Melissococcus plutonius is the causative agent of European foulbrood (EFB) of honeybees. On the basis of phenotypic characteristics, M. plutonius strains are classified into those with typical (typical M. plutonius) and atypical characteristics (atypical M. plutonius), and the two types were suggested to have different pathogenic mechanisms. Therefore, differentiation of typical and atypical M. plutonius is important for more effective control of EFB. In this study, we developed a duplex PCR assay to detect and differentiate typical and atypical M. plutonius rapidly and easily. In this PCR, Na+/H+ antiporter gene and Fur family transcriptional regulator gene were selected as targets for typical and atypical strains, respectively. Under optimized conditions, this PCR system successfully detected M. plutonius from 50 copies of chromosome and differentiated between typical and atypical strains. No amplification was seen in any of the approximately 30 other bacterial species tested, indicating specificity of this method. Moreover, the duplex PCR can detect typical and atypical M. plutonius directly from diseased honeybee larvae. These results suggest the usefulness of this PCR assay for rapid diagnosis and epidemiological study of EFB.
(Bacterial and Parasitic Diseases Research Division)
References:
Arai R. et al. (2014) J. Vet. Med. Sci. 76:491-498
