Prevalence of aac(6')-Ie-aph(2″)-Ia resistance gene and its linkage to Tn5281 in Enterococcus faecalis and Enterococcus faecium isolates from Tabriz hospitals
Abstract
Background and Objective: High-level gentamicin resistance (HLGR: MIC ≥ 500 µg/ml) in Enterococci is mediated by aminoglycoside modifying enzymes which is mainly encoded by aac(6′)-Ie-aph(2′′)-Ia gene. The aim of this study was to evaluate the frequency of aac(6′)-Ie-aph(2′′)-Ia gene in clinical isolates of Enterococcus facium and Enterococcus faecalis collected from hospitals in northwest of Iran.
Materials and methods: In the present study a total of 111 enterococcus isolates were collected from 4 hospitals during a two year period (July 2009-August 2011). Bacterial identification and species determination were carried out by standard biochemical tests. Antimicrobial susceptibility was evaluated by Kirby Bauer disc diffusion method. MICs were determined by agar dilution method. The frequency of aac(6′)Ie-aph(2″)Ia gene in the isolates was determined by PCR. The carriage of resistance gene on Tn5281 transposon was identified by long PCR and dot-blot hybridization methods.
Results: Antibiotic susceptibility tests revealed that the highest resistance was against streptomycin (74.77%) and erythromycin (67.58%) whereas the highest susceptibility was observed to vancomycin (81.1%). 36 isolates (32.43%) were identified as HLGR, 34(94.44%) of them had resistant gene in their genome. Long PCR studies revealed that 88 % of HLGR clinical isolates harboured Tn5281. The aac(6′)Ie-aph(2″)Ia resistance gene was present on Tn5281 transposon in all 32 isolates according to dot blot hybridization test.
Conclusion: The results of this study indicated that aac(6′)Ie-aph(2″)Ia resistance gene is highly prevalent in gentamicin resistant isolates. Carriage of aac(6′)Ie-aph(2″)Ia resistance gene on Tn5281 transposable element suggests possible contribution of this transposone on dissemination of resistance gene among enterococcus isolates.
Karlowsky JA, Jones ME, Draghi DC, Thorns Berry C, Sahm DF, Volturo GA, et al. Prevalence and antimicrobial susceptibilities of bacteria isolated from blood cultures of hospitalized patients in the United States in 2002. Ann Clin Microbiol Antimicrob 2004;10: 3-7.
Maki DG, Agger WA. Enterococcal bacteremia: clinical features, the risk of endocarditis, and management. Medicine (Baltimore) 1988; 67: 248-269.
Murray BE. The life and times of the Enterococcus. Clin Microbiol Rev. 1990; 3: 46-65.
Horodniceanu T. Bougueleret L, El-Solh N, Bieth G, Delbos F. High level plasmid-borne resistance to gentamicin in Streptococcus faecalis subsp. Zymogenes. Antimicrob Agents Chemother 2007; 16: 686-689.
Mayer, KH, Opal SM, Medeiros AA (2000). Mechanisms of antibiotic resistance. In: Principles and Practice of Infectious Diseases. Eds, Man dell , G. L., Bennett, J. E.
& Dolin, R. 5th ed. Churchill Livingstone, New York. pp.212-25.
Fisher K, Phillips C. The ecology, epidemiology and virulence of Enterococcus. Microbiology 2009; 155:1749-1757.
Sheppard BD, Gilmore MS. Antibiotic-resistant enterococci: the mechanisms and dynamics of drug introduction and resistance. Microbes Infect 2002; 4:215-224.
Thal LA, Chow JW, Patterson J. Molecular characterization of highly gentamicin-resistant enterococcus faecalis isolates lacking high-level streptomycin resistance. Antimicrob Agents Chemother 1993; 37: 134-137.
Huycke MM, Sahm DF, Gilmore MS. Multiple drug resistant enterococci: the nature of the problem and an agenda for the future. Emerg Infect Dis 1998; 4: 239-249.
Woodford N, Morrison D, Cookson B, George RC. Comparison of high-level gentamicin resis- tant Enterococcus faecium isolates from different continents. Antimicrob Agents Chemother 1993; 37:681-684.
Swenson JM, Ferraro MJ, Sahm DF, Clark NC, Culver DH. Tenover FC. Multilaboratory evaluation of screen- ing methods for detection of high-level aminoglycoside resistance in enterococci. J Clin Microbiol 1995; 33:3008-3018.
Blanch AR. Identification of Enterococcus spp. with a biochemical key. Appl Environ Microbiol 1999; 65:4425-4430.
Clinical and laboratory standards institute (CLSI).Methods for dilution antimicrobial susceptibility tests for bacteria that grow. aerobically: Approved Standard 8th Edition. CLSI document M07-A8 (ISBN 1-56238-689-1). Clinical and laboratory standards institute, USA, 2009.
Regnault B. Universal ribotyping methods using a chemically labelled oligonucleotide probe mixture. Res Microbiol 1997; 148: 649-659.
van de Klundert JAM, Vliegenthart JS. PCR detection of genes coding for amino glycoside-modifying enzymes. In : Diagnostic Molecular Microbiology: Principles and Applications . Eds , Persing, D. H., Smith, T. F., Tenover, F.C. & White, T. J. American Society for Microbiology, Washington, DC. 1993. pp. 547-552.
Woodford N, Mcnamara E, Smyth E, George RC. High- level resistance to gentamicin in enterococcus faecium. J Antimicrob Chemother1992; 29: 395-403.
Expand long template PCR system. Deoxynucleoside- triphosphate:DNA deoxy nucleotidyl transferase (2005), E.C.2.7.7.7. Roche diagnostics, GmbH, Mannheim, Germany.
Zarrilli R, Tripodi MF, Fortunato R, Bagattini M.Molecular epidemiology of high-level aminoglycoside- resistant enterococci isolated from patients in a university hospital in southern Italy. J Antimicrob Chemother 2005; 56: 827-835.
Arvanitidou M, Katsouyannopoulos V, Tsakris A.Antibiotic resistance patterns of enterococci isolated from coastal bathing waters. J Med Microbiol 2001; 501001-1005.
Simjee S, Gill MJ. Gene transfer, gentamicin resistance and enterococci. J Hosp Infect 1997; 36: 249-259.
Mrenda G, Lee L, Kelly C, Solorzano F, Leaños B, Muñoz O, et al. Antimi crobial resistance from enterococci in a pediatric hospital. Plasmids in Enterococcus faecalis isolates with high level gentamicin and streptomycin resistance. Arch Med Res. 2001: 32: 159-163.
Billstrom H, Lund B, Sullivan A, Nord CE. Virulence and antimicrobial resistance in clinical Enterococcus faecium. Int J Antimicrob Agents 2008; 32: 374-377.
Patterson JE, Masecar BL, Kauffman CA, Schaberg DR, Hierholzer WJ Zervos MJ. Gentamicin resistance plasmids of enterococci from diverse geographic areas are heterogeneous. J Infect Dis 1988; 158: 212-216.24. Dadfarma N, Oskouei M, Imani Fouladi AA, Farokh P. Study of aac(6')ie-aph(2'')ia gene in clinical strain of enterococci and identification of high-level gentamicin resistante enterococci. Scientific Journal of Hamadan University of Medical Sciences and Health Services 2010; 17: 25-32 (In Persian).
Feizabadi MM, Shokrzadeh L, Sayady S, Asadi S.Transposon Tn5281 is the main distributor of the aminoglycoside modifying enzyme gene among isolates of Enterococcus faecalis in Tehran hospitals. Can J Microbiol 2008; 54: 887-890.
Vigani AG, Macedo de Oliveira A, Bratfich OJ, Stucchi RSB, Moretti ML. Clinical, epidemiological, and micro- biological characteristics of bacteremia caused by high- level gentamicin-resistant Enterococcus faecalis. Braz J Med Biol Res 2008; 41: 890-895.
Vakulenko SB, Donabedian SM, Voskresenskiy AM, Voskresenskiy AM, Zervos MJ, Lerner SA, et al. Multiplex PCR for detection of aminoglycoside resistance genes in enterococci.Antimicrob Agents Chemother 2003; 47: 1423-1426.
Sahm DF, Gilmore MS. Transferability and genetic relatedness of high-level gentamicin resistance among enterococci. Antimicrob Agents Chemother 1994; 38:1194-1196.
Keddy KH, Klugman KP, Liebowitz LD. Incidence of high-level gentamicin resistance in enterococci at Johannesburg Hospital. S Afr Med J 1996; 86 :1273-1276.
Papaparaskevas J, Vatopoulos A, Tassios PT, Avlami A, Legakis NJ, Kalapothaki V. Diversity among high- level aminoglycoside-resistant enterococci. J Antimicrob Chemother 2000; 45: 277-283.
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Issue | Vol 5 No 3 (2013) | |
Section | Articles | |
Keywords | ||
Dot-Blot hybridization Enterococci High-level gentamicin resistance (HLGR) Long PCR |
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