Novel PCR-RFLP system based on rpoB gene for differentiation of Cronobacter species

• Jiří Vlach ^[1] ; Barbora Javůrková ^[1] ; Ludmila Karamonová ^[1] ; Martina Blažková ^[1] ; Ladislav Fukal ^[1]
  1. [1] University of Chemistry and Technology

     University of Chemistry and Technology

     Chequia

     
• Localización: Food microbiology, ISSN 0740-0020, Vol. 62 (April), 2017, págs. 1-8
• Idioma: inglés
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• Resumen

  • Bacteria from the genus Cronobacter are opportunistic foodborne pathogens that can cause severe infections. More rapid, cost-effective and reliable methods are still required for the species identification of Cronobacter spp. In this study, we present a novel PCR-RFLP-based method that uses a newly designed pair of primers for the PCR-amplification of a partial rpoB gene sequence (1635 bp). The amplified products of DNA from 80 Cronobacter strains were separately digested with three restriction endonucleases (Csp6I, HinP1I, MboI). Using the obtained restriction patterns, a PCR-RFLP identification system was created to enable differentiation between all seven currently-known Cronobacter species. The functionality of our method was successfully verified on real food samples. Moreover, the relationships between the Cronobacter species were determined via a phylogenetic tree created from the RFLP patterns.