产毒微囊藻PCR检测方法的建立 |
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引用本文: | 周毅,徐晔,钱伟,姚燕林,王海涛,张敬友,王维志,段宏安. 产毒微囊藻PCR检测方法的建立[J]. 安徽农业科学, 2013, 41(1): 43-45 |
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作者姓名: | 周毅 徐晔 钱伟 姚燕林 王海涛 张敬友 王维志 段宏安 |
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作者单位: | 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 |
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基金项目: | 质检公益项目,质检总局科技项目 |
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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。
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关 键 词: | 产毒微囊藻 mcyA PCR 检测 同源性 |
Development of PCR Method for Detection of Potential Microcystin-producing Microcystis |
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Affiliation: | ZHOU Yi et al(Lianyungang Entry-exit Inspection & Quarantine Bureau,Lianyungang,Jiangsu 222042) |
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Abstract: | [ 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. |
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Keywords: | Toxigenic microcystis mcyA PCR Detection Homology |
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