Detection of TEM, SHV and PER Type Extended-Spectrum ß-Lactamase Genes among Clinical Strains of Pseudomonas aeruginosa Isolated from Burnt Patients at Shafa-Hospital, Kerman, Iran

Document Type: Original Article

Authors

1 Department of Mirobiology, Kerman University of Medical Sciences, Kerman, Iran

2 Department of Bacteriology, Institute Pasteur of Iran, Tehran, Iran

Abstract

Objective(s)
This study was carried out to evaluate the existence of the TEM, SHV and PER ESBL genes in ESBL producing strains of Pseudomonas aeruginosa isolated from burnt patients at Shafa-hospital, Kerman, Iran.
Materials and Methods
A total of 120 strains of P. aeruginosa were isolated from 245 patients in burn unit of Shafa-hospital during January 2006 to December 2007. MIC of antibiotics was measured using agar dilution test. ESBL producing strains were detected by double-disc synergy method containing amoxicillin and amoxicillin+clavulanic acid and phenotypic confirmatory test. All the clinical isolates resistant to imipenem (IMP) were screened for  the production of MBL by E-test with IMP/IMP+EDTA strips. PCR and multiplex-PCR performed for the detection of different types of ESBL producing genes in ESBL positive isolates.
Results
Of 120 the isolates, 3-5% showed MIC greater than 16 µg/ml to IMP and meropenem, 66% showed MIC greater than 32 µg/ml to ceftazidime, 42% to azteronam and 60% of the isolates showed MIC greater than  64 µg/ml to cefotaxime, 41 (34%) confirmed as ESBL producers. Not any isolate could produce MBL (P≤0.05). The PCR assay of all ESBL producing isolates revealed that 6.6%, 4.1% and 2.5% of them were positive for SHV, PER and TEM genes, respectively.
Conclusion
Many ESBL producing strains of P. aeruginosa isolated from patients in burn unit of Shafa-hospital. However, none could produce MBL enzyme. The genes among ESBL producing strains were SHV, PER as well as TEM type of β-lacatamases. 

Keywords


1. Aggarwal R, Chaudhary U, Bala K. Detection of extended-spectrum β-lactamase in Pseudomonas aeruginosa. Indian J Pathol Microbiol 2008; 51:222-224.

2.  Nordmann P, Ronco E, Nass T. Characterzation of a novel extended-spectrum β-lactamase from Pseudomonas aeruginosa. Antimicrob Agents Chemother 1993; 37:962-769.

3.  Tredget E E, Shankowsky HA, Rennie R. Pseudomonas infections in the thermally injured patients. Burns 2004; 30:3-26.

4. Moniri R, Mosayebi Z, Movehdian A. Emergence of multi-drug-resistant Pseudomonas aeruginosa isolates in neonatal septicemia. Infec Dis Antimicrob Agents 2005; 22:39-44.

5. Brasme L, Nordmann P, Fidel F. Incidence of class-A extended -spectrum β-lactamases in Champagne-Ardenne (France): a 1 year prospective study. Antimicrob Chemother 2007; 60:956-964.

6.  Nathisuwan S, Burgess DS, Lewis JS II. Extended-spectrum β-lactamases: epidemiology, detection and treatment. Pharmaco 2001; 8:920-928.

7. Karmi Estahbanati H, Pour Kashani P, Ghanaatpisheh F. Frequency of Pseudomonas aeruginosa serotypes in burn wound infections and their resistance to antibiotics. Burns 2002; 28: 340-348.

8. Gehard FW, Poirel L, Nordmann P. Ambler calss-A extended-spectrum β-lactamases in Pesudomonas aeruginosa: Novel development and clinical impact. Antimicrob Agents Chemother 2003; 47:2385-2392.

9. Vahaboglu H, Oeztuerk R, Ayguen G. Widespread detection of PER-1-type extended- spectrum β-lactamases among nosocomial acinetobacter and Pseudomonas aeruginosa isolates in Turkey: A nation wide multicenter study. Animicrob Agents Chemother1997; 41:2265-2269.

10. Fluit A, Schmitz F. Class -I integron gene cassettes, mobility and epidemiology. Eur J Clincal Microbiol Infect Dis 1999; 18:761-770.

11. Poirel L, Rotimi V, Mokaddas E. VEB-1 like extended-spectrum β-lactamases in Pseudomonas aeruginosa, Kuwait. Emerg Infect Dis 2001; 7:486-470.

12. Lee S, Park Y, Kim M, Lee K. Prevalence of Ambler class -A and D β-lactamases among clinical isolates of Pseudomonas aerugnosa in Korea. Antimicrob Chemother 2005; 56:122-127.

13.Kim J, Lim Y, Rheem I. CTX-M and SHV-12 β-Lactamases are the most common extended-spectrum enzymes in clinical isolates of Escherichia coli and Kelabsiella pneumoniae collected from 3 university hospitals within Korea. FEMS Microbiol Letters 2005; 245:93-98.  

14.Song W, Lee K M, Kang H J. Microbiologic aspects of predominant bacteria isolated from the burn patients in Korea. Burns 2001; 2:136-139.

15. Shahcheraghi F, Feizabadi M M, Yamin V, Abiri R. Serovar determination, drug resistance patterns and plasmid profiles of Pseudomonas aeruginosa isolated from burn patients at two hospitals of Tehran, Iran. Burns 2003; 29:547-551

16. National Committee for Clinical Laboratory Standards (NCCLS): Performance of standards for antimicrobial disc susceptibility test-approved standard (document M2-A7) 7Th Ed. NCCLS, Pennsylvania, USA. (2000); ISBN 56-238208.

17. Norusis M Y. SPSS for windows: Base system user guide. Version 7.5- SPSS Inc., Chicago. 1993.

18. Mugnier P, Dubrous P, Casin I, Arlet G, et al. A TEM-derived extended spectrum β- lactamase in Pesudomonas aeruginosa.  Antimirob Agents Chemother1996; 40:2488-2493.

19. Song W, Lee KM, Kang HJ, et al. Microbiologic aspects of predominant bacteria isolated from the burn patients in Korea. Burns 2001; 2:136-139.

20.Khosravi A, Mihani F. Detection of Metallo-β-lactamase producing Pseudomonas aeruginosa strains isolated from burn patients in Ahwaz, Iran. Diagnost Microbiol Infectious Disease 2008; 6:125-128.

21.  Yamamano Y, Nishikawa T, Fujimura T, et al. Occúrance of PER-1 producing clinical isolates of Pseudomonas aeruginosa in Japan. J Antibiot  2006; 59:791-796.

22.  Jiang X, Zhang Z, Li M. Detection of extended-spectrum β-lactamases in clinical isolates of Pseudomonas aeruginosa. Antimicrob Agents Chemother 2006; 50:2990-2995.