Molecular Characterization of Some Virulence Genes and Antibiotic Susceptibility Pattern among Uropathogenic Escherichia coli Isolated from Patient in Zakho City/Iraq
Uropathogenic Escherichia coli (UPEC) is one of the most causative agents which causing urinary tract infections (UTIs) in humans. This study involved the prevalence of the virulence genes among UPEC isolated from patients in various hospitals in Zakho city from July 2018 until January 2019 and their susceptibility to different commonly used antimicrobial agent against UPEC. The different culture media were used for the identification of Escherichia coli (E. coli). Out of 400 samples, 141 (35.25%) strains of UPEC were isolated from enrolled patients. The antibiotic susceptibility toward different antibiotics was varied among the isolates. Imipenem was the most potent antibiotic with a resistant rate of only 2.84%. While the isolates were resistant to most screened antibiotics, with the highest rate 96.45% to Amoxicillin/ clavulanic. The resistant rates decreased toward other antibiotics at rates varied from 93.62% for Amoxicillin to 43.97% for Norfloxacin. Fifty-seven isolates were selected for PCR analysis, according to the resistance of E.coli to various antibiotics. The selected samples were successfully amplified for E. coli identification by producing a single band of a target uidA gene. In this study, the virulence related genes were detected in only 35 (61.40%) isolates out of 57 isolates. The distribution of the virulence related genes that included; afa, sfa, hly, cnf and pai were 28.07%, 17.54%, 26.32%, 22.81% and 22.81%, respectively. The study highlight that multidrug resistance UPEC harbors multiple virulence genes circulating in this setting.
Alexander, S.K. and Strete, D. 2001. Microbiology: a photographic atlas for the laboratory, USA, Daryl Fox.
Assafi, M.S.A., Ibrahim, N.M.R., Hussein, N.R., Taha, A.A. and Balatay, A.A. 2015. Urinary Bacterial Profile and Antibiotic Susceptibility Pattern among Patients with Urinary Tract Infection in Duhok City, Kurdistan Region, Iraq, International Journal of Pure & Applied Sciences & Technology, 30(2), 54–63.
Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. 2003. Current Protocols in Molecular Biology, USA, John Wiley & Sons.
Bari, M.A., Arefin, M., Nessa, M., Mostofa, G. and Islam, T. 2017. Recent Antibiotic Sensitivity Pattern of Escherichia coli in Urinary Tract Infection, Journal of Teachers Association, 30(1), 61–65.
Behzadi, P., Najafi, A., Behzadi, E. and Ranjbar, R. 2016. Microarray long oligo probe designing for Escherichia coli : An in-silico DNA marker extraction, Central European Journal of Urology, 69(1), 105–111.
Bien, J., Sokolova, O. and Bozko, P. 2012. Role of Uropathogenic Escherichia coli Virulence Factors in Development of Urinary Tract Infection and Kidney Damage, International Journal of Nephrology, 2012, 1–15.
Le-Bouguenec, C.L.E., Archambaud, M. and Labigne, A. 1992. Rapid and Specific Detection of the pap , afa , and sfa Adhesin- Encoding Operons in Uropathogenic Escherichia coli Strains by Polymerase Chain Reaction, Journal of Clinical Microbiology, 30 (5), 1189–1193.
Chapman, T.A., Wu, X., Barchia, I., Bettelheim, K.A., Driesen, S., Trott, D., Wilson, M. and Chin, J.J. 2006. Comparison of Virulence Gene Profiles of Escherichia coli Strains Isolated from Healthy and Diarrheic Swine, Enviromental Microbiology ,72(7), 4782–4795.
Cheesbrough, M. 2006. District Laboratory Practice in Tropical Countries, United States of America New York, Cambridge University Press.
Coyle, M., B. 2005. Manual of Antimicrobial Susceptibility Testing, USA, Washington, DC : American Society for Microbiology.
Dash, M., Padhi, S., Mohanty, I., Panda, P. and Parida, B. 2013. Antimicrobial resistance in pathogens causing urinary tract infections in a rural community of Odisha, India, Journal of Family and Community Medicine, 20(1), 20–26.
Van De, S.-B.N., Grundmann, H., Verloo, D., Tiemersma, E., Monen, J., Goossens, H., Ferech, M., System, Surveillance, E.A.R. and Groups, E.S. of A.C.P. 2008. Antimicrobial drug use and resistance in Europe, Antimicrobial Drug Use and Resistance in Europe drug, 14(11), 1722–1730.
Dobrindt, U., Agerer, F., Michaelis, K., Janka, A., Buchrieser, C., Samuelson, M., Svanborg, C., Gottschalk, G., Karch, H. and Hacker, J. 2003. Analysis of Genome Plasticity in Pathogenic and Commensal Escherichia coli Isolates by Use of DNA Arrays, Journal of Bacteriology, 185(6), 1831–1840.
Dobrindt, U., Blum-oehler, G., Nagy, G., Schneider, G., Johnn, A. and Gottschalk, G. 2002. Genetic Structure and Distribution of Four Pathogenicity Islands (PAI I 536 to PAI IV 536 ) of Uropathogenic, American Society for Microbiology, 70(11), 6365–6372.
Engelkirk, P.G. and Duben-Engelkirk, J. 2008. Laboratory diagnosis of Infectious Diseases, USA, Wolters Kluwer Health/Lippincott Williams & Wilkins.
Farell, D.J., Morrissay, I., De Rubeids, D., Robbins, M. and Felmingham, D. 2003. A UK multicenter study and the antimicrobial susceptibility of bacterial pathogens causing urinry tract infection, Journal of Infection, 46 (2), 94–100.
Farshad, S., Ranjbar, R., Japoni, A., Hosseini, M., Anvarinejad, M. and Mohammadzadegan, R. 2012. Microbial Susceptibility, virulence Factors, and Plasmid Profiles of Uropathogenic Escherichia coli Strains Isolated from children in Jahrom, Iran. Arch Iran Med, 15(5), 312–316.
Firoozeh, F., Saffari, M., Neamati, F. and Zibaei, M. 2014. Detection of virulence genes in Escherichia coli isolated from patients with cystitis and pyelonephritis, International Journal of Infectious Diseases, 29, 219–222.
Gupta, K., Hooton, T.M., Naber, K.G., Wullt, B., Colgan, R., Miller, L.G., Moran, G.J., Nicolle, L.E., Raz, R., Schaeffer, A.J. and Soper, D.E. 2011. Highlights from international clinical practice guidelines for the treatment of acute uncomplicated cystitis and pyelonephritis in women: A 2010 update by the infectious diseases society of America and the european society for microbiology and infectious, Clin Infect Dis., 52(5), e103–e120.
Hassan, S.A., Jamal, S.A.,and Kamal, M. 2011. Occurrence of multidrug resistant and ESBL producing E. coli causing urinary tract infections, Journal of Basic & Applied Sciences, 7(1), 39–43.
Hegazy, E.E., Alam El Din, R.A.E., Amin, A.M., Mahgoub, F.M. and El Gamal, S.A.E. 2018. Microbiological Profile of Urinary Tract Infections with special Reference to Antibiotic Susceptibility Pattern of Escherichia coli Isolates, International Journal of Current Microbiology and Applied Sciences, 7(2), 911–920.
Hryniewicza, K., Szczypab, K., Sulikowskab, A., Jankowskia, K., Betlejewskab, K. and Hryniewiczb, W. 2001. Antibiotic susceptibility of bacterial strains isolated from urinary tract infections in Poland, Journal of Antimicrobial Chemotherapy, 47(6), 773–780.
Hyle, E.P., Lipworth, A.D., Zaoutis, T.E., Nachamkin, I., Fishman, N.O., Bilker, W.B., Mao, X. and Lautenbach, E. 2005. Risk Factors for Increasing Multidrug Resistance among Extended-Spectrum-Lactamase-Producing Escherichia coli and Klebsiella Species, Clinical Infectious Diseases, 40(9), 1317–1324.
Johnson, J.R. and Stell, A.L. 2000. Extended Virulence Genotypes of Escherichia coli Strains from Patients with Urosepsis in Relation to Phylogeny and Host Compromise, The Journal of Infectious Diseases, 181(1), 261–272.
Kaper, J.B., Nataro, J.P. and Mobley, H.L.T. 2004. Pathogenic Escherichia coli, Nature Reviews Microbiology, 2, 123–140.
Karimian, A., Momtaz, H. and Madani, M. 2012. Detection of uropathogenic Escherichia coli virulence factors in patients with urinary tract infections in Iran, African Journal of Microbiology Research, 6(39), 6811–6816.
Kollef, M.H., Sherman, G., Ward, S. and Fraser, V.J. 1999. Inadequate Antimicrobial Treatment of Infections: A Risk Factor for Hospital Mortality Among Critically Ill Patients, Chest, 115(2), 462–474.
Lautenbach, E., Strom, B.L., Bilker, W.B., Patel, J.B., Edelstein, P.H. and Fishman, N.O. 2002. Epidemiological Investigation of Fluoroquinolone Resistance in Infections Due to Extended‐Spectrum β‐Lactamase–Producing Escherichia coli and Klebsiella pneumoniae, Clinical Infectious Diseases, 33(8), 1288–1294.
Leboffe, M.J. and Pierce, B.E. (2011). Photographic Atlas for the microbiology laboratory, USA, Douglas N. Morton.
Litza, J. and Brill, J. 2010. Urinary tract infections, Prim Care, 37(3), 491–507.
M, N.S., Jayakumar, K. and Aravazhi, A.N. 2016. The effectiveness of antibiotics against a major uropathogen- Escherichia coli and its biofilm assay by phenotypic methods, International Journal of Research in Medical Sciences, 4(11), 4820–4828.
Manikandan, C. and Amsath, A. 2014. Antibiotic susceptibility pattern of Escherichia coli isolated from urine samples in Pattukkottai, Tamilnadu, International Journal of Current Microbiology and Applied Sciences, 3(10), 449–457.
Merza, N.S. 2013. Clonal Grouping of Uropathogenic Escherichia coli Using Different molecular Typing Methods in Kurdistan Region-Iraq, Ph.D Dissertation, University of Zakho. .
Merza, N.S., Fadhel Al Deen, I.S., Khalid, H.M. and Jubrael, J.M.S. 2016. Molecular Characterization of Extended Spectrum B-lactamase Producing Escherichia coli Isolated from Urine in Kurdistan Region-Iraq, International Journal of Chemical and Biomolecular Science, 2(2), 51–55.
Merza, N.S. and Jubrael, J.M.S. 2015. The Prevalence of Virulence Factors Among Uropathogenic Escherichia coli Strains Isolated From Different Hospitals in Kurdistan Region-Iraq, International Journal of Bioinformatics and Biomedical Engineering, 1(3), 338–343.
Miyazaki, J., Ba-Thein, W., Kumao, T., Obata Yasuoka, M., Akaza, H. and Hayshi, H. 2002. Type 1, P and S fimbriae, and afimbrial adhesin I are not essential for uropathogenic Escherichia coli to adhere to and invade bladder epithelial cells, FEMS Immunology and Medical Microbiology, 33(1), 23–26.
Mladin, C.; Usein, C.A.; Chifiriuc, M.C.; Palade, A.; Slavu, C. L.; Negut, M. and Damian, M. 2009. Genetic analysis of virulence and pathogenicity features of uropathogenic Escherichia coli isolated from patietns with neurogenic bladder, Romanian Biotechnological Letters, 14(6), 4900–4905.
Mukherjee, M., Basu, S., MuKherjee, S.K.M. and Majumder, M. 2013. Multidrug-resistance and extended spectrum beta-lactamase production in uropathogenic E. coli which were isolated from hospitalized patients in Kolkata, India, Journal of Clinical and Diagnostic Research, 7(3), 449–453.
Nicolle, L E. 2005. Complicated Urinary Tract Infection in Adults. Can J Infect Dis Med Microbiol, 16 (6), 349-60.
Nordstrom, L., Liu, C.M. and Price, L.B. 2013. Foodborne urinary tract infections: A new paradigm for antimicrobial-resistant foodborne illness, Frontiers in Microbiology, 4(29), 1–6.
Oliveira, F.A., Paludo, K.S., Arend, L.N.V.S., Farah, S.M.S.S., Pedrosa, F.O., Souza, E.M., Surek, M., Picheth, G. and Fadel-Picheth, C.M.T. 2011. Virulence characteristics and antimicrobial susceptibility of uropathogenic Escherichia coli strains, Genetics and Molecular Research, 10(4), 4114–4125.
Paterson, D.L. 2006. Resistance in gram-negative bacteria: Enterobacteriaceae, American Journal of Infection Control, 34 (5 SUPPL.), 20–28.
Poirel, L., Loo, M. Van De, Mammeri, H. and Nordmann, P. 2005. Association of Plasmid-Mediated Quinolone Resistance with Extended-Spectrum β-Lactamase VEB-1, Antimicrobial Agents and Chemotherapy, 49(7), 3091–3094.
Polse, R.F., Yousif, S.Y. and Assafi, M.S. 2016. Prevalence and Antimicrobial Susceptibility Patterns of Uropathogenic E. Coli among People in Zakho, Iraq. International Journal of Research in Medical Sciences, 4 (4), 1219–23.
Pourzare, M., Derakhshan, S. and Roshani, D. 2017. Distribution of Uropathogenic Virulence Genes in Escherichia coli Isolated from Children with Urinary Tract Infection in Sanandaj, Iran, Archives of Pediatric Infectious Diseases, 5(3), 1–6.
Rupp, M.E. and Fey, P.D. 2003. Extended Spectrum β-Lactamase (ESBL) Producing Enterobacteriaceae. Drugs, 63(4), 353–365.
Russo, T.A. and Johnson, J.R. 2000. Proposal for a New Inclusive Designation for Extraintestinal Pathogenic Isolates of Escherichia coli : ExPEC, The Journal of Infectious Diseases, 181(5), 1753–1754.
Sahm, D.F., Critchley, I.A., Kelly, L.J., Karlowsky, J.A., Mayfield, D.C., Thornsberry, C., Mauriz, Y.R. and Kahn, J. 2001. Evaluation of current activities of fluoroquinolones against gram-negative bacilli using centralized in vitro testing and electronic surveillance, Antimicrobial Agents and Chemotherapy, 45(1), 267–274.
Shah, A.A., Hasan, F., Ahmed, S. and Hameed, A. 2004. Extended-spectrum β-lactamases (ESBLs): Characterization, epidemiology and detection, Critical Reviews in Microbiology, 30(1), 25–32.
Sharma, N., Gupta, A., Walia, G. and Bakhshi, R. 2016. Pattern of antimicrobial resistance of Escherichia coli isolates from urinary tract infection patients: A three year retrospective study, Journal of Applied Pharmaceutical Science, 6(1), 062–065.
Šišková, P., Černohorská, L., Mahelová, M., Turková, K. and Woznicová, V. 2015. Phenotypes of Escherichia coli isolated from urine: Differences between extended-spectrum β-lactamase producers and sensitive strains, Journal of Microbiology, Immunology and Infection, 48(3), 329–334.
Siu, H.L., Reddy, S., Cheesbrough, J., Bolton, F.J., Willshaw, G., Cheasty, T., Fox, A.J. and Upton, M. 2008. Major uropathogenic Escherichia coli strain isolated in the Northwest of England identified by multilocus sequence typing, Journal of Clinical Microbiology, 46(3), 1076–1081.
Taghizadeh, G., Kulkarni, D.K., Delbari, A.S., and Taghizadeh, R. 2013. Antimicrobial susceptibility pattern of E. coli among patients attending at apadana hospital in ahwaz, Iran, International Journal of Research in Ayurveda and Pharmacy, 4(1), 120–122.
Tiwary, S.K., Sharan, S., Kumar, R. and Kumari, K. 2017. Antimicrobial susceptibility patterns of uropathogenic Escherichia coli and their prevalence among people in and around Dhanbad, Jharkhand, International Journal of Contemporary Medical Research, 4(8), 1637–1639.
Totsika, M., Moriel, D.G., Idris, A., Rogers, B.A., Wurpel, D.J., Phan, M.-D., Paterson, D.L. and Schembri, M.A. 2012. Uropathogenic Escherichia coli Mediated Urinary Tract Infection, Current Drug Targets, 13(11), 1386–1399.
Vieira, M.L.C., Santini, L., Diniz, A.L. and Munhoz, C. de F. 2016. Microsatellite markers: What they mean and why they are so useful, Genetics and Molecular Biology, 39(3), 312–328.
Wiles, T.J., Kulesus, R.R. and Mulvey, M.A. 2008. Origins and virulence mechanisms of uropathogenic Escherichia coli, Experimental and Molecular Pathology, 85(1), 11–19.
Zhanel, G.G., Hisanaga, T.L., Laing, N.M., DeCorby, M.R., Nichol, K.A., Weshnoweski, B., Johnson, J. ack, Noreddin, A., Low, D.E. and Karlowsky, J.A. 2006. Antibiotic resistance in Escherichia coli outpatient urinary isolates: final results from the North American Urinary Tract Infection Collaborative Alliance (NAUTICA), Int J Antimicrob Agents, 27(6), 468–76.
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