In this study, we have developed a method for detecting Kudoa amamiensis using loop-mediated isothermal amplification (LAMP). After initial cloning and sequencing of the internal transcribed spacer (ITS) region between 18S and 28S rRNA genes of K. amamiensis, a set of four primers consisting of two inner and two outer was designed based on ITS1 and 5.8S rRNA sequence for use in the LAMP reaction. Reaction time and temperature were optimized for 60 min at 60°C, respectively. No cross-reaction of the LAMP method was observed with K. iwatai. Therefore, this LAMP technique was considered to be a rapid and simple detection method of K. amamiensis.
2015 The Japanese Society of Fish Pathology