|Bassett, J - BACTERIAL BARCODES|
Submitted to: American Society for Microbiology
Publication Type: Abstract Only
Publication Acceptance Date: January 16, 2002
Publication Date: May 24, 2002
Citation: Siragusa, G.R., Hiett, K.L., Bailey, J.S., Craven, S.E., Stern, N.J., Bassett, J.J. 2002. Subtyping poultry-associated isolates of Campylobacter spp., Salmonella spp., and Clostridium perfringens by rep-PCR [Abstract]. American Society for Microbiology. p. 73. Technical Abstract: To gain enhanced genotypic resolution of bacterial strains from both common and different origins we have applied rep-PCR using a single primer set to three major food borne pathogens that are all poultry-associated or human clinical isolates taken within the same timeframe as the poultry isolates. The discrimination outcomes from rep-PCR using the primer Uprime Rdt (rep- PCRDt)were evaluated alongside other typing methods previously performed. Bacterial isolate libraries consisted of 50 isolates each of Campylobacter spp. and Cl. perfringens and 55 isolates representing six serotypes of salmonellae. Resulting dendograms were created by the UPGMA method. Duplicate patterns were highly reproducible. Based on visual dendogram analysis Campylobacter resolution from rep-PCRDt was very similar and in many cases identical to that based on flaA sequencing resolving into three pronounced clades (correlation >70%) along geographical lines and several smaller clades (correlation >90%). Within that library rep-PCRDt discriminated the only C. coli isolate from C. jejuni. Human and poultry C. jejuni isolates (collected in the same time frames from related sites) segregated into the same rep-PCRDt clade. Salmonella isolates resolved int rep-PCRDtgroups largely congruent to serotype; exceptions were within the Thompson and Typhimurium serogroups. Cl. perfringens data demonstrated processing plant isolates were found amongst clades from the breeder, hatchery and farm. Using standardized rep-PCRDt provided a means to effectively and reproducibly discriminate these pathogens that was straightforward, rapid and simple to perform. This method, used alone or in concert with other subtyping means enhances our ability to track the origin and relatedness of these bacteria.