Jansson J K, Prosser J I
Department of Biochemistry, Arrhenius Laboratories for Natural Sciences, Stockholm University, Sweden.
Mol Biotechnol. 1997 Apr;7(2):103-20. doi: 10.1007/BF02761746.
Traditional techniques for assessment of microbial numbers and activity generally lack the specificity required for risk assessment following environmental release of genetically engineered microbial inocula. Immunological and molecular-based techniques, such as DNA probing and genetic tagging, were initially used to determine the presence or absence of microorganisms in environmental samples. Increasingly they are being developed for quantification of populations of specific organisms, either indigenous or introduced, in the environment. In addition, they are being used to quantify the activity of particular organisms or groups of organisms, greatly extending the range of techniques available to the microbial ecologist. This article reviews the use of traditional techniques for the quantification of microbial population size and activity and the application of molecular techniques, including DNA probing, genetic marking, use of fluorescent probes, and quantitative PCR, in combination with advanced cell detection techniques such as confocal laser scanning microscopy and flow cytometry.
传统的微生物数量和活性评估技术通常缺乏对基因工程微生物接种物环境释放后进行风险评估所需的特异性。基于免疫和分子的技术，如DNA探针和基因标记，最初用于确定环境样品中微生物的存在与否。现在越来越多地将它们用于定量环境中特定生物体（无论是本土的还是引入的）的数量。此外，它们还被用于量化特定生物体或生物体群体的活性，极大地扩展了微生物生态学家可用的技术范围。本文综述了传统技术在微生物数量和活性定量方面的应用，以及分子技术的应用，包括DNA探针、基因标记、荧光探针的使用和定量PCR，这些技术与共聚焦激光扫描显微镜和流式细胞术等先进的细胞检测技术相结合。
