| ZHANG Fengying,SHI Yanhong,JIANG Keji,XU Zhaoli,MA Lingbo. 2012. Sensitive and rapid detection of two toxic microalgae Alexandrium by loop-mediated isothermal amplification. Acta Oceanologica Sinica, (2):139－146 |
| Sensitive and rapid detection of two toxic microalgae Alexandrium by loop-mediated isothermal amplification |
| Sensitive and rapid detection of two toxic microalgae Alexandrium by loop-mediated isothermal amplification |
| Received:December 20, 2010 Revised:August 08, 2011 |
| DOI：10.1007/s13131-012-0200-5 |
| Key words:Alexandrium catenella Alexandrium minutum detection loop-mediated isothermal amplification (LAMP) ribosomal DNA internal transcribed spacer (ITS) |
| 中文关键词: Alexandrium catenella Alexandrium minutum detection loop-mediated isothermal amplification (LAMP) ribosomal DNA internal transcribed spacer (ITS) |
| 基金项目:The Science and Technology Commission of Shanghai Municipality under contract Nos 062358101, 08DZ1980802 and 10JC1418600; a special research fund for the national non-profit institutes (East China Sea Fisheries Research Institute) under contract Nos 2007M22 and 2007Z01. |
| Author Name | Affiliation | E-mail | | ZHANG Fengying | East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, China
Key and Open Laboratory of Marine and Estuarine Fisheries, Ministry of Agriculture of China, Shanghai 200090, China | | | SHI Yanhong | East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, China
Key and Open Laboratory of Marine and Estuarine Fisheries, Ministry of Agriculture of China, Shanghai 200090, China | | | JIANG Keji | East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, China
Key and Open Laboratory of Marine and Estuarine Fisheries, Ministry of Agriculture of China, Shanghai 200090, China | | | XU Zhaoli | East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, China
Key and Open Laboratory of Marine and Estuarine Fisheries, Ministry of Agriculture of China, Shanghai 200090, China | | | MA Lingbo | East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, China
Key and Open Laboratory of Marine and Estuarine Fisheries, Ministry of Agriculture of China, Shanghai 200090, China | malingbo@sina.com |
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| Abstract: |
| A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid detection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning (PSP). Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer (ITS) of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection limits of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction (PCR) method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum. |
| 中文摘要: |
| A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid detection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning (PSP). Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer (ITS) of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection limits of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction (PCR) method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum. |
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