Background: Fluoroquinolone resistant Escherichia coli isolates have become an important challenge in healthcare settings in Iran. In this study, we have determined Fluoroquinolone resistant E.coli isolates (from both outpatients and inpatients) and evaluated mutations of gyrA and parC within the quinolone resistance-determining regions (QRDR) of these clinical isolates. Materials and Method: A total of 135 E.coli clinical isolates were recovered from urine of 135 patients (91 outpatients and 44 inpatients) admitted at Alzahra hospital, Iran, between September and February 2013. We assessed antimicrobial susceptibility of all isolates and determined mutations in QRDR of gyrA and parC genes from 13 fluoroquinolone-resistant isolates by DNA sequencing. Results: In this study resistance rate of fluoroquinolones (Ciprofloxacin, Norfloxacin and Ofloxacin) were 45.2%. Two E.coli isolates were shown just a single mutation, but other isolates possessed 2,3,4 and 5 mutations in gyrA and parC genes. Mutations in the QRDR regions of gyrA were at positions Ser83 and Asp87 and parC at positions Ser80, Glu84, Gly78. Conclusions: Ciprofloxacin is the most common antimicrobial agent used for treating urinary tract infections (UTIs) in healthcare settings in Iran. Accumulation of different substitutions in the QRDR regions of gyrA and parC confers high-level resistance of fluoroquinolones in clinical isolates.
Momtaz H, Karimian A, Madani M, Safarpoor Dehkordi F, Ranjbar R, Sarshar M, et al. Uropathogenic Escherichia coli in Iran: serogroup distributions, virulence factors and antimicrobial resistance properties. Ann Clin Microbiol Antimicrob. 2013;12.
Kucheria R, Dasgupta P, Sacks S, Khan M, Sheerin N. Urinary tract infections: new insights into a common problem. Postgraduate medical journal. 2005;81:83.
Wiles TJ, Kulesus RR, Mulvey MA. Origins and virulence mechanisms of uropathogenic< i> Escherichia coli. Experimental and molecular pathology. 2008;85:11-9.
Farajnia S, Alikhani MY, Ghotaslou R, Naghili B, Nakhlband A. Causative agents and antimicrobial susceptibilities of urinary tract infections in the northwest of Iran. International Journal of Infectious Diseases. 2009;13:140-4.
Koningstein M, van der Bij AK, de Kraker ME, Monen JC, Muilwijk J, de Greeff SC, et al. Recommendations for the Empirical Treatment of Complicated Urinary Tract Infections Using Surveillance Data on Antimicrobial Resistance in the Netherlands. PloS one. 2014;9:e86634.
Jacoby GA. Mechanisms of resistance to quinolones. Clinical Infectious Diseases. 2005;41:S120-S6.
Yamane T, Enokida H, Hayami H, Kawahara M, Nakagawa M. Genome‐wide transcriptome analysis of fluoroquinolone resistance in clinical isolates of Escherichia coli. International Journal of Urology. 2012;19:360-8.
King DE, Malone R, Lilley SH. New classification and update on the quinolone antibiotics. American family physician. 2000;61:2741-8.
Drago L, De Vecchi E, Mombelli B, Nicola L, Valli M, Gismondo M. Activity of levofloxacin and ciprofloxacin against urinary pathogens. Journal of Antimicrobial Chemotherapy. 2001;48:37-45.
Oliphant CM and Green GM. Quinolones: a comprehensive review. American family physician. 2002;65:455-64.
Soltani R, Ehsanpoor M, Khorvash F, Shokri D. Antimicrobial susceptibility pattern of extended-spectrum β-lactamase-producing bacteria causing nosocomial urinary tract infections in an Iranian referral teaching hospital. Journal of Research in Pharmacy Practice. 2014;3:6.
Faghri J, Salehi-Abargouei A, Sedighi M, Oryan G. Epidemiology of VIM-1-imipenem Resistant Pseudomonasaeruginosa in Iran: A Systematic Review and Meta-analysis. Journal of Research in Medical Sciences. 2014;19.
Baral P, Neupane S, Marasini BP, Ghimire KR, Lekhak B, Shrestha B. High prevalence of multidrug resistance in bacterial uropathogens from Kathmandu, Nepal. BMC research notes. 2012;5:38.
Morgan-Linnell SK, Boyd LB, Steffen D, Zechiedrich L. Mechanisms accounting for fluoroquinolone resistance in Escherichia coli clinical isolates. Antimicrobial agents and chemotherapy. 2009;53:235-41.
Morgan-Linnell SK and Zechiedrich L. Contributions of the combined effects of topoisomerase mutations toward fluoroquinolone resistance in Escherichia coli. Antimicrobial agents and chemotherapy. 2007;51:4205-8.
Bansal S and Tandon V. Contribution of mutations in DNA gyrase and topoisomerase IV genes to ciprofloxacin resistance in< i> Escherichia coli clinical isolates. International journal of antimicrobial agents. 2011;37:253-5.
Tille P. Bailey & Scott's diagnostic microbiology: Elsevier Health Sciences. 2013.
Cockerill FR, Clinical, Institute LS. Performance standards for antimicrobial disk susceptibility testing: approved standard: National Committee for Clinical Laboratory Standards, 2012.
Green MR and Sambrook J. Molecular cloning: a laboratory manual: Cold Spring Harbor Laboratory Press New York,. 2012.
Ahmed OB, Asghar AH, Elhassan MM. Comparison of three DNA extraction methods for polymerase chain reaction (PCR) analysis of bacterial genomic DNA. African Journal of Microbiology Research. 2014;8:598-602.
Everett MJ, Jin YF, Ricci V, Piddock L. Contributions of individual mechanisms to fluoroquinolone resistance in 36 Escherichia coli strains isolated from humans and animals. Antimicrobial Agents and Chemotherapy. 1996;40:2380-6.
Hooper DC. Bacterial topoisomerases, anti-topoisomerases, and anti-topoisomerase resistance. Clinical infectious diseases. 1998;27:S54-S63.
Heisig P. Genetic evidence for a role of parC mutations in development of high-level fluoroquinolone resistance in Escherichia coli. Antimicrobial Agents and Chemotherapy. 1996;40:879-85.
Ito CAS, Gales AC, Tognim MCB, Munerato P, Dalla Costa LM. Quinolone-resistant Escherichia coli. Brazilian Journal of Infectious Diseases. 2008;12:5-9.
Gales A, Gordon K, Wilke W, Pfaller M, Jones R. Occurrence of single-point< i> gyr A mutations among ciprofloxacin-susceptible< i> Escherichia coli isolates causing urinary tract infections in Latin America. Diagnostic microbiology and infectious disease. 2000;36:61-4.
Ruiz J. Mechanisms of resistance to quinolones: target alterations, decreased accumulation and DNA gyrase protection. Journal of Antimicrobial Chemotherapy. 2003;51:1109-17.
Hakanen A, Kotilainen P, Jalava J, Siitonen A, Huovinen P. Detection of decreased fluoroquinolone susceptibility in salmonellas and validation of nalidixic acid screening test. Journal of clinical microbiology. 1999;37:3572-7.
Ruiz J, Gómez J, Navia MM, Ribera A, Sierra JM, Marco F, et al. High prevalence of nalidixic acid resistant, ciprofloxacin susceptible phenotype among clinical isolates of< i> Escherichia coli and other< i> Enterobacteriaceae. Diagnostic microbiology and infectious disease. 2002;42:257-61.
Asadi S, Kargar M, Solhjoo K, Najafi A, Ghorbani-Dalini S. The association of virulence determinants of uropathogenic Escherichia coli with antibiotic resistance. Jundishapur Journal of Microbiology. 2014;7.
Ghadiri H, Vaez H, Khosravi S, Soleymani E. The antibiotic resistance profiles of bacterial strains isolated from patients with hospital-acquired bloodstream and urinary tract infections. Critical care research and practice. 2012;2012.