引用本文:施春梅,周 嫣,方邦骞,欧晓丽,谢庆玲,钟秀芬.PCR-DGGE技术应用于手足口病儿童口腔微生物菌群变化的初步研究[J].中国临床新医学,2015,8(2):97-101.
【打印本页】   【下载PDF全文】   查看/发表评论  【EndNote】   【RefMan】   【BibTex】
←前一篇|后一篇→ 过刊浏览    高级检索
本文已被:浏览 1353次   下载 1071 本文二维码信息
码上扫一扫!
分享到: 微信 更多
PCR-DGGE技术应用于手足口病儿童口腔微生物菌群变化的初步研究
施春梅,周 嫣,方邦骞,欧晓丽,谢庆玲,钟秀芬
530021 南宁,广西壮族自治区人民医院口腔科
摘要:
[摘要] 目的 应用PCR-DGGE技术对手足口病(HFMD)、疱疹性口腔炎(HGS)及正常儿童口腔微生物菌群变化进行初步分析,以期了解三者是否存在差异。方法 按随机数字表法选择3例手足口病唾液样本,1例幼儿园正常儿童唾液以及1例疱疹性口腔炎患儿唾液。样本进行微生物群落总DNA的抽提;以此为模板扩增16S rRNA V3可变区,产物经DGGE指纹图谱分析其组成结构,并运用UVIBAND/MAP等软件比较所得群落指纹图谱的相似性指数;对DGGE指纹图谱上的泳带中优势条带切胶测序,进行基因序列分析,得出该样本的优势菌群。结果 手足口病组优势菌群:Capnocytophaga嗜二氧化碳噬细胞菌、Selenomonas月形单胞菌属、Prevotella普氏菌属、TM7_genera_incertae_sedis菌属 、Streptococcus链球菌属、Neisseria奈瑟菌属、Porphyromonas卟啉单胞菌属、Campylobacter弯曲菌属、Campylobacter弯曲菌属、Granulicatella 菌属、Leptotrichia 纤毛菌属、Veillonella韦永氏球菌属、Bacteroides-like sp菌属、Actinomyces放线菌、Fusobacterium梭菌属。正常对照组优势菌群:Veillonella韦永氏球菌属、Capnocytophaga嗜二氧化碳噬细胞菌、Prevotella普氏菌、Gemella双胎溶血菌属、Porphyromonas卟啉单胞菌属、Ruminococcaceae瘤胃球菌属、Lachnospiraceae毛螺菌属、Selenomonas月形单胞菌属、Clostridium梭状芽孢杆菌、Haemophilus嗜血杆菌。疱疹性口腔炎组优势菌群:Lactococcus乳球菌属、Prevotella普氏菌属。结论 手足口病组菌群结构更为复杂,与疱疹性口腔炎组相比菌群结构差异明显。
关键词:  PCR-DGGE技术  手足口病  疱疹性口腔炎
DOI:10.3969/j.issn.1674-3806.2015.02.01
分类号:R 788
基金项目:广西自然科学基金资助项目(编号:2011GXNSFA018252)
A preliminary application of PCR-DGGE technology to research the changes of oral microbial floras in children with hand foot and mouth disease
SHI Chun-mei, ZHOU Yan, FANG Bang-qian, et al.
Department of Dentistry, the People′s Hospital of Guangxi Zhuang Autonomous Region, Nanning 530021, China
Abstract:
[Abstract] Objective To apply PCR-DGGE technology analysing changes of oral microbial floras in children with hand foot and mouth disease(HFMD).Methods According to the random number table method,3 children with HFMD were randomly chosen and their saliva samples were collected;at the same time, saliva samples were collected from one normal child,and from one child with herpetic ginggivostomatitis(HGS). Samples′ total DNA extraction of the microbial community were performed; taking it as template for amplification of 16 s rRNA V3 variable area, the structure of product by the DGGE fingerprint, and using UVIBAND/MAP software for comparing similarity index of the community fingerprint; the edge banding on swimming belt of DGGE fingerprint were excised from the gel for sequencing, gene sequence analysis was performed, then the sample advantage bacterium group was obtained.Results The advantage bacterial colonies of HFMD group: Capnocytophaga, Selenomonas, Prevotella, TM7_genera_incertae_sedis, Streptococcus,Neisseria,Porphyromonas,Campylobacter,Campylobacter,Granulicatell-a, Leptotrichia, Veillonella, Bacteroides-like sp, Actinomyces, Fusobacterium. The advantage bacterial colonies of normal group: Veillonella, Capnocytophaga, Prevotella,Gemella,Porphyromonas,Ruminococcaceae,Lachnospiraceae, Selenomonas, Clostridium, Haemophilus. The advantage bacterial colonies of HGS group: Lactococcus,Prevotella.Conclusion The flora structure of HFMD group was more complex, and there was obvious difference in the flora structure between HFMD group and HGS group.
Key words:  Polymerase chain reation-denaturing gradient gel electrophoresis(PCR-DGGE)  Hand, foot and mouth disease(HFMD)  Herpetic ginggivostomatitis(HGS)