Global Advanced Research Journal of Microbiology (GARJM) ISSN: 2315-5116
April 2018 Vol. 7(2): pp. 040-047
Copyright © 2018 Global Advanced Research Journals

 

Full Length Research Paper

 

Molecular Characterization of CTX-M ESBLs among Pathogenic Enterobacteriaceae isolated from different regions in Sudan

Hisham N Altayb1*, Mohamed A M Siddig2, Nagwa M El Amin3, Ahmed Ibrahim Hashim1, Maowia M. Mukhtar4

 

1College of medical laboratory science, Sudan University of science and technology, Sudan.

2Botany department, Faculty of Science, University of Khartoum, Sudan

3Department of microbiology, Faculty of medicine, university of Khartoum, Sudan

4Institute of Endemic Diseases, University of Khartoum, Khartoum, Sudan

Email: hishamaltayb@yahoo.com

 

Accepted 07 April, 2018

 

Abstract

This is a cross sectional study conducted to detect and characterize CTX-M genes among extended spectrum β-lactamases (ESBLs) producing Enterobacteriaceae isolated from different regions in Sudan.  A total of 305 of clinical Enterobacteriaceae sp. isolates were collected from different regions in Sudan. ESBLs production was initially screened by cefotaxime, cefepime and ceftazidime, and then confirmed by disk combination method and PCR. DNA sequencing was done to differentiate between  blaCTX-M genotypes. Escherichia coli was the most predominant (58%) isolate, followed by Klebsiella pneumoniae (26.6%), Citrobacter freundii (3.6%), Enterobacter species (6.2%) and Proteus species (5.6%). ESBLs were detected by disk combination method in 128 (42%) isolates; Khartoum state 64% (23/36), Gizera state 54% (54/100), Sinnar state 53% (49/92) and White Nile state 2.6% (2/77). Three quarters of the ESBLs producers (96/128) were positive for blaCTX-M genes by PCR. blaCTX-M-15 was the most reported one 78.3% (18/23), followed by blaCTX-M-14 13% (3/23), blaCTX-M-27 4.3% (1/23) and blaCTX-M-98 4.3% (1/23). There was a transition mutation (substitution of A with G at position 25) in the blaCTX-M gene (ID: KP309815), that affect protein structure. We concluded that blaCTX-M-15 was the most commonly encountered genes and widely spread in different Sudanese regions. 

Keywords:  blaCTX-M; Beta-Lactamases Genes; Enterobacteriaceae; Sudan.

 

 

REFERENCES

 

Abdelmoneim A, Idris E, Lloyd M, Lukman T (2005). Self-medication with Antibiotics and Anti malarials in the community of Khartoum State, Sudan. J Pharm Pharmaceut Sc. 2005;8(2):326-331.

Alsadig G, Mohamed A, Arbab, Sawsan AH. Aldeaf (2014). Allele Frequency of P53 Gene Arg72Pro In Sudanese Meningioma Patients And Controls. In. J. of SCIENTIFIC & TECHNOLOGY RES. 2014; 3 (6). 2014; 2277-8616.

An e-Science approach with life scientist friendly interfaces (2010). “Protein structure analysis of mutations causing inheritable diseases”   BMC Bioinformatics. 8; 11(1):548; PMID: 21059217; DOI: 10.1186/1471-2105-11-548. Sites in protein structures”. J Mol Biol: 2010; 339(3):607-33.

Atschul SF, Madden TL, Schaffer AA (1997). Gapped BLAST and PSI-BLAST. A new generation of protein database search programmes”. NucleicAcid Res 25: 3389-3402.

Bonnet R (2004). Growing group of extended-spectrum β-lactamases: the CTX-M enzymes. Antimicrob. Agents Chemother. 2004;48, 1-14.

Cantón R (2008). “Epidemiology and evolution of β-lactamases,”in Evolutionary Biology of Bacterial and Fungal Pathogens, eds F.Baquero, C. ombela, G.H. Casslel, and J. A. Gutierrez-Fuentes (Washington: ASM Press). 2008;249–270.

Cao V, Lambert T,  Courvalin P (2002). ColE1-like plasmid pIP843 of Klebsiella pneumoniae encoding extended-spectrum ß-lactamases CTX-M-17. Antimicrob Agents Chemother. 2002;46, 1212–1217.

Clinical and Laboratory Standards Institute (2011). Performance Standards for Antimicrobial   Susceptibility Testing; Twenty First informational Supplement, CLSI document M 100-S21. Wayne, PA; USA: Clin and Lab standards Institute; 2011.

Gröbner S, Linke D, Schutz W, Fladerer C, Madlung J (2009).  Emergence of carbapenem-non-susceptible extended-spectrum β-lactamase-producing Klebsiella pneumoniae isolates at the university hospital of Tübingen, Germany. J Med Microbiol 2009, 58(Pt 7): 912–922.

Hall TA (1999). “BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT”. Nucl. Acids. Symp. Ser. 1999;41:95-98.

Hamedelnil FY, Eltayeb HN (2012).  Molecular detection of Extended Spectrum β-lactamases (ESBLs) genes in E. coli isolated from urine specimens. Int J of advanced and scientific research. October, 2012, ISSN 2249-9954.

Hawkey PM, Jones AM (2009).  The changing epidemiology of resistance. J. Antimicrob. Chemother. 2009; 64, i3–i10.

IBM SPSS Statistics for Windows, Version 21.0 (2012). Armonk, NY: IBM Corp, Released 2012. www.i bm.com/software/analytics/spss/ IBM Corp.

John B, Alexandre L, Mark B (2001). Gene Mark S: a self-training method for prediction of gene starts in microbial genomes. Implications for finding sequence motifs in regulatory regions”.  Nucleic Acids Research 29. 2001; 2607-2618.

Karim A, Poirel L, Nagarajan S, Nordmann P (2001). Plasmid-mediated extended-spectrum β-lactamase (CTX-M-3 like) from India and gene association with insertion sequence ISEcp1. FEMS Microbiol Lett 2001, 201(2): 237-241.

Livermore DM, Hawkey PM (2005). CTX M changing the face of ESBLs in UK. J Antimicrob Chemother. 2005; 56:451–4.

Livermore DM, Woodford. The β-lactamase threat in Enterobacteriaceae, Pseudomonas and Acinetobacter. Trends Microbiol 2006, 14(9): 412-420.

Mekki AH, Hassan AN, Elsayed DM (2010). Extended Spectrum Beta Lactamases Among Multi Drug Resistant Escherichia Coli and Klebsiella Species Causing Urinary Tract Infections In Khartoum. J Bact Res.2010; 2(3):18-21.

Mobaiyen H, Nahaei MR, Amirmozafari N, Moaddab SR, Hoseni F (2007). First report of CTX-M type β -lactamase producing clinical Escherichia coli isolates from intensive care units in Tabriz hospitals. Abstr. 1st Iranian Cong of clinical Microbio, Shiraz, Iran, Abstracts 'Book, 2007; 152.

Munday CJ, Whitehead GM, Todd NJ (2004). J Antimicrob Chemother. 2004; 54:628–33.

Murray PR, Baron EJ, Jorgensen JH, Landry ML, Michael (2007). Manual of Clinical microbiology. 9th ed. Washington: American Society for Microbiology; 2007.

Omar BA, Alfadel OO, Atif HA,  Mogahid ME (2013). Prevalence of TEM, SHV and CTX-M genes in Escherichia coli and Klebsiella spp. in urinary isolates from Sudan with confirmed ESBL phenotype, Life Science Journal 2013;10(2) http://www.lifesciencesite.com.

Paterson DL, Bonomo RA (2005). Extended-spectrum beta-lactamases a clinical update. Clin Microbiol Rev 2005, 18(4):657-686.

Pena CM, Pujol A, Ricart C, Ardanuy J, Ayats (1997). Risk factors for faecal carriage of Klebsiella pneumoniae producing extended spectrum beta-lactamase (ESBL-KP) in the intensive care unit. J. Hosp. Infect. 1997;35:9–16.

Poirel L, Kampfer P, Nordmann P (2002). Chromosome-encoded Ambler class A beta-lactamase of Kluyvera georgiana, a probable progenitor of a subgroup of CTX-M extended-spectrum beta-lactamases. Antimicrob Agents Chemother. 2002;46(12):4038-40.

Quinteros M, Radice M, Gardella N, Rodriguez MM, Costa N, Korbenfeld D (2003).  Extended-spectrum Beta lactamases in Enterobacteriaceae in Buenos Aires, Argentina, public hospitals. Antimicrob Agents Chemother 2003; 47: 2864-7.

Schwaber MJ, Carmeli Y (2007). Mortality and delay in effective therapy associated with extended-spectrum beta-lactamase production in Enterobacteriaceae bacteraemia: a systematic review and meta-analysis. Journal of Antimicrobial Chemotherapy 2007; 60(5):913-20.

Wayne, Pa, USA. Performance standards for antimicrobial susceptibility testing, eighteenth informational supplement,” Tech. Rep. M100-S18, Clinical and Laboratory Standards Institute; 2008.

Wayne, PA. Clinical and Laboratory Standards Institute (2006). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically: approved standard. 7th ed. M7–A7. Clinical and Laboratory Standards Institute; 2006.

 

This is a cross sectional study conducted to detect and characterize CTX-M genes among extended spectrum β-lactamases (ESBLs) producing Enterobacteriaceae isolated from different regions in Sudan.  A total of 305 of clinical Enterobacteriaceae sp. isolates were collected from different regions in Sudan. ESBLs production was initially screened by cefotaxime, cefepime and ceftazidime, and then confirmed by disk combination method and PCR. DNA sequencing was done to differentiate between  blaCTX-M genotypes. Escherichia coli was the most predominant (58%) isolate, followed by Klebsiella pneumoniae (26.6%), Citrobacter freundii (3.6%), Enterobacter species (6.2%) and Proteus species (5.6%). ESBLs were detected by disk combination method in 128 (42%) isolates; Khartoum state 64% (23/36), Gizera state 54% (54/100), Sinnar state 53% (49/92) and White Nile state 2.6% (2/77). Three quarters of the ESBLs producers (96/128) were positive for blaCTX-M genes by PCR. blaCTX-M-15 was the most reported one 78.3% (18/23), followed by blaCTX-M-14 13% (3/23), blaCTX-M-27 4.3% (1/23) and blaCTX-M-98 4.3% (1/23). There was a transition mutation (substitution of A with G at position 25) in the blaCTX-M gene (ID: KP309815), that affect protein structure. We concluded that blaCTX-M-15 was the most commonly encountered genes and widely spread in different Sudanese regions.

Current Issue

Viewing Options

View Full Article - PDF
Download Full Article - PDF

Search for Articles

Hisham N Altayb on Google Scholar
Hisham N Altayb on Pubmed
Mohamed A M Siddig on Google Scholar
Mohamed A M Siddig on Pubmed
Nagwa M El Amin on Google Scholar
Nagwa M El Amin on Pubmed
Ahmed Ibrahim Hashim on Google Scholar
Ahmed Ibrahim Hashim on Pubmed
Maowia M. Mukhtar on Google Scholar
Maowia M. Mukhtar on Pubmed

Statistics

Viewed 243
Printed 119
Downloaded 230
Powered By iPortal Works