To investigate the presence of Fusobacterium necrophorum (F. necrophorum) and Dichelobacter nodosus (D. nodosus) in dairy cattle footrot, a loop-mediated isothermal amplification method for detection of D. nodosus (Dn-LAMP) was developed and evaluated using species-specific16S rRNA gene as target gene. The Dn-LAMP showed no cross reaction with several common pathogens from cattle, and detection limit of the Dn-LAMP was 10 cfu/mL of D. nodosus. Amplicons of the Dn-LAMP could be detected by visual inspection. 189 hoof swabs from footrot-infected dairy cows in Heilongjiang province of China were detected using Dn-LAMP and Fn-LAMP for diagnosis of F. necrophorum, respectively. In 189 hoof swabs, 9 were positive for D. nodosus, and 95 were positive forF. necrophorum. These data suggested that combination of Dn-LAMP with Fn-LAMP will have a potential use for etiology investigation of footrot in dairy cattle.
Key words: Dichelobacter nodosus, Fusobacterium necrophorum, LAMP.
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