| 基于LAMP-LFD技术的有害赤潮藻东海原甲藻(Prorocentrum donghaiense)快检方法 |
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| 引用本文： | 邹怡欣,黄海龙,乔龙亮,党晨阳,庞建虎,吕佩文,王佳棋,朱鹏.基于LAMP-LFD技术的有害赤潮藻东海原甲藻(Prorocentrum donghaiense)快检方法[J].海洋与湖沼,2018,49(3):586-593. |
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| 作者姓名： | 邹怡欣 黄海龙 乔龙亮 党晨阳 庞建虎 吕佩文 王佳棋 朱鹏 |
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| 作者单位： | 宁波大学海洋学院 |
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| 基金项目： | 宁波市科技攻关项目，2017C110003号；浙江省公益技术应用研究，2017C33133号；国家现代农业产业技术体系建设专项资金项目，CARS-47号。 |
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| 摘 要： | 以东海原甲藻的ITS序列(Internal transcribed spacer,ITS)为检测靶标,将生物素标记的环介导恒温扩增技术(Loop-mediated isothermal amplification,LAMP)扩增产物与经异硫氰酸荧光素(Fluorescein isothiocyanate,FITC)标记的探针特异性杂交,并结合横向流动试纸条(Lateral flow dipstick,LFD)肉眼直接观察检测结果,建立了有害赤潮藻东海原甲藻(Prorocentrum donghaiense)的快速检测技术。经优化后的最适条件为63°C、30min,较常规PCR扩增缩短约2h。结果表明:LAMPLFD可特异性检出东海原甲藻,对常见赤潮藻检测结果均为阴性;其对东海原甲藻基因组DNA的检测最低限为47pg/μL,是常规PCR技术(以F3/B3为引物)的10倍。LAMP-LFD技术能高效、特异地检出东海原甲藻,仪器设备依赖性低,结果可视化,有望成为赤潮原因种检测监控的常规方法。
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| 关 键 词： | 东海原甲藻 环介导恒温扩增技术 横向流动试纸条 生物快检 |
| 收稿时间： | 2017/8/18 0:00:00 |
| 修稿时间： | 2017/11/20 0:00:00 |
RAPID DETECTION OF PROROCENTRUM DONGHAIENSE BASED ON LOOP-MEDIATED ISOTHERMAL AMPLIFICATION COMBINED WITH LATERAL FLOW DIPSTICK |
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| ZOU Yi-Xin,HUANG Hai-Long,QIAO Long-Liang,DANG Chen-Yang,PANG Jian-Hu,L&#; Pei-Wen,WANG Jia-Qi and ZHU Peng.RAPID DETECTION OF PROROCENTRUM DONGHAIENSE BASED ON LOOP-MEDIATED ISOTHERMAL AMPLIFICATION COMBINED WITH LATERAL FLOW DIPSTICK[J].Oceanologia Et Limnologia Sinica,2018,49(3):586-593. |
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| Authors: | ZOU Yi-Xin HUANG Hai-Long QIAO Long-Liang DANG Chen-Yang PANG Jian-Hu L&#; Pei-Wen WANG Jia-Qi and ZHU Peng |
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| Institution: | Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China,Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China,Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China,Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China,Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China,Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China,Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China and Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China |
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| Abstract: | |
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| Keywords: | Prorocentrum donghaiense loop-mediated isothermal amplification (LAMP) lateral flow dipstick (LFD) quick detection |
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