Department of Thoracic Medicine, Haukeland University Hospital, Bergen, Norway.
Department of Clinical Science, Faculty of Medicine and Dentistry, University of Bergen, Bergen, Norway.
Respir Res. 2017 Aug 29;18(1):164. doi: 10.1186/s12931-017-0645-3.
Induced and spontaneous sputum are used to evaluate the airways microbiota. Whether the sputum types can be used interchangeably in microbiota research is unknown. Our aim was to compare microbiota in induced and spontaneous sputum from COPD patients sampled during the same consultation.
COPD patients from Bergen, Norway, were followed between 2006/2010, examined during the stable state and exacerbations. 30 patients delivered 36 sample pairs. DNA was extracted by enzymatic and mechanical lysis methods. The V3-V4 region of the 16S rRNA gene was PCR-amplified and prepared for paired-end sequencing. Illumina Miseq System was used for sequencing, and Quantitative Insights Into Microbial Ecology (QIIME) and Stata were used for bioinformatics and statistical analyses.
Approximately 4 million sequences were sorted into 1004 different OTUs and further assigned to 106 different taxa. Pair-wise comparison of both taxonomic composition and beta-diversity revealed significant differences in one or both parameters in 1/3 of sample pairs. Alpha-diversity did not differ. Comparing abundances for each taxa identified, showed statistically significant differences between the mean abundances in induced versus spontaneous samples for 15 taxa when disease state was considered. This included potential pathogens like Haemophilus and Moraxella.
When studying microbiota in sputum samples one should take into consideration how samples are collected and avoid the usage of both induced and spontaneous sputum in the same study.
诱导痰和自发性痰常用于评估气道微生物群。在微生物群研究中，这两种痰标本是否可以互换使用尚不清楚。我们的目的是比较在同一次就诊期间采集的 COPD 患者的诱导痰和自发性痰中的微生物群。
挪威卑尔根的 COPD 患者在 2006/2010 年期间接受随访，在稳定期和加重期进行检查。30 名患者提供了 36 对样本。通过酶和机械裂解方法提取 DNA。PCR 扩增 16S rRNA 基因的 V3-V4 区，并准备进行双端测序。使用 Illumina Miseq 系统进行测序，使用 Quantitative Insights Into Microbial Ecology (QIIME) 和 Stata 进行生物信息学和统计分析。
大约 400 万个序列被分为 1004 个不同的 OTUs，并进一步分配到 106 个不同的分类群。通过对分类组成和β多样性进行两两比较，在 1/3 的样本对中发现了一个或两个参数存在显著差异。α多样性没有差异。当考虑疾病状态时，比较每个分类群的丰度，发现 15 个分类群的诱导样本与自发性样本的平均丰度存在统计学显著差异，其中包括潜在的病原体如嗜血杆菌和莫拉菌。
在研究痰样本中的微生物群时，应考虑到样本的采集方式，并避免在同一研究中同时使用诱导痰和自发性痰。
