International Journal of Chemical and Biomolecular Science
Articles Information
International Journal of Chemical and Biomolecular Science, Vol.2, No.2, Apr. 2016, Pub. Date: Apr. 5, 2016
Molecular Characterization of Extended Spectrum B-lactamase Producing Escherichia coli Isolated from urine in Kurdistan Region-Iraq
Pages: 51-55 Views: 2370 Downloads: 1481
Authors
[01] Narmin S. Merza, Scientific Research Center Duhok University, Duhok, Iraq.
[02] Iman H. Fadhel Al Deen, Scientific Research Center Duhok University, Duhok, Iraq.
[03] Haval M. Khalid, Biology Department, College of Science, Zakho University, Duhok, Iraq.
[04] Zaynab H., Scientific Research Center Duhok University, Duhok, Iraq.
[05] Jaladet M. S. Jubrael, Scientific Research Center Duhok University, Duhok, Iraq.
Abstract
Molecular characterization of ESBL-related bla genes including blaTEM, blaSHV, and blaCTX-M has been performed for Escherichia coli isolated from urine and collected from three cities in Kurdistan/region/Iraq (Erbil, Sulymani and Duhok). One hundred sixty nine isolates of E. coli have been identified and their production of ESBLs enzymes have been determined using phenotypic methods. All these isolates were successfully amplified producing a single band of the uidA locus in all strains with a molecular weight of about 670bp in order to confirm at molecular level that all these isolates were E. coli. One hundred sixty ESBL E. coli isolates out of 169 appeared to have one or more ESBLs genes accounting for 94.7%. CTX-M constituted the high prevalent type of ESBLs genes compared to the others represented by 94.1% of all isolates in all the three cities of Kurdistan region followed by TEM and SHV in a percentages of 43.8% and 2.5%, respectively. In Duhok, TEM showed the higher prevalence (60.8%) in comparison to the other two cities in percentages of 36.2% for Sulaimania while Erbil represented by 25%. Furthermore, it was clear that SHV type of ESBLs had the lower prevalence of all types and there were only four isolates out of 160 appeared to carry this type of gene representing 2.5%. The presence and/or absence of the three genes in all isolates were also investigated and it was shown that 86/160 isolates (53.75%) had the CTX-M gene only while the rest of genes were lacking. Moreover, 69/160 isolates had both CTXM and TEM. Interestingly, 3/160 harbored all three involved genes. The isolates characterized by the presence only TEM gene and those that had both CTX-M and SHV, shared the same percentage (0.6%). after taking sequencing of the PCR product of studied genes for 12 E coli isolates into consideration, it was obvious that all the PCR products of CTX-M were belonged to type CTXM-15; while TEM-1 type appeared predominant among all sequences PCR product for TEM gene.. Finally, from the three isolates which revealed positive PCR amplification for the SHV gene, two isolates showed 100% similarity to the SHV-12 genome type while the rest single isolate was similar (99%) to SHV11.
Keywords
PCR Assay, ESBLs, Escherichia coli
References
[01] Adamus-Bialek, W.; Wojtasik, A.; Majchrzak, M.; Sosnowski, M., and Parniewski, P. (2009). (CGG) 4-Based PCR as a Novel Tool for Discrimination of Uropathogenic Escherichia coli Strains: Comparison with Enterobacterial Repetitive Intergenic Consensus-PCR. Journal of Clinical Microbiology, 47(12): 3937–3944.
[02] Aktaş Z, Gönüllü N, Kayacan ÇB, Özdem A, Carattoli A, Yong DE, Walsh T. (2009). CTX-M-15 Carried On Incn-Type Plasmids in Klebsiella pneumoniae. Türkiye Klinikleri J Med Sci; 29.
[03] Al-Mizory S. (2007). Study of the Bacterium Pseudomonas aeruginosa Isolated from Different Sources at Dohuk City. M. Sc. Thesis, Collage of Education, Dohuk University.
[04] Bali EB, Acik L, Sultan N. (2010) Phenotypic and molecular characterization of SHV, TEM, CTX-M and extended-spectrum β-lactamase produced by Escherichia coli, Acinetobacter baumannii and Klebsiella isolates in a Turkish hospital. African J Microb Res 2010; 4(8): 650-654.
[05] Bonnet R (2004) Growing group of extended-spectrum beta-lactamases: the CTX-M enzymes. Antimicrob Agents Chemother 48: 1-14.
[06] Canton R, Coque TM (2006) The CTX-M beta-lactamase pandemic. Curr Opin Microbiol; 9: 466-475.
[07] Gonullu Nevriye,1 Aktas Z., Kayacan C. Bal, Salcioglu M. et al., (2008) Dissemination of CTX-M-15 -Lactamase Genes Carried on Inc FI and FII Plasmids among Clinical Isolates of Escherichia coli in a University Hospital in Istanbul, Turkey; Journal of Clinical Microbiology, vol46, No. 3
[08] Grover S. S, Sharma M, Chattopadhya D, Kapoor H, Pasha ST, Singh G. Phenotypic and genotypic detection of ESBL mediated cephalosporins resistance in Klebsiella pneumoniae: Emergence of high resistance against cefepime, the fourth generation cephalosporins. J Infect 2006; 54: 279-88.
[09] Harada Yosuke, Morinaga Yoshitomo, Yamada Koichi, Migiyama Yohei, Nagaoka Kentaro, Uno Naoki, et al., (2013) Clinical and Molecular Epidemiology of Extended-Spectrum β-lactamase- Producing Klebsiella pneumoniae and Escherichia coli in a Japanese Tertiary Hospital, J Med Microb Diagn, 2:3.
[10] Jazi MF, F Valehi, Talebi A, Rastegar Lari A.(2007) Molecular characterization of resistance to Extended - Spectrum antibiotics in Escherichia coli and Klebsiella pneumoniae. Iran J Med Microbiol 2007; 2:27-34.
[11] Kaftandzieva A, Trajkovska-Dokic E, Panovski N (2011) Prevalence and molecular characterization of Extended spectrum beta lactamase producing Escherichia coli and Klebsilla pneumonia. Contributions, Sec. Biol. Med. Sci., MASA, XXXII, 2, p. 129–141.
[12] Khalaf Noha G, Eletreby Mona M, and Hanson Nancy D (2009) Characterization of CTX-M ESBLs in Enterobacter cloacae, Escherichia coli and Klebsiella pneumoniae clinical isolates from Cairo, Egypt, BMC Infect Dis.; 9: 84.
[13] Khalid H. M. (2013). Molecular Epidemiology of Extended-Spectrum β-Lactamase Producing Klebsiella pneumoniae in Kurdistan Region /Iraq, PhD thesis submitted to Faculty of Science-University of Zakho-Iraq.
[14] Kiratisin P, Apisarnthanarak A, Laesripa C, Saifon P. (2008) Molecular characterization and epidemiology of extended-spectrum-beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolates causing health care-associated infection in Thailand, where the CTX-M family is endemic; Antimicrob Agents Chemother; 52(8): 2818-24.
[15] Mahboobeh Nakhaei Moghaddam, Mehrdad Hashemi Beidokhti, Saeid Amel Jamehdar, and Martha Ghahraman (2014) Genetic properties of blaCTX-M and blaPER β-lactamase genes in clinical isolates of Enterobacteriaceae by polymerase chain reaction, Iran J Basic Med Sci.; 17(5): 378–383.
[16] Moosavian M. and Deiham B. (2012). Distribution of TEM, SHV and CTX-M Genes among ESBL-producing Enterobacteriaceae isolates in Iran; African Journal of Microbiology Research Vol. 6(26), pp. 5433-5439.
[17] Natarajan Vidhya and Singaram S. Sivasubramanian (2013) Prevelance of bla TEM gene in uropathogenic Escherichia coli isolated from tertiary care hospitals in Coimbator, South India. International Journal of Pharmacy and Pharmaceutical Sciences, Vol 5, Suppl 3.
[18] Nazik Hasan, Bektöre Bayhan, Öngen Betigül, İlktaç Mehmet, Özyurt Mustafa, Kuvat Nuray; et al, (2011) Plasmid-Mediated Quinolone Resistance Genes in Escherichia coli Urinary Isolates from Two Teaching Hospitals in Turkey: Coexistence of TEM, SHV, CTX-M and VEB-1 Type β-lactamases. Tropical Journal of Pharmaceutical Research, 10(3): 325-333.
[19] Paterson DL1, Bonomo RA (200) Extended-spectrum beta-lactamases: a clinical update; Clin Microbiol Rev. 2005 Oct; 18(4): 657-86.
[20] Pfaller Michael A. and Segreti John (2006) Overview of the Epidemiological Profile and Laboratory Detection of Extended-Spectrum β-Lactamases, Clinical Infectious Diseases; 42: S153–63.
[21] Pitout Johann DD (2009) Recent changes in the epidemiology and management of extended-spectrum b-lactamase-producing Enterobacteriaceae, F1000 Medicine Reports, 1: 84.
[22] Rupp Mark E. and Fey Paul D. (2003) Extended Spectrum B-Lactamase (ESBL)-Producing Enterobacteriaceae Considerations for Diagnosis, Prevention and Drug Treatment, Drugs; 63 (4): 353-365.
[23] Salah M. A. Abu 1, Badran E. F., Shehabi A A.(2013) High incidence of multidrug resistant Escherichia coli producing CTX-M-type ESBLs colonizing the intestine of Jordanian; International Arabic Journal of antimicrobial agents; Vol. 3 No. 4: 3.
[24] Sharma J, Sharma M, Ray P. (2010) Detection of TEM and SHV genes in Escherichia coli & Klebsiella pneumoniae isolates in a tertiary care hospital from India. Indian J Med Res; 132: 332-336.
[25] Shiri NV, Orly HM, David S, Dan T, Boris K, Yehuda C.(2003) Occurrence and phenotypic characteristics of extended-spectrum β-lactamases among members of family Enterobacteriaceae at the Tel-Aviv Medical Center (Israel) and evaluation of diagnostic tests. J Clin Microbiol; 41: 155-8.
[26] Thenmozhi S., Moorthy K., Sureshkumar B. T., Suresh M.(2014) Antibiotic Resistance Mechanism of ESBL Producing Enterobacteriaceae in Clinical Field: A Review, Int. J. Pure App. Biosci. 2 (3): 207-226
[27] Vidhya N. and Sudha S. S (2013) prevalence of bla CTX-M extended spectrum beta lactamase gene in uropathogenic Escherichia coli. World Journal of Pharmacy and Pharmaceutical Sciences, Volume 2, Issue 6, 6548-6558.
[28] Yumuk Z, Afacan G, Nicolas-Chanoine MH, Sotto A, Lavigne JP. (2008) Turkey: a further country concerned by community-acquired Escherichia coli clone O25-ST131 producing CTX-M-15. J Antimicrob Chemother; 62: 284-288.
[29] Yun-Tae, K., K. Tae-Un and B. Hyung-Suk, (2006). Characterization of extended spectrum β- lactamase genotype TEM, SHV and CTX-M producing Klebsiella pneumoniae isolated from clinical specimens in Korea. J. Microbiol. Biotechnol., 16: 889-95.
[30] Zahar JR, E Bille, D Schnell, F Lanternier, Mechai F, Masse V, Nassif X, Lortholary O. (2009) Extension of beta-lactamases producing bacteria is a worldwide concern. Med Sci (Paris); 5(11): 939-944.
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