应用RT_qPCR技术定量检测湖泊水体中蓝藻方法的比较.pdfVIP

微生物学通报 APR 20, 2011, 38(4): 460−467 Microbiology China © 2011 by Institute of Microbiology, CAS tongbao@ 研究报告 应用 RT-qPCR 技术定量检测湖泊水 体中蓝藻方法的比较 * 彭宇科 岳冬梅 武俊 肖琳 杨柳燕 ( 210046) 摘 要: 利用RT-qPCR 技术建立了对湖泊水体中的微囊藻和蓝藻的 SYBR Green I 荧光定量PCR 检测方法, 在所建立的方法中, 对以微囊藻藻蓝蛋白基因、蓝藻 16S rRNA 基因、微囊藻 16S rRNA 基因分别作为RT-qPCR 检测的目的基因所得结果进行了比较, 并对实验室培养的微囊藻和太湖的 环境样品进行了检测。结果表明, 用藻蓝蛋白基因作为检测目的基因, 以M. aeruginosa PCC 7806 基因组DNA 作为标准品的测定方法与显微镜计数的结果有较好的相关性和一致性, 并具有简便、 快速、特异性高的特点, 可以满足检测的要求。 关键词: 太湖, 蓝藻, 微囊藻, 实时荧光定量PCR, SYBR Green I Comparison of RT-qPCR approaches for quantification of cyanobacteria in Lake Taihu * PENG Yu-Ke YUE Dong-Mei WU Jun XIAO Lin YANG Liu-Yan (State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Nanjing University, Nanjing, Jiangsu 210046, China) Abstract: In the study, four methods used for quantification of cyanobacteria and Microcystis by SYBR Green I real-time quantitative PCR (RT-qPCR) were established based on the target genes, including the 16S rRNA gene of cyanobacteria, the phycocyanin and the 16S rRNA gene of Microcystis . Then the cultured M. aeruginosa and the environmental samples of Lake Taihu were evaluated by the established methods. Comparison between the new approaches and microscope counting indicated that R