Sensitive and rapid detection of two toxic microalgae Alexandrium by loop-mediated isothermal amplification |
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| Authors: | ZHANG Fengying SHI Yanhong JIANG Keji XU Zhaoli and MA Lingbo |
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| Institution: | 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 |
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| Abstract: | A loop-mediated isothermal amplification (LAMP) assay was designed and evaluated for rapid de- tection 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 65ffiC. 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 lim- its 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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| Keywords: | Alexandrium catenella Alexandrium minutum detection loop-mediated isothermal amplification (LAMP) ribosomal DNA internal transcribed spacer (ITS) |
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