产毒微囊藻PCR检测方法的建立  

Development of PCR Method for Detection of Potential Microcystin-producing Microcystis

作　　者：周毅[1] 徐晔[1] 钱伟[2] 姚燕林[1] 王海涛[1] 张敬友[3] 王维志[1] 段宏安[1] 

机构地区：[1]连云港出入境检验检疫局,江苏连云港222042 [2]苏州出入境检验检疫局,江苏苏州215000 [3]南京出入境检验检疫局,江苏南京210001

出　　处：《安徽农业科学》2013年第1期43-45,共3页Journal of Anhui Agricultural Sciences

基　　金：质检公益项目(201110037);质检总局科技项目(2010-IK005)

摘　　要：[目的]建立产毒微囊藻的PCR检测方法。[方法]根据GenBank上发表的mcyA基因序列保守区域设计一对引物,建立和优化检测产毒微囊藻的PCR方法,并以此方法检测产毒和非产毒微囊藻参考株系。[结果]经PCR检测,产毒藻株出现特异性扩增条带,而不产毒株未出现特异性条带;以10倍系列稀释纯培养的蓝藻细胞作为PCR反应的模板,检测限均为2.46×103细胞/ml;运用建立的方法从天然水体中检测到了产毒微囊藻,对PCR产物序列进行比对,发现PCR扩增片段与铜绿微囊藻(PCC7806)核苷酸序列的同源性为98%。[结论]该研究为水华产毒微囊藻的监测提供了一种新方法。[ Objective ] The aim was to develop a PCR method for detection of potential microcystin-producing microcystis. [ Method ] Accord- ing to conserved regions of mcyA gene sequences in GenBank, a pair of primers was designed, and a PCR method for the detection of microcys- tin-producing microcystis was developed and optimized. Then the referenced microcystin-producing and non-microcystin-producing microcystis strains were detected by this method. [ Result] The microcystin-producing microcystis stains appeared specific DNA band and the non-micro- cystin-producing microcystis strains didn＇t show specific band by the PCR test; the intact cells were 10 times diluted and used as template, and the results indicated that the detection limit of the method was up to 2.46 × 103 cells/ml ; the microcystin-producing microcystis was de- tected from natural water by using the established method and the Blast N result showed that this PCR fragments had 98% homology to that of Microcystis aeruginosa （PCC7806）. [ Conclusion ] This study provided a new method for the monitoring of microcystin-producing microcystis in bloom.

关 键 词：产毒微囊藻 mcyA PCR 检测 同源性 

分 类 号：S188[农业科学—农业基础科学]