Xia L.,Guangdong Ocean University |
Xia L.,Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals |
Xia L.,Key Laboratory of Diseases Controlling for Aquatic Economic Animals of Guangdong Higher Education Institutions |
Zhang H.,Guangdong Ocean University |
And 12 more authors.
Journal of Microbiology and Biotechnology | Year: 2015
Nocardia salmonicida is one of the main pathogens of fish nocardiosis. The purpose of this study was to build a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of N. salmonicida. A set of four primers were designed from the 16S-23S rRNA intergenic spacer region of N. salmonicida, and conditions for LAMP were optimized as incubating all the reagents for 60 min at 64°C. LAMP products were judged with agar gel electrophoresis as well as with the naked eye after the addition of SYBR Green I. Results showed the sensitivity of the LAMP assay was 1.68 × 103 CFU/ml (16.8 CFU per reaction) and 10-fold higher than that of PCR. The LAMP method was also effectively applied to detect N. salmonicida in diseased fish samples, and it may potentially facilitate the surveillance and early diagnosis of fish nocardiosis. © 2015 by The Korean Society for Microbiology and Biotechnology.