The Survey of Genes Encoding Beta-Lactamases, in Escherichia Coli Resistant to Beta-Lactam and Non-Beta-Lactam Antibiotics

Document Type : Original Article

Authors

Department of Microbiology, Pasteur Institute of Iran, Tehran, Ira

Abstract

Objective(s)
Resistance to the new generation of cephalosporins which is mediated by Extended-Spectrum beta-lactamases (ESBLs) has been found among Escherichia coli isolates throughout the world. These resistance genes and their producers, the micro-organisms carrying beta-lactamases, are responsible for serious clinical and therapeutic problems among inpatients and it is necessary to pay more attention to detection of ESBLs producing organisms.
Materiasl and Methods
Collectively 260 isolates of E. coli were obtained from 6 hospitals in Tehran (Iran) during April-2006 to April-2007. The antibiotic susceptibility patterns of isolates were determined by disk diffusion method. phenotypic confirmatory test (PCT) was carried out for screening of ESBLs. Microbroth dilution assay was used to determine the minimum inhibitory concentration (MIC) of ceftazidime. Isolates showing MIC>2 pg/ml were subjected to polymerase chain reaction (PCR) targeting blaTEM, blaSHV, blaCTX and blaPER genes.
Results
The PCT showed that 48.08% of isolates are ESBL producers (125 of 260). The majority of cefotaxime resistant (90.8%) and ceftazidime resistant (92.5%) isolates were ESBL producers. The obtained results by PCR revealed that 5.77% (n=15 of 260) and 24.23 (n=63) of isolates can produce SHV and TEM type enzymes respectively. blaCTX was detected in 20.38% of isolates (n=53) and none of them could produce blaPER type beta-lactamases.
Conclusion
The results of our study showed that the ESBL genes have high prevalence among clinical isolates of E. coli. Such high dissemination of ESBLs is a serious problem for public health and therefore, it's necessary to seek a program for monitoring ESBLs in hospitals.

Keywords


1.Branger C, Zamfir O, Geoffroy S, Laurans G, Arlet G, Thien HV, et al. Genetic bachground of Escherichia coli and extended-spectrum beta-lactamase type, Imerging Infectious Disease. 2005.
2.Forward KR. Extended-spectrum beta-lactamases. 2006; 17:6B-8B.
3.Jeong SH, Bae IK, Lee JH, Sohn SG, Kang GH, Jeon GJ, et al. Molecular characterization of extended-spectrum beta- lactamases produced by clinical isolates of Klebsiella pneumoniae and Escherichia coli from a Korean nationwide survey. J Clin Microbiol 2004; 42:2902-2906.
4.Sanders CC, Sanders WE JR. Beta-lactamase in gram-negative bacteria: global trends and clinical impact. Clin Infect Dis 1992; 15:824-839.
5.Du Bois SK, Marriott MS, Amyes SGB. TEM-and SHV-derived extended spectrum B-lactamases: relationship between selection, structure and function. J Antimicrob Chemother 1995; 35:7-22.
6.Winokur PL, Canton R, Casellas JM, Legakis N.Variations in the prevalence of strains expressing an extended- spectrum beta-lactamase phenotype and characterization of isolates from Europe, the Americas, and the Western Pacific region. Clin infect dis 2001; 32:S94-103.
7.Cartelle M, del Mar Tomas M, Molina F, Moure R, Villanueva R, Bou G. High level resistance to ceftazidime conferred by a novel enzyme, CTX-M-32, derived from CTX-M-1 through a single Asp 240-Gly substitution. Antimicrob Agents Chemother 2004; 48:2308-2313.
8.Tzouvelekis LS, Tzelepi E, Tassios PT, Legakis NJ. CTX-M-type betalactamases: an emerging group of extended-spectrum enzymes. Int J Antimicrob Agents 2000; 14:137-142.
9.Weldhagen GF, Poirel L, Nordman P. Ambler class a extended-spectrum beta-lactamases in Pseudomonas aeroginosa-novel developments and clinical impact. Antimicrob Agents Chemother 2003; 47:2385-2392.
10.Shahcheraghi F, Moezi H, Feizabadi MM. Distribution of TEM and SHV Beta-lactamase genes among Klebsiella pneumoniae strains isolated from patients in Tehran. Med Sci Monit 2007; 13:BR247-250. 17968291.
11.Podschun R, Ullmann U. Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev 1998; 11:589-603.
12.National Committee for Clinical Laboratory Standards, Performance standards for antimicrobial susceptibility testing, 12th informational supplement. M100-S12. National Committee for Clinical Laboratory Standards.Wayne:Pa.2002.
13.Schwaber MJ, Raney PM, Raney PM, Rasheed JK, Biddle JW, Williams P, et al. Utility of NCCLS guidelines for identifying extended-spectrum beta-lactamases in non-Escherichia coli and Non-Klebsiella spp. of Enterobacteriaceae. J Clin Microbiol 2004; 42:294-298.
14.Detection of Extended-Spectrum beta-Lactamases (ESBL) in Escherichia coli and Kelebsiella species, British Society for Antimicrobial Chemotherapy.
15.National Committee for Clinical Laboratory Standards, Performance standards for antimicrobial susceptibility testing, 15th informational supplement (M100-S15), National Committee for Clinical Laboratory Standards. Wayne: Pa.2005.
16.Hammond DS, Schooneveldt JM, Nimmo GR, Huygens F, Giffard PM. blaSHV genes in Klebsiella pneumoniae different allele distributions are associated with different promoters within individual isolates .Antimicrob Agents Chemother 200; 49:256-263.
17.Weldhagen GF, Poirel L, Nordmann P. Ambler lass a extended-spectrum beta-lactamases in Pseudomonas aeruginosa: novel developments and clinical impact. Antimicrob Agents Chemother 2003; 47:2385-2392.
18.Bonnet R, Dutour C, Sampaio JL, Chanal C, Sirot D, Labia R, et al. Novel cefotaximase (CTX-M-16) with increased catalytic efficiency due to substitution Asp-240Gly. Antimicrob Agents Chemother 2001; 45:2269-22675.
19.Danel F, Hall LM, Gur D, Akalin HE, Livermore DM. Transferable production of PER-1 beta-lactamase in Pseudomonas aeruginosa.J. Antimicrob Chemother 1995; 35:281-294.
20.Howard C, Van Daal A, Van Daal A, Kelly G, Schooneveldt J, Nimmo G, et al. Identification and minisequencing-based discrimination of SHV beta-lactamases in nosocomial infection-associated Klebsiella pneumoniae in Brisbane, Australia. Antimicrob Agents Chemother 2002; 46:659-664.
21.Chambers PA, Duggan PS, Heritage J, Forbes JM. The fate of antibiotic resistance marker genes in transgenic plant feed material fed to chickens. J Antimicrob Chemother 2002; 49:161-164.
22.Edelstein M, Pimkin M, Palagin I, Edelstein I, Stratchounski L. Prevalence and Molecular Epidemiology of CTX-M Extended-Spectrum B-Lactamase-Producing Escherichia coli and Klebsiella pneumoniae in Russian Hospitals. Antimicrob Agents Chemother 2003; 47:3724-3732.
23.Bradford PA. Extended-spectrum beta-lactamases in the 21 st century: characterization, epidemiology, and detection of this important resistancethreat. Clin Microbiol Rev 2001; 14:933-951. 
24.Ben-Hamouda T, Foulon T, Ben-Cheikh-Masmoudi A, Fendri C, Belhadj O, Ben-Mahrez K. Molecular epidemiology of an outbreak of multiresistant Klebsiella pneomoniae in a Tunisian neonatal ward. J Med Microbiol 2003; 52:427-433.
25.Moubareck C, Daoud Z, Hakime NI, Hamze M, Mangeney N, Matta H, et al. Countrywide spread of community and hospital acquired extended-spectrum beta-lactamase (CTX-M-15) producing Enterobacteriaceae in Lebanon. J Clin Microbiol 2005; 43:3309-3313.
26.Kader AA, Kumar A. Prevalence and antimicrobial susceptibility of extendedspectrum beta-lactamase- producing Escherichia coli and Klebsiella pneumoniae in a general hospital. Ann Saudi Med 2005; 25:239-242.
27.Kjerulf A, Hansen DS, Sandvang D, Hansen F, Frimodt-Moller N. The prevalence of ESBL-producing E. coli and Klebsiella strains in the Copenhagen area of Denmark. APMIS 2008; 116:118-124.
28.Ko KS, Lee MY, Song JH, Lee H, Jung DS, Jung SI, et al. Prevalence and characterization of extended-spectrum beta-lactamase-producing Enterobacteriaceae isolated in Korean hospitals. Diagn Microbiol Infect Dis 2008; 13.
29.Ta§li H, Bahar IH. Mulecular characterization of TEM and SHV derived extended-spectrum beta-lactamases in hospital based enterobacteriaceae in Turkey. Jpn J Infect Dis 2005; 58:162-167.
30.Bean DC, Krahe D, Wareham DW. Antimicrobial resistance in community and nosocomial Escherichia coli urinary tract isolates, London 2005 - 2006. Ann Clin Microbiol Antimicrob 2008; 7:13.
31.Ananthan S, Subha A. Cefoxitin resistance mediated by loss of a porin in clinical strains of Klebsiella pneumoniae and Escherichia coli. Indian J Med Microbiol 2005; 23:20-23.
32.Seema singh, SCM (ASM). Eetended-Spectrum beta-lactamases (ESBLs): An Overview. 1999.
33.Bus K, Jacoby GA, Medeiros AA. A functional classification scheme for beta-lactamases and its correlation with molecular structure. Antimicrob Agents Chemother 1995; 39:1211-1233.
34.Rodriguez-Bano J, Navarro MD, Romero L, Martinez-Martinez L, Muniain MA, Perea EJ, et al. Epidemiology and clinical features of infections caused by extended-spectrum beta-lactamase-producing Escherichia coli in nonhospitalized patients. J Clin Microbiol 2004; 42:1089-1094.
35.Conceicao T, Brizio A, Duarte A, Barros R. First isolation of bla (VIM-2) in Klebsiella oxytoca clinical isolates from Portugal, Antimicrob. Agents Chemother 2005; 49:476.
36.Giakkoupi P, Tzouvelekis LS, Daikos GL, Miriagou V, Petrikkos G, Legakis NJ, et al. Discrepancies and interpretation problems in susceptibility testing of VIM-1-producing Klebsiella pneumoniae isolates. J Clin Microbiol 2005; 43:494-496.
37.Koh TH, Sng LH, Babini GS, Woodford N, Livermore DM, Hall LM. Carbapenem-resistant Klebsiella pnuemoniae in singapore producing IMP-1 betalactamase and lacking an outer membrane protein,Antimicrob. Agents Chemother 2001; 45:1939-1940.
38.Matar GM, Al Khodor S, El-Zaatari M, Uwaydah M. Prevalence of the genes encoding extended-spectrum beta-lactamases, in Escherichia coli resistant to beta-lactamand non-beta-lactam antibiotics. Ann Trop Med Parasitol 2005; 99:413-417.
39.Ariffin H, Navaratnam P, Mohamed M, Arasu A, Abdullah WA, Lee CL, et al. Ceftazidime-resistant Klebsiella pneumoniae bloodstream infection in children with febrile neutropenia. Int J Infect Dis 2000; 4:21-25.
40.Jacoby GA, Carreras I. Activities of P-Lactam antibiotics against Escherichia coli strains producing extended- spectrum beta-lactamases, antimicrob. Agents Chemother 1990; 34:858-862.
41.Chmelnitsky I, Carmeli Y, Leavitt A, Schwaber MJ, Navon-Venezia S. CTX-M-2 and a New CTX-M-39 enzyme are the major extended-spectrum beta-lactamases in multiple Escherichia coli clones isolated in Tel Aviv, Israel. Antimicrob Agents Chemother 2005; 49:4745-4750.
42.Fazly Bazzaz BS, Naderinasab M, Mohamadpoor AH, Farshadzadeh Z, Ahmadi S, Yousefi F. The prevalence of extended-spectrum beta lactamase-producing Escherichia coli and Klebsiella pneumoniae among clinical isolates from a general hospital in Iran. Acta Microbiologica et Immunologica Hungarica, 2009; 56:89-99. DOI: 10.1556/AMicr.56.2009.1.7
43.Hernandez JR, Martinez-Martinez L, Canton R, Coque TM, Pascual A. Nationwide study of Escherichia coli and Klebsiella pneumoniae producing extended-spectrum beta-lactamases in Spain. Antimicrob Agents Chemother 2005; 49:2122-2125.
44.Kiratisin P, Apisarnthanarak A, Laesripa C, Saifon P. Molecular characterization and epidemiology of extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae causing health care- associated infection in Thailand: An endemic area of CTX-M. Antimicrob Agents Chemother 2008; 52:2818-24.
45.Katz OT, Peled N, Yagupsky P. Evaluation of the current national committee for clinical laboratory standards guidelines for screeningand confirming extended-spectrum beta-lactamase production in isolates of Escherichia coli and Klebsiella species from bacteremic patients. Eur J Clin Microbiol Infect Dis 2004; 23:813-817.
46.Lautenbach E, Patel JB, Bilker WB, Edelstein PH, Fishman NO. Extendedspectrum beta-lactamase-producing Escherichia coli and Klebsiella-pneumoniae:risk factors for infection and impact of resistance on outcomes. Clin Infect Dis 2001; 32:1162- 1171.
47.Livermore DM. Determinants of the activity of beta-lactamase inhibitor combinations. J Antimicrob Chemother 1993; 31:9, 21.
48.Lucet JC, Regnier B. Enterobacteria producing extended-spectrum betalactamases. Pathol Biol 1998; 46:235-243.