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

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Abstract:

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 g 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 Tn5281 amplicons. The number of HLGR isolates of E. faecalis has increased considerably in Tehran hospitals. Tn5281 is the main cause of transmission of aac(6′)-aph(2) to different isolates of E. faecalis in the hospitals studied.

Les infections par des isolats d’Enterococcus faecalis résistant à de hauts niveaux de gentamycine (HLGR) sont communes dans les hôpitaux de Téhéran. Les gènes responsables de cette résistance sont transmissibles par conjugaison à de hautes fréquences. Le but de cette étude était de détecter la présence du Tn5281 et du gène adjacent aac(6′)-aph(2″) codant une enzyme de modification des aminoglycosides, parmi les 102 isolats HLGR de E. faecalis cultivés à partir de patients de trois hôpitaux de Téhéran, Iran. Ces isolats ont été détectés sur des disques contenant 120 g de gentamycine et constituaient 65 % de tous les cas d’infection à E. faecalis à survenir pendant la période d’étude. L’ADN a été extrait des isolats HLGR et a été soumis à des essais PCR ciblant aac(6′)-aph(2″) et le transposon de conjugaison Tn5281. Le gène aac(6′)-aph(2″) amplifié a été marqué à la digoxigénine et hybridé aux amplicons Tn5281 par buvardage en point (dot blot). Le gène aac(6′)-aph(2″) a été détecté chez 91 % - 92 % (n = 93) des isolats HLGR. Tous les isolats contenant aac(6′)-aph(2″) étaient positifs en PCR longue ciblant le Tn5281 et la sonde s’hybridait avec les amplicons Tn5281. Le nombre d’isolats HLGR d’E. faecalis a considérablement augmenté dans les hôpitaux de Téhéran. Le Tn5281 est le principal responsable de la transmission de aac(6′)-aph(2″) aux différents isolats de E. faecalis dans les hôpitaux étudiés.

Document Type: Research Article

Publication date: October 1, 2008

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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