山西医科大学学报2013，Vol.44Issue(7)：530-534,5.DOI:10.3969/J.ISSN.1007-6611.2013.07.007
16S rDNA克隆文库法与PCR-DGGE法在口腔菌群分析中的对比研究
Comparative study on 16S rDNA clone library versus PCR-DGGE method in analysis of oral bacterial flora
摘要
Abstract
Objective To compare the efficiency of 16S rDNA clone library versus PCR-DGGE method in detecting oral microbial diversity.Methods Bacterial samples were collected from root canals of 6 patients with retrograde pulpitis.Total DNA was extracted.The universal primers 27F/1492R and HAD-1/2 were used to amplify the full length or V2-V3 region of 16S rDNA,respectively.The PCR products were separated through agarose gel electrophoresis or denaturing gradient gel electrophoresis (DGGE).The bands were cloned and sequenced.The bacteria were identified by aligning the sequences in ribosomal database.Results One band of 1.5 kb corresponding to the full length of 16S rDNA was separated through agarose gel electrophoresis.DGGE fingerprints showed 6-12 bands of 239 bp corresponding to V2-V3 region of 16S rDNA.A total of 8-15 and 6-12 kinds of bacteria were detected in one sample by 16S rDNA and PCR-DGGE method,respectively.Conclusion Compared with PCR-DGGE method,16S rDNA clone library method is a more sensible and specific technology in detecting bacterial composition in the flora.关键词
口腔微生物群/16S rDNA/PCR-DGGE/克隆测序Key words
oral microbiota/ ribosomal 16S rDNA/ denaturing gradient gel electrophoresis(DGGE) / clone sequencing分类
医药卫生引用本文复制引用
赵焕英,尚佳健,张琛,关蕊,杨颖..16S rDNA克隆文库法与PCR-DGGE法在口腔菌群分析中的对比研究[J].山西医科大学学报,2013,44(7):530-534,5.基金项目
国家自然科学基金资助项目(81200783) （81200783）
北京市教育委员会科技计划面上基金资助项目(SQKM201210025019) （SQKM201210025019）
