Document Type : Original Article
Authors
- Davood Kalantar-Neyestanaki Kalantar-Neyestanaki 1
- Fereshteh Jabalameli 2
- Akbar Mirsalehian 3
- Mohammad Emaneini 4
1 Assistant Professor, Department of Microbiology, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran
2 Assistant Professor, Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
3 Professor, Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
4 Associate Professor, Department of Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
Abstract
Background & Aims: The production of extended-spectrum-β-lactamases (ESBLs) is the main mechanism of resistance to β-lactam antibiotics. The outbreak of isolates simultaneously possessing several resistance mechanisms to β-lactam antibiotics caused a decrease in sensitivity of the confirmatory tests for ESBL. The aim of this study was the evaluation of the β-lactamase disk test method in the detection of ESBLs in clinical isolates of multidrug-resistant Pseudomonas aeruginosa.
Methods: A total of 100 multidrug-resistant P. aeruginosa isolates were recovered from burn patients. The sensitivity of the isolates to different antibiotics was determined using the standard disk diffusion method. ESBL-producing isolates were detected through the combination disk test with clavulanic acid, double disk synergy test, and β-lactamase disk test. Carbapenemase-producing isolates were detected using the Modified Hodge Test (MHT). The ESBLs genes (blaTEM, blaOXA, blaPER, blaSHV, blaCTX-M and blaPSE) were determined through polymerase chain reaction (PCR).
Results: All isolates were multidrug resistant. Only 3 isolates were detected as ESBL-producing isolates through combination disk test. No ESBL-producing isolates were detected through double disk synergy test. Among the 100 studied isolates, 87% were detected as ESBL-producing isolates and 68% as carbpenemase-producing isolates through β-lactamase disk test. . The prevalence of bla
TEM
, bla
OXA
, and bla
PER
among isolates were 97%, 61%, and 13%, respectively. All isolates were negative for bla
SHV
, bla
PSE
, and bla
CTX-M
.
Conclusion: According to the results obtained in this study, the β-lactamase disk test is suitable for the detection of ESBLs in multidrug resistant isolates. However, further investigation is required.
Keywords
- Pfeifer Y, Cullik A, Witte W. Resistance to cephalosporins and carbapenems in Gram-negative bacterial pathogens. Int J Med Microbiol 2010; 300(6): 371-9.
- Poole K. Pseudomonas aeruginosa: resistance to the max. Front Microbiol 2011; 2: 65.
- Bush K, Jacoby GA. Updated functional classification of beta-lactamases. Antimicrob Agents Chemother 2010; 54(3): 969-76.
- Falagas ME, Karageorgopoulos DE. Extended-spectrum beta-lactamase-producing organisms. J Hosp Infect 2009; 73(4): 345-54.
- Sundin D. Hidden beta-lactamases in the enterobacteriaceae – dropping the extra disks for detection, part II. Clinical Microbiology Newsletter 2009; 31(7): 47-52.
- Willems E, Verhaegen J, Magerman K, Nys S, Cartuyvels R. Towards a phenotypic screening strategy for emerging beta-lactamases in Gram-negative bacilli. Int J Antimicrob Agents 2013; 41(2): 99-109.
- Doi Y, Paterson DL. Detection of plasmid-mediated class C beta-lactamases. Int J Infect Dis 2007; 11(3): 191-7.
- Jacoby GA. AmpC beta-lactamases. Clin Microbiol Rev 2009; 22(1): 161-82, Table.
- Mahon CR, Lehman D, Manuselis G. Textbook of diagnostic microbiology. 3rd ed. Philadelphia, PA: Saunders Elsevier; 2007.
- Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; twenty-second informational supplement [Online]. [cited 2012 Jan]; Available from: URL: http://antimicrobianos.com.ar/ATB/wp-content/uploads/2012/11/M100S22E. p. 58, 60, 114-22.
- Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; twenty-fourth informational supplement [Online]. [cited Jun 2014]; Available from: URL: http://ncipd.org/control/images/NCIPD_docs/CLSI_M100-S24.pdf
- Neyestanaki DK, Mirsalehian A, Rezagholizadeh F, Jabalameli F, Taherikalani M, Emaneini M. Determination of extended spectrum beta-lactamases, metallo-beta-lactamases and AmpC-beta-lactamases among carbapenem resistant Pseudomonas aeruginosa isolated from burn patients. Burns 2014; 40(8): 1556-61.
- Martine E, Escobar Pérez J, Márquez C, Vilacoba E, Centró D, Leal AL, et al. Emerging and existing mechanisms co-operate in generating diverse ß-lactam resistance phenotypes in geographically dispersed and genetically disparate Pseudomonas aeruginosa strains. Journal of Global Antimicrobial Resistance 2013; 1(3): 135-42.
- Poole K. Resistance to beta-lactam antibiotics. Cell Mol Life Sci 2004; 61(17): 2200-23.
- Jacoby GA, Munoz-Price LS. The new ß-Lactamases. N Engl J Med 2005; 352(4): 380-91.
- Kalantar D, Jabalameli F, Emaneini M. The modified Hodge test for identification of Klebsiella pneumoniae carbapenemase producing isolates. Burns 2013; 39(2): 370-1.
- Kalantar D, Mansouri S. Emergence of multiple β-lactamases produced by Escherichia coli clinicalisolates from hospitalized patient in Kerman, Iran. Jundishapur J Microbiol 2010; 3(4): 137-45.
- Mansouri S, Kalantar D, Asadollahi P, Taherikalani M, Emaneini M. Characterization of Klebsiella pneumoniae strains producing extended spectrum beta-lactamases and AMPC type beta-lactamases isolated from hospitalized patients in Kerman, Iran. Roum Arch Microbiol Immunol 2012; 71(2): 81-6.
- Mansouri S, Neyestanaki DK, Shokoohi M, Halimi S, Beigverdi R, Rezagholezadeh F, et al. Characterization of AmpC, CTX-M and MBLs types of ß-lactamases in clinical isolates of Klebsiella pneumoniae and Escherichia coli producing Extended Spectrum ß-lactamases in Kerman, Iran. Jundishapur J Microbiol 2014; 7(2): e8756.
- Song W, Jeong SH, Kim JS, Kim HS, Shin DH, Roh KH, et al. Use of boronic acid disk methods to detect the combined expression of plasmid-mediated AmpC beta-lactamases and extended-spectrum beta-lactamases in clinical isolates of Klebsiella spp., Salmonella spp., and Proteus mirabilis. Diagn Microbiol Infect Dis 2007; 57(3): 315-8.
- Rodriguez-Martinez JM, Poirel L, Nordmann P. Molecular epidemiology and mechanisms of carbapenem resistance in Pseudomonas aeruginosa. Antimicrob Agents Chemother 2009; 53(11): 4783-8.