Molecular Characterization of Ciprofloxacin Resistant Escherichia coli from Ghana

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Israel Mensah- Attipoe
Japheth A. Opintan
Mercy J. Newman
Prince Pappoe- Ashong


Aim: This study aimed to characterize ciprofloxacin-resistance genes in clinical Escherichia coli isolates obtained from a six-month antimicrobial resistance (AMR) surveillance from Ghana.

Methods: Eighty-three of 440 archived E. coli isolates were confirmed by biochemical reactions and resistance profiles by the disc diffusion method. These isolates were cultured from urine (42), stool (23), vaginal swabs (12), wounds (5) and heart valve (1) during AMR surveillance. Minimum Inhibition Concentration (MIC) by E-test method was performed on all E. coli isolates that were resistant to ciprofloxacin by the disc diffusion method. Additionally, all isolates with reduced MIC to ciprofloxacin (>32 µg/ml) were selected for molecular assays.  Three chromosomal and nine plasmid-mediated resistance genes were screened in all Ciprofloxacin resistant E. coli (CRE) by polymerase chain reaction (PCR). Randomly selected amplified genes were commercially sequenced and analyzed.

Results: In total, 47/83 (56.6%) E. coli isolates were resistant to ciprofloxacin and 29 (61.7%) had MIC values greater than 32 µg/ml. Chromosomal mediated genes (gyrA, gyrB and parC) were present in all 29 CRE isolates (100%). Distribution of the plasmid-mediated genes were as follows; qnrA 16/29 (55.1%), qnrB 16/29(55.1%), qnrC 22/29(75.8%), qnrS 26/29(89.6%), qepA 5/29(17.2%) and oqxB 19/29(65.5%). Genes encoding for altered aminoglycoside acetyltransferase [aac(6’)1bcr] were also present in all 29 CRE isolates. The majority (72.4%) of the CRE isolates had gyrA mutations at codons 83 and 87. In parC, the mutations were at codons 71 and 80. Five isolates had mutations at codon 56 and four each had mutations at positions 79 and 80.

Conclusion: In this study, fluoroquinolone resistance genes were identified in all CRE isolates, mostly with putative mutations in the Quinolone Resistance Determining Region (QRDR). These chromosomal and plasmid-mediated genes may be widespread in Ghana and associated with CRE from the AMR surveillance. Although new mutations points were identified in parC, they may not be linked to the CRE.

Antimicrobial resistance, antimicrobial sensitivity testing, resistance, ciprofloxacin resistant.

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How to Cite
Attipoe, I. M.-, Opintan, J. A., Newman, M. J., & Ashong, P. P.-. (2020). Molecular Characterization of Ciprofloxacin Resistant Escherichia coli from Ghana. Journal of Advances in Microbiology, 20(10), 22-33.
Original Research Article


Aarestrup FM, Wegener HC, Collignon P. Resistance in bacteria of the food chain: epidemiology and control strategies. Expert Review of Anti-infective Therapy. 2008;6(5):733-750.

Elliott E, Brink AJ, van Greune J, Els Z, Woodford N, Turton J, Livermore DM. In vivo development of ertapenem resistance in a patient with pneumonia caused by Klebsiella pneumoniae with an extended-spectrum β-lactamase. Clinical infectious Diseases. 2006;42(11): 95-98.

Giguère S. Antimicrobial drug action and interaction: An introduction. Antimicrobial Therapy in Veterinary Medicine. 2013;1-10.

Palmer KL, Kos VN, Gilmore MS. Horizontal gene transfer and the genomics of enterococcal antibiotic resistance. Current Opinion in Microbiology. 2010; 13(5):632-639.

Bellido F, Pechère JC. Laboratory survey of fluoroquinolone activity. Reviews of Infectious Diseases. 1989;11(Supplement _5):917-924.

Schaeffer AJ. The expanding role of fluoroquinolones. Disease-a-Month. 2003; 49(2):129-147.

Walsh F, Rogers TR. Comparison of plasmid-mediated quinolone resistance and extended-spectrum b-lactamases in third-generation cephalosporin-resistant Enterobacteriaceae from four Irish hospitals. Journal of Medical Microbiology. 2012;61:142-147.

Pitout JD, Multiresistant enterobacteria-ceae: New threat of an old problem. Expert Review of Anti-infective Therapy. 2008; 6(5): 657-669.

Puig C, Tirado-Vélez JM, Calatayud L, Tubau F, Garmendia J, Ardanuy C, Liñares J. Molecular characterization of fluoroquinolone resistance in nontypeable Haemophilus influenzae clinical isolates. Antimicrobial Agents and Chemotherapy. 2015;59(1):461-466.

Martínez-Martínez L, Pascual A, Jacoby GA. Quinolone resistance from a transferable plasmid. The Lancet. 1998; 351(9105):797-799.

Strahilevitz J, Jacoby GA, Hooper DC, Robicsek A. Plasmid-mediated quinolone resistance: a multifaceted threat. Clinical Microbiology Reviews. 2009;22(4):664-689.

Opintan JA, Newman MJ, Arhin RE, Donkor ES, Gyansa-Lutterodt M, Mills-Pappoe W. Laboratory-based nationwide surveillance of antimicrobial resistance in Ghana. Infection and Drug Resistance. 2015;8:379.

Rezazadeh M, Baghchesaraei H, Peymani A. Plasmid-mediated quinolone-resistance (qnr) Genes in clinical isolates of Escherichia coli collected from several hospitals of Qazvin and Zanjan Provinces, Iran. Osong Public Health and Research Perspectives. 2016;7(5):307-312.

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(10):2380-2386.

Wirth T, Falush D, Lan R, Colles F, Mensa P, Wieler LH. Ochman H. Sex and virulence in Escherichia coli: An evolutionary perspective. Molecular Microbiology. 2006; 60(5):1136- 1151.

Lane D. 16S/23S rRNA sequencing. Nucleic acid techniques in bacterial systematics. 1991;115-175.

Wang M, Guo Q, Xu X, Wang X, Ye X, Wu S., Wang M. New plasmid-mediated quinolone resistance gene, qnrC, found in a clinical isolate of Proteus mirabilis. Antimicrobial Agents and Chemotherapy, 2009;53(5):1892-1897.

Jacoby G, Cattoir V, Hooper D, Martínez-Martínez L, Nordmann P, Pascual A, Wang M. qnr gene nomenclature. Antimicrobial Agents and Chemotherapy. 2008;52(7): 2297-2299.

Cavaco LM, Hasman H, Xia S, Aarestrup FM. qnrD, a novel gene conferring transferable quinolone resistance in Salmonella enterica serovar Kentucky and Bovismorbificans strains of human origin. Antimicrobial Agents and Chemotherapy. 2009;53(2):603-608.

Périchon B, Courvalin P, Galimand M, Transferable resistance to aminoglycosides by methylation of G1405 in 16S rRNA and to hydrophilic fluoroquinolones by QepA-mediated efflux in Escherichia coli. Antimicrobial Agents and Chemotherapy. 2007;51(7):2464-2469.

Şahinturk P, Arslan E, Büyükcangaz E, Sonal S, Şen A, Ersoy F,. Cengiz M. High level fluoroquinolone resistance in Escherichia coli isolatedfrom animals in Turkey is due to multiple mechanisms. Turkish Journal of Veterinary and Animal Sciences. 2016;40(2):214-218.

Park CH, Robicsek A, Jacoby GA, Sahm D, Hooper DC. Prevalence in the United States of aac (6′)-Ib-cr encoding a ciprofloxacin-modifying enzyme. Antimicrobial Agents and Chemotherapy. 2006;50(11):3953-3955.

Al-Agamy M, Zaki SA. Mechanisms of fluoroqinolones resistance in Escherichia coli isolates from Saudi Arabia. African J Microb Res. 2012;6(1):155-9.

Jadoon RJ, Jalal-Ud-Din M, Khan SA. E. coli resistance to ciprofloxacin and common associated factors. Journal of the College of Physicians and Surgeons--Pakistan: JCPSP. 2015;25(11):824-827.

Foster S. The economic burden of antimicrobial resistance in the developing world, in antimicrobial resistance in developing countries. Springer. 2010;365-384.

Livermore DM, Hope R, Reynolds R, Blackburn R, Johnson AP, Woodford N, Declining cephalosporin and fluoroquinolone non-susceptibility among bloodstream Enterobacteriaceae from the UK: links to prescribing change? Journal of Antimicrobial Chemotherapy. 2013;68(11): 2667-2674.

Zhao G, Zhan X. Facile preparation of disposable immunosensor for Shigella flexneri based on multi-wall carbon nanotubes/chitosan composite. Electrochimica Acta. 2010;55(7):2466-2471.

Robicsek A, Jacoby GA, Hooper DC. The worldwide emergence of plasmid-mediated quinolone resistance. The Lancet Infectious Diseases. 2006;6(10): 629-640.

Tarchouna M, Ferjani A, Marzouk M, Guedda I, Boukadida J. Prevalence of plasmid-mediated quinolone resistance detrminants among clinical isolates of Escherichia coli in a Tunisian hospitals. Int. J. Curr. Microbiol. App. Sci. 2015;4(3): 195-206.

Machuca J, Ortiz M, Recacha E, Díaz-De-Alba P, Docobo-Perez F, Rodríguez-Martínez JM, Pascual Á. Impact of AAC (6′)-Ib-cr in combination with chromosomal-mediated mechanisms on clinical quinolone resistance in Escherichia coli. Journal of Antimicrobial Chemotherapy. 2016;71(11):3066-3071.

Jaktaji RP, Mohiti E. Study of mutations in the DNA gyrase gyrA gene of Escherichia coli. Iranian Journal of Pharmaceutical Research: IJPR. 2010;9(1):43.

Yoshida H, Bogaki M, Nakamura M, Yamanaka LM, Nakamura S. Quinolone resistance-determining region in the DNA gyrase gyrB gene of Escherichia coli. Antimicrobial Agents and Chemotherapy. 1991;35(8):1647-1650.

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(4):879-885.

Shoji H, Shirakura T, Fukuchi K, Takuma T, Hanaki H, Tanaka K, Niki Y. A molecular analysis of quinolone-resistant Haemophilus influenzae: Validation of the mutations in quinolone resistance-determining regions. Journal of Infection and Chemotherapy. 2014;20(4):250-255.

Soussy CJ, Wolfson JS, Ng EY, Hooper DC. Limitations of plasmid complementation test for determination of quinolone resistance due to changes in the gyrase A protein and identification of conditional quinolone resistance locus. Antimicrobial Agents and Chemotherapy. 1993;37(12):2588-2592.