Two novel PCR primers were designed and designated as UPTC flaF(forward primer) and UPTC flaR (reverse).
After PCR optimization, amplification was performed on all seven UPTC isolates, as well as with the C.
PCR amplification using the novel primer pair, UPTC flaF/ UPTC flaA, allowed the reproducible generation of a flaA amplicon of approximately the expected size (circa 1,358 bp) for all UPTC isolates examined (Fig.
After the original descriptions of UPTC appeared, isolates of UPTC were reported in France, Ireland and The Netherlands, and UPTC strains have also recently been found in Japan (Matsuda et al.
The understanding of sources and means of transmission of UPTC organisms is important to help elucidate the epidemiology of these organisms in shellfish and the marine environment, as well as their potential transmission to humans.
This study was interested in the ability to genotype members of the UPTC group of organisms isolated in shellfish and was undertaken to develop a PCR-based method that would allow the differentiation between UPTC organisms at the subspecies level.
After the development of a successful and reproducible PCR-RFLP system, employing the newly described primer pair (UPTC flaF/UPTC flaR), we wished to establish proof-of-principle for this technique in demonstrating the application of this method to a small collection of UPTC isolates obtained from the natural environment.