Detection of bacterial DNA in lymph nodes of Crohn's disease patients using high throughput sequencing
Detection of bacterial DNA in lymph nodes of Crohn's disease patients using high throughput sequencing
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DOI：
10.1136/gutjnl-2013-305320
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发表时间：
2014-10-01
期刊：
影响因子：
24.5
通讯作者：
Allison, Gwen E.
中科院分区：
文献类型：
--
作者：
O'Brien, Claire L.;Pavli, Paul;Allison, Gwen E.
目的通过比较克罗恩病（Crohn 'sdisease，CD）患者和对照组淋巴结和粘膜的微生物群落，探讨CD患者淋巴结中是否存在特异性的微生物转运。设计淋巴结，并参与和未参与的粘膜样品，从58例（29 CD，8个“其他炎症性肠病”（IBD）和21个非IBD）的切除获得。使用靶向细菌16 S rRNA基因的V1-V3区域的通用引物扩增细菌DNA，并使用高通量测序对扩增子进行测序。20例患者（CD 8例（28%），其他IBD 2例（25%），非IBD 10例（48%））有PCR阳性淋巴结。所有CD样本的淋巴结中均不存在特异性微生物。大肠埃希菌/志贺氏菌在所有患者组中均很常见，但回肠CD患者的淋巴结中大肠埃希菌读数比例高于其他CD患者（p=0.0475）。弯曲杆菌，螺杆菌和耶尔森菌是罕见的;分枝杆菌和李斯特菌未检测到。菌群失调是存在于所有组，但变化是具体的，没有共同的模式emerging.Conclusions它是不可能的，一个单一的细菌在晚期CD炎症永存;菌群失调是常见的，我们没有发现增加细菌易位的证据。我们认为，未来的研究应该集中在早期疾病和活菌在节点，阿弗他溃疡和肉芽肿，因为它们可能是更相关的炎症CD的启动。
Objective Our aim was to determine whether or not specific microorganisms were transported selectively to lymph nodes in Crohn's disease (CD) by comparing node and mucosal microbial communities in patients and controls. We also sought evidence of dysbiosis and bacterial translocation.Design Lymph nodes, and involved and uninvolved mucosal samples were obtained from resections of 58 patients (29 CD, eight 'other inflammatory bowel disease' (IBD) and 21 non-IBD). Universal primers targeting V1-V3 regions of bacterial 16S rRNA genes were used to amplify bacterial DNA and amplicons sequenced using high throughput sequencing. 20 patients (eight CD (28%), two other IBD (25%) and 10 non-IBD (48%)) had PCR positive nodes.Results All samples from an individual were similar: there was no evidence of selective concentration of any microorganism in nodes. No specific microorganism was present in the nodes of all CD samples. Escherichia/Shigella were common in all patient groups but patients with ileal CD had a greater proportion of Escherichia coli reads in their nodes than other CD patients (p=0.0475). Campylobacter, Helicobacter and Yersinia were uncommon; Mycobacterium and Listeria were not detected. Dysbiosis was present in all groups but shifts were specific and no common pattern emerged.Conclusions It is unlikely that a single bacterium perpetuates inflammation in late stage CD; dysbiosis was common and we found no evidence of increased bacterial translocation. We believe that future studies should focus on early disease and viable bacteria in nodes, aphthous ulcers and granulomas, as they may be more relevant in the initiation of inflammation in CD.