Original articles Identification of PSE and OXA β-lactamase genes in Pseudomonas aeruginosa using PCR-restriction fragment length polymorphism

Methods: Two hundred and fifty-nine P. aeruginosa isolates were screened by PCR with 11 primer pairs designed to detect genes encoding PSE, OXA ,T EMand SHV enzymes. PSE and OX Ag ene variants were distinguished by restriction of PCR products with endonucleases recognizing sites involved in point mutations .N ucleotide sequences were verified for a few isolates by sequencing the PCR products. Results: Four isolates produced extended-spectrum β β β β-lactamases (ESBLs) according to the double disc synergy test. PCR detecting blaPSE genes was positive in 162 (62.5%) isolates: 151 carried blaPSE-1 and 11 carried a variant encoding an enzyme differing from PSE-1 by a single amino acid substitution (Pro102 to Ser). PCR detecting sequences for enzymes of the OXA-10 group was positive in 68 (26.3%) isolates: 31 carried blaOXA-10, one carried blaOXA-14 and 36 carried a new variant intermediate between blaOXA-13 and blaOXA-19 .T he blaOXA-2 gene was identified in 13 (5%) isolates .T wo other isolates carried blaOXA-2 variants encoding ESBLs differing from OXA-2 by a single amino acid substitution (Asp150 to Tyr and Trp159 to Cys, respectively). PCR detecting sequences for enzymes of the OXA-1 group was positive in 12 (4.6%) isolates. A blaTEM gene was identified in five (1.9%) isolates (three blaTEM-1, one blaTEM-2, one blaTEM-4). Conclusion: This approach is effective for screening P. aeruginosa for β β β β-lactamase gene