For the rapid identification of , multiplex PCR, using 16S rDNA as target gene, was used and its value was evaluated. Forward primers specific for and , among the five validly described species, were designed from the alignment of 16S rDNA sequences. has been shown to be a member of the same species as by recent molecular studies and preliminary DNA-DNA relatedness tests. Therefore, and were considered as members of the same species in this study. Each primer was found to be species-specific by specificity testing. NSP01, NSP19, NSP21and NSP22could be clearly differentiated by PCR products characteristic for each species in the multiplex PCR assay. gave an identical result to NSP01. The additional 17 strains of and the additional four strains of gave PCR products identical to those of NSP01and NSP22, respectively. Multiplex PCR was found to be rapid, species-specific and reproducible. The technique evaluated in this study proved to be effective for rapidly identifying strains to species level.


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