Diagnostic multiplex polymerase chain reaction assay for the identification of Pseudomonas aeruginosa from the skin biopsy specimens in burn wound infections and detection of antibiotic susceptibility

ABSTRACT

To identify Pseudomonas aeruginosa [P. aeruginosa] from the skin biopsy specimens in burn wound infections by multiplex polymerase chain reaction [M-PCR] and detection of antimicrobial susceptibility of isolates from culture. We conducted this cross-sectional study in 140 patients with wound infections who admitted to the referral burn center of Motahari, Tehran, Iran, during a 12-month period from 2005-2006. Skin biopsy specimens were aseptically taken from each patient, one for PCR and one for bacterial culture. A M-PCR test based on the simultaneous amplification of 2 lipoprotein genes oprI and oprL, was used to directly detect fluorescent pseudomonades and P. aeruginosa in skin biopsy specimens. The susceptibility of P. aeruginosa isolates to 16 antibiotics was determined using the disc diffusion method. Out of 140 biopsy specimens, M-PCR detected 66 [47.2%] isolates, while culture detected 57 [40.7%] isolates as P. aeruginosa. Positive results for both genes which observed only for P. aeruginosa, while only one gene, oprI, was amplified from other fluorescent pseudomonades n=12 and all other bacterial tested n=62 were negative by the amplification test. The most effective antibiotics against isolate of P. aeruginosa were cefepime [79%], azetreonam [76%], ticarcillin-clavulanic acid [68%], tobramycin [62%], and amikacin [61%]. Multiplex PCR assay appears promising for the rapid and sensitive detection of P. aeruginosa from the burned skin biopsy specimens. Simultaneous amplification of 2 lipoprotein genes oprI and oprL, could detect P. aeruginosa, and oprI gene only for other fluorescent pseudomonades