声带良性增生性疾病患者声带表面分泌物菌群研究
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摘要
目的比较声带良性增生性疾病患者与正常人声带表面分泌物中菌群组成的异同。方法以声带良性增生性疾病患者32例为实验组,33例声带正常的甲状腺肿物患者作为正常对照组,分别采集两组对象声带表面分泌物并提取细菌基因组DNA,经过对16S rRNA V3-V4区的PCR扩增后,用Illumina平台进行二代DNA测序,通过与Greengenes数据库进行物种比对及分类,在菌门水平比较实验组和对照组之间菌落构成的异同。结果两组检测到的微生物丰度差异无统计学意义(P=0.531);两组中占前7位的菌门都是变形菌门(proteobacteria)、厚壁菌门(firmicutes)、拟杆菌门(bacteroidetes)、梭杆菌门(fusobacteria)、放线菌门(actinobacteria)、SR1和TM7,实验组中变形菌门、厚壁菌门所占比例明显高于对照组(分别为P<0.001、P<0.01),而拟杆菌门、梭杆菌门、放线菌门及SR1所占比例较对照组明显减少(分别为P<0.001、P<0.01、P<0.05及P<0.01),差异均有统计学意义。TM7在实验组和对照组中分别占1.08%±0.20%和1.61%±0.21%,差异无统计学意义(P>0.05)。结论在菌门水平,声带良性增生性疾病患者声带表面细菌数量和优势菌群与正常人相似,但各菌门在两组中所占比例存在显著差异,这些菌门构成上的差异可能与声带良性增生性疾病的发生、发展有关。
Objective To compare the microbial communities of vocal-folds between patients with benign vocal-fold lesions and normal people. Methods From Jun 2015 to Dec 2015, 32 patients with benign vocal fold lesions in the ENT clinic of Renmin Hospital of Wuhan University were enrolled into the experimental group, with another 33 people diagnosed with thyroid neoplasms as control. Secreta samples from vocal-folds were collected for bacterial genomic DNA extraction. After PCR amplification targeting on the V3-V4 divisions of 16 s ribosomal RNA and second generation DNA sequencing performed on illumina platform, the operational taxnomic units(OTUs) were compared to those in Greengenes database. The microbial communities were analysed at phylum level.Results There was no significant difference of average bacterial amount between these two groups(P=0.531). The top 7 phyla were proteobacteria, firmicutes, bacteroidetes, fusobacteria, actinobacteria, SR1 and TM7. The proportions of proteobacteria and firmicutes were significantly higher in benign vocal-fold lesion group than those in the control group, while the proportions of bacteroidetes, fusobacteria, actinobacteria and SR1 were significantly reduced in the benign vocal-fold lesion group. The proportion of TM7 was 1.08%±0.20% in benign vocal-fold lesion group, and 1.61%±0.21% in control group. However, there was no significant difference between two groups.Conclusion The bacterial amount and dominant flora on the vocal fold surface from benign vocal fold lesion patients are similar to those from normal people at the phylum level. However, the percentages of each phylum are significantly different in these two groups, which may be associated with the underlying physiopathology of the benign vocal fold lesions.
Objective To compare the microbial communities of vocal-folds between patients with benign vocal-fold lesions and normal people. Methods From Jun 2015 to Dec 2015, 32 patients with benign vocal fold lesions in the ENT clinic of Renmin Hospital of Wuhan University were enrolled into the experimental group, with another 33 people diagnosed with thyroid neoplasms as control. Secreta samples from vocal-folds were collected for bacterial genomic DNA extraction. After PCR amplification targeting on the V3-V4 divisions of 16 s ribosomal RNA and second generation DNA sequencing performed on illumina platform, the operational taxnomic units(OTUs) were compared to those in Greengenes database. The microbial communities were analysed at phylum level.Results There was no significant difference of average bacterial amount between these two groups(P=0.531). The top 7 phyla were proteobacteria, firmicutes, bacteroidetes, fusobacteria, actinobacteria, SR1 and TM7. The proportions of proteobacteria and firmicutes were significantly higher in benign vocal-fold lesion group than those in the control group, while the proportions of bacteroidetes, fusobacteria, actinobacteria and SR1 were significantly reduced in the benign vocal-fold lesion group. The proportion of TM7 was 1.08%±0.20% in benign vocal-fold lesion group, and 1.61%±0.21% in control group. However, there was no significant difference between two groups.Conclusion The bacterial amount and dominant flora on the vocal fold surface from benign vocal fold lesion patients are similar to those from normal people at the phylum level. However, the percentages of each phylum are significantly different in these two groups, which may be associated with the underlying physiopathology of the benign vocal fold lesions.
引文
1 Cohen SM,Dupont WD,Courey MS.Quality-of-life impact of nonneoplastic voice disorders:a meta-analysis[J].Ann Otol Rhinol Laryngol,2006,115:128.
2 The human microbiome project consortium:a framework for human microbiome research[J].Nature,2012,doi:10.1038/ nature11209.
3 Klindworth A,Pruesse E,Schweer T.Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies[J].Nucleic Acids Res,2013,41:e1.
4 Mao JP,Wang QH,Zhou Y.Bridge PCR,an easy way for concatemerizing DNA tags[J].China Biotechnology,2009,29:66.
5 Kuczynski J,Stombaugh J,Walters WA.Using QIIME to analyze 16S rRNA gene sequences from microbial communities[J].Curr Protoc Bioinformatics,2011,10:1.
6 Savage DC.Microbial ecology of the gastrointestinal tract[J].Annu Rev Microbiol,1977,31:107.
7 Kleinberg I.A mixed-bacteria ecological approach to understanding the role of the oral bacteria in dental caries causation:an alternative to streptococcus mutans and the specific-plaque hypothesis[J].Crit Rev Oral Biol Med,2002,13:108.
8 Wade WG.The oral microbiome in health and disease[J].Pharmacological Research,2013,69:137.
9 Gong HL,Shi Y,Zhou X,et al.Microbiota in the throat and risk factors for laryngeal carcinoma[J].AEM,2014,80:7356.
10 Hanshew AS.Characterization and comparison of bacterial communities in benign vocal fold lesions[J].Microbiome,2014,2:43.
11 Gursoy UK,Kononen E,Uitto VJ.Prevotella intermedia ATCC 25611 targets host cell lamellipodia in epithelial cell adhesion and invasion[J].Oral Microbiol Immunol,2009,24:304.
12 Jacobsen UP,Nielsen HB,Hildebrand F.The chemical interactome space between the human host and the genetically defined gut metabotypes[J].ISME J,2013,7:730.
13 Dueholm MS,Albertsen M,Stokholm-Bjerregaard M.Complete genome sequence of the bacterium Aalborg_AAW-1,representing a novel family within the candidate phylum SR1[J].Genome Announc,2015,3:e00624.
14 Marcy Y,Ouverney C,Bik EM.Dissecting biological “dark matter” with single-cell genetic analysis of rare and uncultivated TM7 microbes from the human mouth[J].Proc Natl Acad Sci USA,2007,104:11889.
2 The human microbiome project consortium:a framework for human microbiome research[J].Nature,2012,doi:10.1038/ nature11209.
3 Klindworth A,Pruesse E,Schweer T.Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies[J].Nucleic Acids Res,2013,41:e1.
4 Mao JP,Wang QH,Zhou Y.Bridge PCR,an easy way for concatemerizing DNA tags[J].China Biotechnology,2009,29:66.
5 Kuczynski J,Stombaugh J,Walters WA.Using QIIME to analyze 16S rRNA gene sequences from microbial communities[J].Curr Protoc Bioinformatics,2011,10:1.
6 Savage DC.Microbial ecology of the gastrointestinal tract[J].Annu Rev Microbiol,1977,31:107.
7 Kleinberg I.A mixed-bacteria ecological approach to understanding the role of the oral bacteria in dental caries causation:an alternative to streptococcus mutans and the specific-plaque hypothesis[J].Crit Rev Oral Biol Med,2002,13:108.
8 Wade WG.The oral microbiome in health and disease[J].Pharmacological Research,2013,69:137.
9 Gong HL,Shi Y,Zhou X,et al.Microbiota in the throat and risk factors for laryngeal carcinoma[J].AEM,2014,80:7356.
10 Hanshew AS.Characterization and comparison of bacterial communities in benign vocal fold lesions[J].Microbiome,2014,2:43.
11 Gursoy UK,Kononen E,Uitto VJ.Prevotella intermedia ATCC 25611 targets host cell lamellipodia in epithelial cell adhesion and invasion[J].Oral Microbiol Immunol,2009,24:304.
12 Jacobsen UP,Nielsen HB,Hildebrand F.The chemical interactome space between the human host and the genetically defined gut metabotypes[J].ISME J,2013,7:730.
13 Dueholm MS,Albertsen M,Stokholm-Bjerregaard M.Complete genome sequence of the bacterium Aalborg_AAW-1,representing a novel family within the candidate phylum SR1[J].Genome Announc,2015,3:e00624.
14 Marcy Y,Ouverney C,Bik EM.Dissecting biological “dark matter” with single-cell genetic analysis of rare and uncultivated TM7 microbes from the human mouth[J].Proc Natl Acad Sci USA,2007,104:11889.