department of Respiratory Medicine, University of Occupational and Environmental Health, Japan, Kitakyushu, Fukuoka, Japan.
Department of Microbiology, University of Occupational and Environmental Health, Japan, Kitakyushu, Fukuoka, Japan.
Respirology. 2015 Jul;20(5):758-65. doi: 10.1111/resp.12536. Epub 2015 Mar 30.
Recent advances in cultivation-independent molecular biological modalities for detecting bacterial species have indicated that several bacterial species may play a role in the pathogenesis of certain infectious diseases. The aim of this study was to evaluate the role of bacterial flora in the pathogenesis of nontuberculous mycobacteriosis (NTM) using a bacterial floral analysis of bronchoalveolar lavage fluid (BALF) with 16S rRNA gene sequencing in patients with bronchiectasis.
Fifty-eight patients with bronchiectasis evaluated using chest computed tomography were enrolled. BALF obtained from the most affected lung lesions was evaluated using culture and culture-independent methodologies. Approximately 600 bp of the bacterial 16S rRNA gene (E341F-E907R) was amplified via polymerase chain reaction using universal primers, and clone libraries were constructed. Nucleotide sequences of 96 randomly chosen clones for each specimen were determined, and the homology was searched using a basic local alignment search tool to determine the bacterial phylotypes and their proportions (bacterial floral analysis) in each specimen.
Twenty-nine patients with bronchiectasis were diagnosed with NTM based on culture-based methods using Ogawa medium. The molecular method showed a significantly high rate of anaerobes among the patients with NTM compared with that observed in the bronchiectasis patients without NTM. In addition, findings of collapse/consolidation were significantly related to the proportion of Prevotella species in the BALF samples determined using the molecular method (P < 0.001).
Given the results of the present study, anaerobes may play an important role in the pathogenesis of bronchiectasis in patients with NTM.
最近在检测细菌物种的非培养依赖性分子生物学方法方面取得了进展，这表明几种细菌可能在某些传染病的发病机制中发挥作用。本研究旨在使用支气管肺泡灌洗液（BALF）中的细菌菌群分析，通过 16S rRNA 基因测序来评估细菌菌群在支气管扩张症患者中非结核分枝杆菌病（NTM）发病机制中的作用。
对 58 例经胸部计算机断层扫描评估的支气管扩张症患者进行了研究。使用培养和非培养方法评估从最受影响的肺病变中获得的 BALF。通过聚合酶链反应使用通用引物扩增细菌 16S rRNA 基因（E341F-E907R）的约 600 bp，并构建克隆文库。对每个标本的 96 个随机选择的克隆的核苷酸序列进行测定，并使用基本局部比对搜索工具搜索同源性，以确定每个标本中的细菌型和其比例（细菌菌群分析）。
根据使用 Ogawa 培养基的基于培养的方法，29 例支气管扩张症患者被诊断为 NTM。与支气管扩张症无 NTM 的患者相比，分子方法显示 NTM 患者的厌氧菌比例明显较高。此外，使用分子方法确定的 BALF 样本中普雷沃氏菌属比例与塌陷/实变的发现显著相关（P<0.001）。
鉴于本研究的结果，厌氧菌可能在 NTM 患者支气管扩张症的发病机制中发挥重要作用。
