Prevalence and Molecular Characterization of Plasmid-mediated Extended-Spectrum β-Lactamase Genes (balaTEM, blaCTX and blASHV) Among Urinary Escherichia coli Clinical Isolates in Mashhad, Iran

Document Type: Original Article

Authors

1 Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran

2 Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran

Abstract

Objective(s)
Extended-spectrum beta-lactamase (ESBL) producing bacteria have an important role in nosocomial infections. Due to the limited availability of information about the molecular epidemiology of ESBL producing bacteria in Mashhad, we decided to investigate about TEM, CTX and SHV ESBLs among urinary Escherichia coli isolates in Mashhad, a city in northeast Iran.
Materials and Methods                        
One hundred and eleven clinical isolates of E. coli were diagnosed from hospitalized patients in 2009. After performing antibiogram and phenotypic confirmation test, polymerase chain reaction (PCR) was performed by blaTEM, blaSHV and blaCTX primers and restriction digestion was carried out using PstI and TaqI (Fermentas-Lithuania) for confirmation.
Results
ESBL producers of E. coli isolates were 33.3%. Among 37 ESBL-producing isolates, 35 (94.6%), 21 (56.8%) and 5 (13.5%) were shown to have blaCTX, blaTEM and blaSHV, genes respectively. Co-resistance to non-beta lactam antibiotics was observed more with ESBL producers (P < 0.05).
Conclusion
The results showed that the studied ESBL genes are found with high prevalence and among them blaCTX is more widespread in urine E. coli isolates in Mashhad.
 

Keywords


1. Sanders CC, Jr Sanders WE. Beta-lactamase in gram-negative bacteria: global trends and clinical impact. Clin Infect Dis 1992; 15:824-839.

2. Paterson DL, Bonoma RA. Extended-spectrum β-lactamases: a clinical update. Clin Microbiol Rev 2005; 18:657-686.

3. Branger C, Zamfir O, Geoffroy S, Laurans G, Arlet G, Thien HV, et al. Genetic background of Escherichia coli and extended- spectrum β-lactamases type. Emerg Infect Dis 2005; 11:54-61.

4. Bradford PA. Extended spectrum beta-lactamases in the 21st century: characterization, epidemiology and the detection of this important resistance threat. Clin Microbiol Rev 2001; 14:933-951.

5. Prescott LM, Harley JP, Klein DA. Microbiology. 5th ed. USA: Mc Graw Hill; 2002.

6. Peleg AY, Hooper DC. Hospital-acquired infections due to Gram-negative bacteria. N Engl J Med 2010; 362:1804-1813.

7. Livemore DM. Beta-lactamases in laboratory and clinical resistance. Clin Microbiol Rev 1995; 8:557-584.

8. Prinarakis EE, Miriagou V, Tzelepi E, Gazouli M., Tzouvelekis LS. Emergence of an inhibitor-resistant beta-lactamases (SHV-10) derived from an SHV-5 variant. Antimicrob Agents Chemother 1997; 41:838-840.

9. Tzouvelekis LS, Tzelepi E, Tassios PT, Legakis NJ. CTX-M-type beta-lactamases: an emerging group of extended spectrum enzymes. Int J Antimicrob Agents 2000; 14:137-142.

10. Bradford PA,Yang Y, Sahm D, Grope I, Gardovska D, Storch G. CTX-M-5, a novel cefotaxime-hydrolyzing beta-lactamase from an outbreak of S. typhimurium in Latvia. Antimicrob Agents Cehmother 1998; 42:1980-1984.

11. Nathisuwan S, Burgess DS, Lewis JS. ESBLs: epidemiology, detection and treatment. Pharmacotherapy 2007; 21:920-928.

12. National Committee for Clinical Laboratory Standards, Performance standards for antimicrobial susceptibility testing, 12th informational supplement. M100-S12, National Committee for Clinical Laboratory Standards. Wayne: PA. 2002.

13. Jarlier V, Nicolas MH, Fournier G, Philippon A. Extended-spectrum beta-lactamases conferring transferable resistance to newer beta-lactam agents in Enterobacteriaceae: hospital prevalence and susceptibility patterns. Rev Infect Dis 1988; 10:867-878.

14. M'Zali FH, Chanawong A, Kerr G, Birkenhead D, Hawkey PM. Detection of extended- spectrum beta-lactamases in members of the family Enterobacteriaceae: comparison of the MAST DD test, the double disk and the E test ESBL. Antimicrob Agents Chemother 2000; 45:881-885.

15. National Committee for Clinical Laboratory Standards. Performance standards for antimicrobial disk susceptibility test, 7th ed. Approved standards, NCCLS Doucument M2-A7, Vol; 20, no.1; Wayne, PA. 2000.

16. Sundsfjord A, Simonsen GS, Haldorsen BC, Haaheim H, Hjelmevoll SO, Littauer P, et al. Genetic methods for detection of antimicrobial resistance. APMIS  2004; 112:815-837.

17. Pagani L, Dell' Amico E, Migliavacca R, D'Andrea MM, Giacobone E, Amicosante G, et al. Multiple CTX-M-type extended-spectrum beta-lactamases in nosocomial isolates of Enterobacteriaceae from a hospital in northern Italy. J Clin Microbiol 2003; 41:4264-4269.

18. Machado E, Coque TM, Canton R, Novais A, Sousa JC, Baquero F, et al. High diversity of extended- spectrum β-lactamases among clinical isolates of Enterobacteriaceae from Portugal. J Antimicrob Chemether 2007; 60:1370-1374.

19. Al-Zarouni M, Senok A, Rashid F, Al-Jesmi SM, Panigrahi D. Prevalence and antimicrobial susceptibility pattern of extended-spectrum beta-lactamase-producing Enterobacteriaceae in the United Arab Emirates. Med Princ Pract 2008; 17:32–36.

20. Malhotra VL, Khandpur N, Dass A, Mehta G. Prevalence of extended spectrum beta-lactamases producing clinical isolates from patients of urinary tract infection in a tertiary care hospital in Dehli. J Commun Dis 2008; 40:269-272.    

21. Babypadmini S, Appalaraju B. Extended spectrum beta-lactamases in urinary isolates of Escherichia coli and Klebsiella  pneumoniae- Prevalence and susceptibility patients in a tertiary care Hospital. Ind J Med Microbiol 2004; 22:172-174.

22. Yagi T, Krukawa H, Shibata N, Shibayama K, Arkawa Y. A preliminary survey of extended spectrum β-lactamases in clinical isolated of K. pneumonia and E. coli in Japan. FEMS Microbiol 2000; 184:53-56.

23. Lewis MT, Yamaguchi K, Biedenbach DJ, Jones RN. In vitro evaluation of cefepime and other broad-spectrum beta-lactams in 22 medical centers in Japan: a phase II trial comparing two annual organism samples. The Japan Antimicrobial Resistance Study Group. Diagn Microbiol Infect Dis 1999; 35:307-315.

24. Stobberingh EE, Arends J, Hoggkamp-Korstanje JAA, Goessens WH, Visser MR, Buiting AG, et al. Occurrence of extended-spectrum β-lactamases in Dutch hospitals. Infection 1999; 27:348-354.

25. Quinn JP, Bradford PA, Goering RV, Nathan C, Bush K, Weinstein RA. Multiple antibiotic- resistant Klebsiella and Escherichia coli in nursing homes. J Am Med Assoc 1999; 281:517-523.

26. Ndugulile F, Jureen R, Harthuy S, Urassa W, Langeland N. Extended spectrum beta-lactamases among Gram-negative bacteria of nosocomial origin from an intensive care unit of a tertiary health facility in Tanzania. BMC Infect Dis 2005; 5:86.

27. Goyal A, Prasad KN, Prasad A, Gupta S, Ghoshal U, Ayyagari A. Extended spectrum beta-lactamases in Escherichia coli and Klebsiella  pneumoniae and associated risk factors. Indian J Med Res 2009; 129:695-700.

28. Mendonca N, Leitaco J, Manageiro V, Ferreira E, Canica M. Spread of ESBL CTX-M producing E. coli clinical isolates in community and nosocomial environments in Purtugal. Antimicrob Agents Chemother 2007; 51:1946-1955.

29. Lavigne JP, Marchandin H, Delmas J, Moreau J, Bouziges N, Lecaillon E, et al. CTX-M beta-lactamases-producing Escherichia coli in French Hospitals: prevalence molecular epidemiology and risk factors. J Clin Microbiol 2007; 45:620-626.

30. Fang H, Ataker F, Hedin G, Dornbusch K. Molecular epidemiology of ESBLs among Escherichia coli isolates collected in a Swedish Hospital and its associated health care facilities from 2001 to 2006. J Clin Microbiol 2008; 46:707-712.