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Fish Pathology
Vol. 50 (2015) No. 3 p. 119-122

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http://doi.org/10.3147/jsfp.50.119

Short Communications

Detection of Kudoa amamiensis Using Loop-Mediated Isothermal Amplification (LAMP)

Masahiro Sakai1), Gouranga Biswas2), Tomoya Kono1), Jun-ichi Hikima1), Hiroshi Yokoyama3)

1) Department of Biochemistry and Applied Biosciences, Faculty of Agriculture, University of Miyazaki 2) Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki 3) Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo

Released on J-STAGE 20150929  
Received 20150313  

Keywords: Kudoa amamiensis, ITS region, molecular diagnosis, loop-mediated isothermal amplification, LAMP


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.

Copyright © 2015 The Japanese Society of Fish Pathology
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