Diarrhoea-predominant irritable bowel syndrome distinguishable by 16S rRNA gene phylotype quantification
Diarrhoea-predominant irritable bowel syndrome distinguishable by 16S rRNA gene phylotype quantification
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DOI：
10.3748/wjg.15.5936
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发表时间：
2009-12-21
影响因子：
4.3
通讯作者：
Palva, Airi
中科院分区：
文献类型：
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作者：
Lyra, Anna;Rinttila, Teemu;Palva, Airi
目的：研究选定的细菌16S核糖体RNA（rRNA）基因系统型是否能够区分肠易激综合征（IBS）。
方法：将20名IBS志愿者（分为8名腹泻型（IBS - D）、8名便秘型（IBS - C）和4名混合症状亚型（IBS - M）IBS患者）以及15名对照受试者的粪便微生物群在三个时间点用一组14种定量实时聚合酶链反应测定法进行分析。所有测定均基于16S rRNA基因文库序列分析，针对假定与IBS相关的16S rRNA基因系统型。目标系统型隶属于放线菌门、拟杆菌门和厚壁菌门。根据其16S rRNA基因序列，8种目标系统型与已培养的细菌物种相似度低于95%。数据分析采用重复测量方差分析类型的数据建模以及主成分分析（PCA），并将线性混合效应模型应用于主成分得分。
结果：从所有分析的样本中检测到细菌系统型球形梭菌88%、热琥珀酸梭菌85%、链状粪杆菌91%、类瘤胃球菌、扭链瘤胃球菌91%以及扭链瘤胃球菌93%。对所有14种细菌16S rRNA基因系统型相对数量的多变量分析表明，IBS - D患者的肠道微生物群与其他样本组不同。对第一个主成分（PC1）的PCA分析（解释了IBS - D患者组中观察到的30.36%的变异）与所有其他样本组有显著差异（IBS - D与对照组，P = 0.01；IBS - D与IBS - M组，P = 0.00；IBS - D与IBS - C组，P = 0.05）。使用与细菌总量成比例的相对值时，在不同系统型的水平上也观察到显著差异。一种与热琥珀酸梭菌相似度为85%的系统型在IBS - D患者和对照组之间的数量有显著差异（-4.08 ± 0.90与 -3.33 ± 1.16，P = 0.04），在IBS - D患者和IBS - M患者之间也有显著差异（-4.08 ± 0.90与 -3.08 ± 1.38，P = 0.05）。此外，一种与扭链瘤胃球菌相似度为94%的系统型在IBS - D患者的肠道微生物群中比在对照组中更普遍（-2.43 ± 1.49与 -4.02 ± 1.63，P = 0.01）。一种与扭链瘤胃球菌相似度为93%的系统型与对照组样本相关，而与IBS - M组相比（-2.41 ± 0.53与 -2.92 ± 0.56，P = 0.00）。另外，一种类瘤胃球菌系统型与IBS - C患者相关，而与对照组相比（-1.61 ± 1.83与 -3.69 ± 2.42，P = 0.01）。上述所有系统型特异性的改变均与时间的影响无关。
结论：观察到IBS患者肠道微生物群在系统型水平上有显著改变，进一步强调了胃肠道微生物群在IBS中的可能作用。（C）2009世界胃肠病学杂志出版社和百世登。保留所有权利。
AIM: To study whether selected bacterial 16S ribosomal RNA (rRNA) gene phylotypes are capable of distinguishing irritable bowel syndrome (IBS).METHODS: The faecal microbiota of twenty volunteers with IBS, subdivided into eight diarrhoea-predominant (IBS-D), eight constipation-predominant (IBS-C) and four mixed symptom-subtype (IBS-M) IBS patients, and fifteen control subjects, were analysed at three time-points with a set of fourteen quantitative real-time polymerase chain reaction assays. All assays targeted 16S rRNA gene phylotypes putatively associated with IBS, based on 16S rRNA gene library sequence analysis. The target phylotypes were affiliated with Actinobacteria, Bacteroidetes and Firmicutes. Eight of the target phylotypes had less than 95% similarity to cultured bacterial species according to their 16S rRNA gene sequence. The data analyses were made with repeated-measures ANCOVA-type modelling of the data and principle component analysis (PCA) with linear mixed-effects models applied to the principal component scores.RESULTS: Bacterial phylotypes Clostridium cocleatum 88%, Clostridium thermosuccinogenes 85%, Coprobacillus catenaformis 91%, Ruminococcus bromii-like, Ruminococcus torques 91%, and R. torques 93% were detected from all samples analysed. A multivariate analysis of the relative quantities of all 14 bacterial 16S rRNA gene phylotypes suggested that the intestinal microbiota of the IBS-D patients differed from other sample groups. The PCA on the first principal component (PC1), explaining 30.36% of the observed variation in the IBS-D patient group, was significantly altered from all other sample groups (IBS-D vs control, P = 0.01; IBS-D vs IBS-M, P = 0.00; IBS-D vs IBS-C, P = 0.05). Significant differences were also observed in the levels of distinct phylotypes using relative values in proportion to the total amount of bacteria. A phylotype with 85% similarity to C thermosuccinogenes was quantified in significantly different quantities among the IBS-D and control subjects (-4.08 +/- 0.90 vs -3.33 +/- 1.16, P = 0.04) and IBS-D and IBS-M subjects (-4.08 +/- 0.90 vs -3.08 +/- 1.38, P = 0.05). Furthermore, a phylotype with 94% similarity to R. torques was more prevalent in IBS-D patients' intestinal microbiota than in that of control subjects (-2.43 +/- 1.49 vs -4.02 +/- 1.63, P = 0.01). A phylotype with 93% similarity to R. torques was associated with control samples when compared with IBS-M (-2.41 +/- 0.53 vs -2.92 +/- 0.56, P = 0.00). Additionally, a R. bromii-like phylotype was associated with IBS-C patients in comparison to control subjects (-1.61 +/- 1.83 vs -3.69 +/- 2.42, P = 0.01). All of the above mentioned phylotype specific alterations were independent of the effect of time.CONCLUSION: Significant phylotype level alterations in the intestinal microbiotas of IBS patients were observed, further emphasizing the possible contribution of the gastrointestinal microbiota in IBS. (C) 2009 The WJG Press and Baishideng. All rights reserved.