Molecular characterization of class 1 integrons and gene cassettes in multidrug resistant (MDR) Klebsiella spp. isolated from hospitalized and outpatients in Iran, 2009
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Abstract
Background and objectives: Klebsiella species are of the most common bacteria involved in nosocomial and urinary tract infections. Genetic elements such as class 1 integrons have an important role in the resistance development. In this study, the share of class 1 integrons, the genetic characterization of the integron cassettes and PFGE profiles of the clinical Klebsiella isolates are evaluated in Besat University hospital of Sanandaj, Iran.
Methods: Isolates from 17890 clinical specimens were identified by API20E. Antibiotic susceptibility testing and MIC were done for MDR isolates. For investigating class 1 integrons and gene cassettes, PCR by intI1 integrase and 5΄-CS/3΄-CS were performed. Integrated gene cassettes were analyzed by PCR-RFLP and sequencing. Pulsed-Field Gel Electrophoresis was carried out for studying of clonality outbreak of isolates.
Results: Thirty five Klebsiella spp. were isolated and included 29 K. pneumoniae and six K. oxytoca. All the isolates were susceptible to carbapenems while other antibiotics showed high resistant profile. In all Klebsiella spp. PCR for intI1 integrase and 5΄-CS/3΄-CS were positive (100%). Sequencing for prevalent bands of internal variable regions between 5΄-CS/3΄-CS showed arr-5, orfD-aacA4 and aad5- dfrA17. PFGE Analysis showed 18 clusters in K. pneumoniae with clonality relatedness in some cases but no relatedness among K. oxytoca isolates.
Conclusion: High prevalence of class 1 integron carrying gene cassettes confirms that integron-mediated antimicrobial gene cassettes are important in Klebsiella spp. resistance profile. Clone diffusions of MDR Klebsiella spp. which harbor class 1 integrons have threaten the potential in the resistance development in our clinical settings.
Damjanova I,Tóth A, Pászti J, Hajbel-Vékony G, Jakab M, Berta J, et al. Expansion and countrywide dissemination of ST11, ST15 and ST147 ciprofloxacin- resistant CTX-M-15-type b-lactamase-producing Klebsiella pneumoniae epidemic clones in Hungary in 2005—the new ‘MRSAs’? J Antimicrob Chemther 2008; 62: 978-985.
Yu VL, Hansen DS, Ko WC, Sagnimeni A, Klugman KP, von Gottberg A, et al. International Klebseilla Study Group. Virulence Characteristics of Klebsiella and Clinical Manifestations of K. pneumoniae Bloodstream Infections. Emerg Infect Dis 2007; 13: 986-993.
Blot S, Depuydt P, Vandewoude K, DeBacquer D.
Measuring the impact of multidrug resistance in nosocomial infection. Curr Opin Infect Dis 2007; 20:391-396, Review.
Biedenbach DJ, Moet GJ, Jones RN. Jones. Occurrence and antimicrobial resistance pattern comparisons among bloodstream infection isolates from SENTRY Antimicrobial Surveillance Program. Diagn Microbiol Infect Dis 2004; 50: 59-69.
Roy Chowdhury P, Ingold A, Vanegas N, Martínez E, Merlino J, Merkier AK, et al. Dissemination of multiple drug resistance genes by class 1 integrons in Klebsiella pneumoniae isolates from four countries: a comparative study. Antimicrob Agents Chemother 2011; 55: 3140-3149.
Betteridge T, Partridge SR, Iredell JR, Stokes HW.
Genetic context and structural diversity of class 1 integrons from human commensal bacteria in a hospital intensive care unit. Antimicrob Agents Chemother 2011;55: 3939-3943.
Hall RM, Collis CM. Antibiotic resistance in Gram- negative bacteria: the role of gene cassettes and integrons. Drug Resist Updat 1998; 1: 109-119.
Rowe-Magnus DA, Mazel D. Resistance gene capture.Curr Opin Microbiol 1999; 2: 483-488.
Cambray G, Guerout AM, Mazel D. Integrons. Annu Rev Genet 2010; 44: 141-166.
Partridge SR, Tsafnat G, Coiera E, Iredell JR. Gene cassettes and cassette arrays in mobile resistance integrons. FEMS Microbiol Rev 2009; 33: 757-784.
Pop-Vicas A, Strom J, Stanley K, D’Agata EM. Multidrug-resistant Gram-negative bacteria among patients who require chronic hemodialysis. Clin J Am Soc Nephrol 2008; 3: 752-758.
Farmer JJ (2003). Enterobacteriaceae: introduction and identification. In: Murray PR, Baron EJ, Jorgensen JH, Pfaller MA, Yolken RH, editors. Manual of clinical microbiology. 8th ed. Washington: American Society for Microbiology, p. 636-653.
Clinical and Laboratory Standards Institute (CLSI) (2010). Performance Standards for Antimicrobial Susceptibility Testing, Twenty-First Informational Supplement M100-S20, Vol. 31, No. 1, Wayne.
Andrews JM. Determination of Minimum Inhibitory Concentration, J Antimicrob Chemther 2001; 48 : 5-16.
Rosser SJ. Young HK. Identification and characterization of class 1 integrons in bacteria from an aquatic environ ment. J Antimicrob Chemother 1999; 44: 11-18.
Levesque C, Piche L, Larose C, Roy PH. PCR mapping of integrons reveals several novel combinations of resistance genes. Antimicrob Agents Chemother 1995;39: 185-191.
Shahcheraghi F, Badmasti F, Feizabadi MM.Molecular characterization of class 1 integrons in MDR Pseudomonas aeruginosa isolated from clinical settings in Iran, Tehran. FEMS Immunol Med Microbiol 2010;58: 421-425.
Centers for Disease Control and Prevention. Standardized Molecular Subtyping of Foodborne Bacterial Pathogens by Pulsed-Field Gel Electrophoresis: Training Manual. Atlanta, GA: 2000.
Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, Swaminathan B. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol 1995; 33: 2233-2239.
Hansen DS, Gottschau A, Kolmos HJ. Epidemiology of Klebsiella bacteraemia: a case control study using Escherichia coli bacteraemia as control. J Hosp Infect 1998; 38: 119-132.
Akinci E, Colpan A, Bodur H, Balaban N, Erbay A.Risk factors for ICU-acquired imipenem-resistant Gram-negative bacterial infections. J Hosp Infect 2005;59: 317-323.
Ramazanzadeh R. Prevalence and characterization of extended-spectrum beta-lactamase production in clinical isolates of Klebsiella spp. Afr J Microbiol Res 2010; 4;1359-1362.23. 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: 247-250.
Feizabadi MM, Delfani S, Raji N, Majnooni A, Aligholi M, Shahcheraghi F, Parvin M, Yadegarinia D. Distribution of bla(TEM), bla(SHV), bla(CTX-M) genes among clinical isolates of Klebsiella pneumoniae at Labbafinejad Hospital, Tehran, Iran. Microb Drug Resist 2010; 16: 49-53.
Feizabadi MM, Mahamadi-Yeganeh S, Mirsalehian A, Mirafshar SM, Mahboobi M, Nili F, et al. Genetic characterization of ESBL-producing Klebsiella pneumoniae J Infect Dev Ctries 2010; 4: 609-615.
Ghafourian et al. The Prevalence of ESBLs Producing Klebsiella pneumoniae Isolates in some Major Hospitals, Iran. Open Microb J 2011; 5.
Nematzadeh S, Shahcheraghi F, Feizabadi MM, Nikbin VS, Nasehi L. Molecular characterization of CTX- Mβ-lactamases among Klebsiella pneumoniae isolated from patients at Tehran hospitals. Indian J Med Microbiol 2011; 29: 254-257.
Shahcheraghi F, Nobari S, Rahmati Ghezelgeh F, Nasiri S, Owlia P, Nikbin VS, Imani Fooladi AA. First Report of New Delhi Metallo-Beta-Lactamase-1-Producing Klebsiella pneumoniae in Iran. Microb Drug Resist 2012 17. [Epub ahead of print]
Yousefi S, Nahaei MR, Farajnia S, Ghojazadeh M, Akhi MT, Sharifi Y, Milani M, Ghotaslou R. Class 1 integron and Imipenem Resistance in Clinical Isolates of Pseudomonas aeruginosa: Prevalence and Antibiotic Susceptibility. Iran J Microbiol 2010; 2: 113-119.
Firoozeh F, Shahcheraghi F, Zahraei Salehi T, Karimi V, Aslani MM. Antimicrobial resistance profile and presence of class I integrongs among Salmonella enterica serovars isolated from human clinical specimens in Tehran, Iran. Iran J Microbiol 2011; 3: 112-117.
Firoozeh F, Zahraei-Salehi T, Shahcheraghi F, Karimi V, Aslani MM. Characterization of class I integrons among Salmonella enterica serovar Enteritidis isolated from humans and poultry. FEMS Immunol Med Microbiol 2012; 64: 237-243.
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| Issue | Vol 5 No 1 (2013) | |
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| Keywords | ||
| Klebsiella spp. class 1 integrons multidrug resistante (MDR) | ||
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