Transposon Tn5281 is the main distributor of the aminoglycoside modifying enzyme gene among isolates of Enterococcus faecalis in Tehran hospitals

Infections with high levels of gentamicin-resistant (HLGR) isolates of Enterococcus faecalis are common in Tehran hospitals. Genes encoding such resistance are transmissible by conjugation at high frequency. The purpose of this study was to determine the existence of Tn5281 and its flanking aminoglycoside modifying enzyme gene aac(6′)-aph(2″) among 102 HLGR isolates of E. faecalis cultured from patients at three hospitals in Tehran, Iran. These isolates were detected by disks containing 120 μg of gentamicin and made 65% of all E. faecalis during the study period. DNA was extracted from HLGR isolates and subjected to PCR assays targeting aac(6′)-aph(2″) and conjugative transposon Tn5281. The amplified aac(6′)-aph(2″) gene was labeled with digoxigenin and probed with Tn5281 amplicons in dot blot hybridization assays. The aac(6′)-aph(2″) gene was detected in 91%–92% (n = 93) of the HLGR isolates. All isolates containing aac(6′)-aph(2″) were positive in long-PCR targeting Tn5281 and the probe hybridized with ...