Identification of commensal Pseudomonas aeruginosa isolates using duplex PCR targeting the oprL and algD genes

Background: Pseudomonas aeruginosa (P. aeruginosa) is a ubiquitous bacterium which can be found in most moisture places such as soil, water, food, plants, and animals including humans. Due to genetic flexibility among strains, there is no standard molecular identification for P. from different sources, particularly human colonizing or commensal isolates. Materials Method: Monoplex PCR using published primers of oprL, algD, nfxB gene were compared their specificity sensitivity aeruginosa. Twenty- nine isolates 16 13 aeruginosa-like used the study. To improve detection, two new primer pairs targeting oprL was designed (oprL-pp1 oprL-pp2), oprL-algD duplex carried out with newly pairs. Result conclusion: algD genes has same 93.75% 100%, while oprL-specific more sensitive (100%) but less specific (0%). Duplex yielded high detecting Both oprL-pp1/algD oprL-pp2/algD duplex-PCR had (15/16 isolates) 100% (0/13 isolates). Besides, oprL-pp2 than oprL-pp1 primers, only 2 over detected, detected all Compared monoplex that targeted gene, duplex-PCR utilizing identify 15/16 (93.75% specificity) reduce number oprL-positive required further exact identification. Additionally, this study negative non-Pseudomonas species E. coli, V. cholera, parahaemolyticus, S. aureus. In conclusion, our could valuable tool for P. aeruginosa screening.