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Applied and Environmental Microbiology, January 2004, p. 621-624, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.621-624.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Sensitive and Rapid Detection of Edwardsiellosis in Fish by a Loop-Mediated Isothermal Amplification Method

Ram Savan,1 Arisa Igarashi,1 Satoru Matsuoka,2 and Masahiro Sakai3*

United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima 890-0065,1 Ehime Prefectural Chuyo Fisheries Experimental Station, Iyo City, Ehime 799-3125,2 Faculty of Agriculture, Miyazaki University, Miyazaki 889-2192, Japan3

Received 7 July 2003/ Accepted 7 October 2003

Here we report a rapid and sensitive method (using loop-mediated isothermal amplification [LAMP]) for the diagnosis of edwardsiellosis, a fish disease caused by Edwardsiella tarda, in Japanese flounder. A set of four primers was designed, and conditions for the detection were optimized for the detection of E. tarda in 45 min at 65°C. No amplification of the target hemolysin gene was detected in other related bacteria. When the LAMP primers were used, detection of edwardsiellosis in infected Japanese flounder kidney, and spleen and seawater cultures was possible. We have developed a rapid and sensitive diagnostic protocol for edwardsiellosis detection in fish. This is the first report of the application of LAMP for the diagnosis of a fish pathogen.

* Corresponding author. Mailing address: Masahiro Sakai Faculty of Agriculture, Miyazaki University, Gakuen kibanadai nishi 1-1, Miyazaki 889-2192, Japan. Phone and fax: 81-985-587219. E-mail: m.sakai{at}cc.miyazaki-u.ac.jp.

Applied and Environmental Microbiology, January 2004, p. 621-624, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.621-624.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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