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Background  Enteric Redmouth (ERM) disease also known as Yersiniosis is a contagious disease affecting salmonids, mainly rainbow trout. The causative agent is the gram-negative bacterium  Yersinia ruckeri . The disease can be diagnosed by isolation and identification of the causative agent, or detection of the  Pathogen  using fluorescent antibody tests, ELISA and PCR assays. These diagnostic methods are laborious, time consuming and need well trained personnel.    Results  A loop-mediated isothermal amplification (LAMP) assay was developed and evaluated for detection of  Y. ruckeri  the etiological agent of enteric red mouth (ERM) disease in salmonids. The assay was optimised to amplify the  yruI/yruR  gene, which encodes  Y. ruckeri  quorum sensing system, in the presence of a specific primer set and  Bst  DNA polymerase at an isothermal temperature of 63°C for one hour. Amplification products were detected by visual inspection, agarose gel electrophoresis and by real-time monitoring of turbidity resulted by formation of LAMP amplicons. Digestion with  Hph I restriction enzyme demonstrated that the amplified product was unique. The specificity of the assay was verified by the absence of amplification products when tested against related bacteria. The assay had 10-fold higher sensitivity compared with conventional PCR and successfully detected  Y. ruckeri  not only in pure bacterial culture but also in tissue homogenates of infected fish.    Conclusion  The ERM-LAMP assay represents a practical alternative to the microbiological approach for rapid, sensitive and specific detection of  Y. ruckeri  in fish farms. The assay is carried out in one hour and needs only a heating block or water bath as laboratory furniture. The advantages of the ERM-LAMP assay make it a promising tool for molecular detection of enteric red mouth disease in fish farms.

bmc veterinary research

Loop-mediated isothermal amplification as an emerging technology for detection of Yersinia ruckerithe causative agent of enteric red mouth disease in fish