DGGE和qPCR联用定量分析环境样品中优势菌群  

Quantitative analysis of dominant bacteria in environmental samples based on DGGE plus quantitative PCR techniques

作　　者：付小花[1,3] 李艳丽[2] 乐毅全[1] 王磊[1] 

机构地区：[1]同济大学污染控制与资源化研究国家重点实验室,环境科学与工程学院,上海200092 [2]同济大学海洋地质国家重点实验室,上海200092 [3]上海市城市化生态过程与生态恢复重点实验室,上海200062

出　　处：《实验技术与管理》2012年第9期45-47,50,共4页Experimental Technology and Management

基　　金：国家自然科学基金资助项目(40571145,40871217);污染控制与资源化研究国家重点实验室基金资助项目(PCRRY11011);上海市城市化生态过程与生态恢复重点实验室开放基金资助(2010)

摘　　要：精确定量环境样品中特定微生物类群的数量对阐明环境生物治理/修复过程中微生物的功能和效应具有重要意义。该文以土壤样品为例,将PCR-DGGE技术与定量PCR(qPCR)技术联用,定量检测样品中优势微生物类群数量。方法:抽提样品DNA,针对16SrDNA进行PCR-DGGE分析,明确样品中的优势菌群;再根据特定优势菌群的基因序列设计适合的定量引物,通过针对特定优势菌群的定量PCR分析,得到样品中优势菌群的数量信息。该方法灵敏度高、重复性好,并且无需培养,最大限度地保持了样品原始群落信息。It is very important to precisely quantify the microbial groups in environmental samples for clarifying the functions and effects of microbes during the bio-treatment and bioremediation processes in degenerated environment. In this work, a method was developed to quantify the dominant microbial group through the DGGE plus quantitative PCR techniques. Firstly, the 16S rDNA was analyzed by PCR-DGGE after the total DNA was extracted from the environmental samples to identify the dominant bacteria secondly, primers could be designed according to the gene sequence of the dominant microbial groups, which could lead to quantitative results of dominant microbial groups after quantitative PCR analysis. The method demonstrates high sensitivity and ideal reproducibility in addition, it does not need a cultivating process, which could remain the original community information at the greatest degree.

关 键 词：优势菌群 定量检测 DGGE 定量PCR(qPCR) 

分 类 号：X172[环境科学与工程—环境科学]