Sensitive and rapid detection of edwardsiellosis in fish by a loop-mediated isothermal amplification method.
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 degrees 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.
Savan
Ram
R
United Graduate School of Agricultural Sciences, Kagoshima University, Kagoshima 890-0065, Japan.
Igarashi
Arisa
A
Matsuoka
Satoru
S
Sakai
Masahiro
M
D000818:Animals; D017931:DNA Primers; D020609:Edwardsiella tarda / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004756:Enterobacteriaceae Infections / Q000382:microbiology / Q000662:veterinary*; D005393:Fish Diseases / Q000382:microbiology*; D005432:Flounder / Q000382:microbiology*; D006460:Hemolysin Proteins / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012623:Seawater / Q000382:microbiology; D012680:Sensitivity and Specificity; D013997:Time Factors
D017931:DNA Primers; D006460:Hemolysin Proteins