|
|||||
|
|
| 检测肠道细菌Feacalibacterium prausnitzii的三对PCR引物的特异性比较 | |
| 引用本文： | 冯洁,华蔚颖,赵立平,赵宇峰,申剑.检测肠道细菌Feacalibacterium prausnitzii的三对PCR引物的特异性比较[J].微生物学报,2011,51(6):819-827. |
| 作者姓名： | 冯洁 华蔚颖 赵立平 赵宇峰 申剑 |
| 作者单位： | 1. 上海交通大学生命科学技术学院,教育部微生物代谢重点实验室,上海200240 2. 上海交通大学生命科学技术学院,教育部微生物代谢重点实验室;上海交通大学系统生物医学研究院,上海200240 3. 上海交通大学系统生物医学研究院,上海,200240 |
| 基金项目： | 国家自然科学基金重点项目（30730005） |
| 摘 要： | 【目的】比较3对基于16S rRNA基因、用于检测人肠道中重要细菌Feacalibacterium prausnitzii的引物(FPR-1/FPR-2、FPR-2F/Fprau645R和Fprau223F/Fprau420R)的特异性。【方法】用Clustal X比对每个引物与F.prausnitzii和其他细菌的16S rRNA基因的序列。在Ribosomal Database Project(RDP)数据库中使用Probe Match工具比较每个引物匹配的Faecalibacterium spp.序列数目。利用本课题组建立的中国人粪便菌群的16S rRNA基因全长文库的7255个克隆序列,用Simulated PCR(SPCR)预测每对引物检测到的F.prausnitzii和其他细菌的克隆数;用3对引物分别对代表克隆进行PCR扩增。用3对引物分别对14个健康人的粪便样品进行实时定量PCR。【结果】引物Fprau645R的3端最后一个碱基与非F.prausnitzii序列的错配度高于其它引物,它在RDP中匹配的Faecalibacterium spp.序列数占其匹配的细菌序列数的百分比(97.6%)显著高于其他引物。SPCR预测,3对引物检测到的F.prausnitzii克隆数均为1171左右;在检测到的非Faecalibacterium spp.克隆中,FPR-2F/Fprau645R主要是Subdoligranulum spp.,而FPR-1/FPR-2和Fprau223F/Fprau420R还有Oscillibacter spp.、Ruminococcus spp.和unclassified Ruminococcaceae等。真实PCR与SPCR的结果吻合。实时定量PCR中,FPR-1/FPR-2和Fprau223F/Fprau420R检测到的细菌数量高于FPR-2F/Fprau645R。【结论】3对引物能检测到F.prausnitzii和Subdoligranulum spp.,FPR-2F/Fprau645R的特异性优于FPR-1/FPR-2和Fprau223F/Fprau420R。 |
| 关 键 词： | Feacalibacterium prausnitzii 引物 特异性 肠道菌群 |
| 收稿时间： | 2011/1/21 0:00:00 |
| 修稿时间： | 3/6/2011 12:00:00 AM |
Specificity comparison of three Feacalibacterium prausnitzii-specific PCR primer pairs |
|
| Jie Feng,Weiying Hu,Liping Zhao,Yufeng Zhao and Jian Shen.Specificity comparison of three Feacalibacterium prausnitzii-specific PCR primer pairs[J].Acta Microbiologica Sinica,2011,51(6):819-827. | |
| Authors: | Jie Feng Weiying Hu Liping Zhao Yufeng Zhao and Jian Shen |
| Affiliation: | Key Laboratory of Microbial Metabolism, Ministry of Education, College of Life Science and Biotechnology,Shanghai 200240, China;Key Laboratory of Microbial Metabolism, Ministry of Education, College of Life Science and Biotechnology,Shanghai 200240, China;Key Laboratory of Microbial Metabolism, Ministry of Education, College of Life Science and Biotechnology,Shanghai 200240, China;College of Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, China;College of Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, China |
| Abstract: | Objective] To compare the specificity of three 16S rRNA gene-based PCR primer pairs(FPR-1/FPR-2,FPR-2F/Fprau645R and Fprau223F/Fprau420R),which are used to specifically detect and quantify the important human gut bacterium Feacalibacterium prausnitzii.Methods] Clustal X was used to align the sequences of individual primer and the 16S rRNA gene of F.prausnitzii and other bacteria.The number of Faecalibacterium spp.sequences in Ribosomal Database Project(RDP) matched by each primer was obtained by the Probe... |
| Keywords: | Keywords: Feacalibacterium prausnitzii primer specificity gut microbiota |
| 本文献已被 CNKI 万方数据 PubMed 等数据库收录！ | |
| 点击此处可从《微生物学报》浏览原始摘要信息 | |
| 点击此处可从《微生物学报》下载全文 | |
