Specific detection of the toxic dinoflagellates Alexandrium tamarense and Alexandrium catenella from single vegetative cells by a loop-mediated isothermal amplification method.
In this study, we succeeded in developing a loop-mediated isothermal amplification (LAMP) method that enables sensitive and specific detection of the toxic marine dinoflagellates Alexandrium tamarense and Alexandrium catenella from single cells of both laboratory cultures and naturally blooming cells within 25 min, by monitoring the turbidimeter from the start of the LAMP reaction. The fluorescence intensity was strong enough to allow discrimination between positive and negative results by naked eye under a UV lamp, even in amplified samples from a single cell, by using the LAMP method. Unambiguous detection by naked eye was possible even in half the volume of LAMP cocktail recommended by the manufacturer, suggesting the potential to significantly reduce the cost of Alexandrium monitoring. Therefore, we can conclude that this method is one of the most convenient, sensitive, and cost-effective molecular tools for Alexandrium monitoring.
National Research Institute of Fisheries and Environment of Inland Sea, Maruishi 2-17-5, Hatsukaichi, Hiroshima 739-0452, Japan. snagai@affrc.go.jp
Satoshi
S
Nagai
Shigeru
S
Itakura
D000818:Animals; D017931:DNA Primers / Q000235:genetics; D004141:Dinoflagellida / Q000235:genetics* / Q000302:isolation & purification; D004784:Environmental Monitoring / Q000379:methods*; D005453:Fluorescence; D007564:Japan; D009391:Nephelometry and Turbidimetry / Q000379:methods; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000382:microbiology*
D017931:DNA Primers