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Abstract

Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid diagnostic method that can rapidly amplify a target template under isothermal conditions. Using the LAMP method, a highly specific and sensitive diagnostic system for grouper iridovirus (GIV) detection was designed. Two outer and two inner primers were designed from GIV genome DNA. Reaction time and temperature were optimized for 60 min at 65°C, respectively. The detection limit of LAMP was about 100 ag and 100 times more sensitive than polymerase chain reaction. LAMP was carried out under various heat source reactor without any particular equipment and the reaction products are observed directly, after the addition of ethidium bromide can enhance the inspection of naked-eye. A diagnostic procedure which is rapid, highly sensitive and specific was developed for GIV detection

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