基于mcyJ基因序列的酶联夹心杂交法检测产毒微囊藻

Detection of Microcystin-producing Microcystis Based on Gene mcyJ

微囊藻是分布最广的一类水华蓝藻,多数都能产生微囊藻毒素。研究、建立产毒微囊藻的快速检测方法具有十分重要的意义。针对藻毒素合成基因mcyJ设计一组可用于产毒微囊藻种属的特异性探针,以实验室培养的微囊藻FACHB 905为研究对象,建立了一种检测产毒微囊藻的酶联夹心杂交法,并对酶联夹心杂交实验中的杂交液成分、浓度,杂交时间、温度以及酶联反应时间和温度等反应条件逐一进行了优化,建立了该方法的标准曲线,相关系数为0.997。

Cyanobacteria of the genus Microcystis are among the most common bloom-forming species in lakes and reservoirs worldwide.Most of them can produce microcystins.It is important to establish a rapid detection method for microcystin-producing Microcystis.Based on sequence analyses of microcystin-synthesis gene mcyJ from microcystin-producing Microcysti strains,a set of genus-specific probe pair was designed,and a sandwich hybridization assay was established using Microcystis FACHB 905 incubated in the laboratory as reseach objective.Experimental parameters,such as hybridization solution components and concentration,hybridization time and temperature as well as enzyme-linked reaction time and temperature were optimized.The standard curve was established,and correlation coefficient was 0.997.