Applying a PCR ‐based open‐reading frame typing method for easy genotyping and molecular epidemiological analysis of P seudomonas aeruginosa

Aims Molecular epidemiological techniques, such as pulsed‐field gel electrophoresis ( PFGE ), or multilocus sequence typing ( MLST ) have facilitated our understanding of the transmission routes of nosocomial infections by Pseudomonas aeruginosa . However, they are time consuming and technically demanding. To perform molecular epidemiological analysis in a standard microbiology laboratory, we aimed to develop a simpler and effective molecular epidemiological technique based on the open‐reading frame ( ORF ) distribution patterns detected by PCR , which we call PCR ‐based ORF typing ( POT ). Methods and Results Ten ORF s from genomic islets, five ORF s from genomic islands, and the metallo‐β‐lactamases ( MBL s) bla IMP and bla VIM were selected by comparing the whole‐genome sequences of different Ps. aeruginosa strains ( PAO 1, PA 7, UCBPP ‐ PA 14 and LESB 58). These 17 ORF s were detected, along with a Ps. aeruginosa marker, using 9‐plex and 10‐plex PCR systems. The genotypes in the POT were compared to those obtained by using PFGE and MLST . Conclusions Using the POT method, molecular epidemiological analyses of Ps. aeruginosa can be completed in 4 h. Significance and Impact of the Study Since this method is very easy to perform, even in standard clinical laboratories, it could be a valuable tool for monitoring daily infection control measures.