应用RT-qPCR技术定量检测湖泊水体中蓝藻方法的比较

Comparison of RT-qPCR approaches for quantification of cyanobacteria in Lake Taihu

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

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 RT-qPCR analysis could meet the requirement of algae bloom quantification and it was convenient, rapid and specific.

Key words:Lake Taihu, Cyanobacteria, Microcystis, real-time quantitative PCR, SYBR Green I