实时荧光定量PCR及其在微生物生态学中的应用  

Real-time fluorescent quantitative PCR and its application in microbial ecology

作　　者：张晶[1] 张惠文[1] 张成刚[1] 

机构地区：[1]中国科学院沈阳应用生态研究所

出　　处：《生态学报》2005年第6期1445-1450,共6页Acta Ecologica Sinica

基　　金：中国科学院知识创新工程前沿领域探索资助项目(C12MGSCX-MS-0107)~~

摘　　要：定量描述微生物群落的组成,在微生物生态学的许多研究领域都是非常重要的。然而由于可培养技术的局限性,定量描述微生物群落成为比较困难的事情。最近包括PCR技术在内的分子生物学技术为人们提供了有力的工具,使对微生物群落的分布、丰度等有了进一步的了解。实时荧光定量PCR技术作为核酸定量检测技术,自从发明以来在微生物生态学研究中逐渐得到了广泛的应用。从微生物生态学角度,综述了实时荧光定量PCR技术的原理、发展、优缺点及其在微生物生态学研究中的应用与研究进展,并探讨了实时荧光定量PCR技术的发展和应用前景。The quantitative description of microbial community, especially the dominant groups and the groups that play some biological functions, is very important in some aspects of microbial ecology. However, conventional cultivation techniques cannot be used to fully characterize most soil microorganisms in respect that most of environmental species cannot be cultured, so it brings us some obstacles and makes us not thoroughly study the microbial communities. Fortunately, some molecular biological techniques including PCR technique give us new ways to understand the distribution and abundance of microbial community. Since the invention of real-time fluorescent quantitative PCR as a quantitative technique of nucleic acid, due to its high sensitivity and high accuracy, it has been more and more wildly applied in the field of microbial ecology. Using real-time fluorescent quantitative PCR profile to characterize microbial community can overcome the limitation of traditional technique, moreover accurate information about the microbial community can be got. The development of real-time fluorescent quantitative PCR, its principle and advantages and disadvantages were described in detail, furthermore the application and advances of real-time PCR in this field were reviewed. Future trends of real-time PCR techniques in microbial ecology were also discussed. Real-time PCR is a powerful tool for analysis of microbial communities. In order to get the more accurate and full information, it is better to combine with other molecular and microbiological techniques to give us a more clearly detailed view of microbial community structure, composition and abundance. At the same time, how to overcome some factors that affect the result of real-time PCR is another challenge we have to affront with. With the development of real-time PCR technique, the study strives for realizing absolute quantification and universal quantification in the future.

关 键 词：实时荧光定量PCR技术 SYBR Green I 荧光探针 微生物生态学 

分 类 号：Q938.1[生物学—微生物学]