Journal of Medical Bacteriology 2013. 2(3-4):26-31.

Evaluation of Phenotypic Methods for Detection of Klebsiella Pneumoniae Carbapenemase-Producing K. Pneumoniae in Tehran
Leila Azimi, Abdolaziz Rastegar-Lari, Malihe Talebi, Amirmorteza Ebrahimzadeh-Namvar, Somayeh Soleymanzadeh-Moghadam

Abstract


Background: One of the main mechanisms of resistance to carbapenems is potential of Klebsiella pneumoniae to produce K. pneumoniae Carbapenemase (KPC). KPC is an important type of carbapenemase, which can hydrolyze carbapenems and other beta-lactam antibiotics. Modified Hodge Test (MHT) and  use  of  boronic  acid  as  a  KPC  inhibitor  are  two  types  of  phenotypic methods, which are used for detection of carbanemase-producing bacteria. Specificity of these two phenotypic tests for identification of KPC was assessed in this study.
Methods:   Forty-four   K.   pneumoniae   strains   were   isolated   from  wound infections   of   burn   patients.   All   isolates   were   identified   with   specific biochemical tests. Carbapenem-resistant K. pneumoniae isolates were identified by disc diffusion method and analyzed with cut off-points of CLSI 2011 guideline.   For   detection   of   KPC-producing   strains,   carbapenem-resistant isolates were examined with two different phenotypic (i.e. MHT and Boronic acid) methods. Subsequently, strains with positive phenotypic methods were examined by PCR as a molecular method.
Results: Twenty-eight (64%) out of 44 isolates were resistant to carbapenem according  to  CLSI  breakpoints  and  16  (36%)  were  susceptible.  MHT  was positive in all of carbapenem-resistant isolates but none of them have had the synergism effect between meropenem and boronic acid. Also, all isolates were negative for presence of KPC genes on gel electrophoresis. According to results MHT has not enough specificity for detection of KPC.


Keywords


Carbapenemase, Klebsiella pneumoniae, Polymerase Chain, Reaction

Full Text:

PDF

References


Tsakris A, Poulou A, Themeli- Digalaki K et al. Use of Boronic Acid Disc Tests To Detect Extended- Spectrum - Lactamases in Clinical Isolates of KPC Carbapenemase- Possessing Enterobacteriaceae. J Clin Microbiol 2009; 47 (11): 3420-6.

Tsakris A, Kristo I, Poulou A, et al.Evaluation of Boronic Acid Disc Tests for Differentiating KPC-Possessing Klebsiella pneumoniae Isolates in the Clinical Laboratory. J Clin Microbiol 2009; 47 (2): 362-7.

Hirsch EB, Tam VH. Detection and treatment options for Klebsiella pneumoniae carbapenemases (KPCs): an emerging cause of multidrug- resistant infection. J Antimicrob Chemother 2010; 65 (5): 119-25.

Tsakris A, Themeli-Digalaki K, Poulou A, et al. Comparative Evaluation of Combined-Disc Tests Using Different Boronic Acid Compounds for Detection of Klebsiella pneumoniae Carbapenemase Producing Enterobacteriaceae Clinical Isolates. J Clin Microbiol 2011; 49 (8):2804-9.

Rastegar Lari A, Azimi L, Rahbar M, et al. Phenotypic detection of Klebsiella pneumoniae carbapenemase among burns patients: First report from Iran. Burns 2013; 39: 174-6.

Clinical Laboratory Standards Institute.2011. Performance standards for antimicrobial susceptibility testing: nineteenth informational supplement M100-S21. CLSI, Wayne, PA.

Girlich D, Poirel L, Nordmann p- value of the modified Hodge test for detection of emerging carbapenemases in Enterobacteriaceae. J Clin Microbiol 2012; 50 (2): 477-9.

Anderson KF, Lonsway DR, Rasheed JK, et al. Evaluation of methods to identify the Klebsiella pneumoniae carbapenemase in Enterobacteriaceae. J Clin Microbiol 2007; 45 (8): 2723-5.

Tsakris A, Kristo I, Poulou A, et al.The first occurrence of KPC-2- possessing Klebsiella pneumoniae in a Greek hospital and recommendation for detection with boronic acid disc tests. J Antimicrob Chemother 2008;62: 1257-60

Tsakris A, Poulou1 A, Pournaras S, et al. A simple phenotypic method for the differentiation of metallo-b- lactamases and class A KPC carbapenemases in Enterobacteriaceae clinical isolates. J Antimicrob Chemother 2010; 65: 1664-71.

Wolter DJ, Khalaf N, Robledo IE, et al. Surveillance of carbapenem- resistant Pseudomonas aeruginosa isolates from Puerto Rican Medical Center Hospitals: dissemination of KPC and IMP-18 beta-lactamases. Antimicrob Agents Chemother 2009;53 (4): 1660-4.

Brandon Kitchel, J. Kamile Rasheed, Andrea Endimiani, et al. Genetic Factors Associated with Elevated Carbapenem Resistance in KPC- Producing Klebsiella pneumoniae. Antimicrob Agents Chemother0 2010;54 (10): 4201-7.

Thierry Naas, Gaelle Cuzon, Maria- Virginia Villegas, et al. Genetic Structures at the Origin of Acquisition of the β-Lactamase blaKPC Gene. Antimicrob Agents Chemother 2008;52 (4): 1257-63e.

Hesna Yigit, Anne Marie Queenan, Gregory J. Anderson, et al. Novel Carbapenem-Hydrolyzing β-Lactamase, KPC-1, from a Carbapenem-Resistant Strain of Klebsiella pneumoniae. Antimicrob Agents Chemother 2001; 45 (4): 1151-61.

Gomez1 SA, Pasteran1 FG, Faccone D. Clonal dissemination of Klebsiella pneumoniae ST258 harbouring KPC-2 in Argentina. Clin Microbiol Infect 2011; 17 (10): 1520-4.

Nordmann P, Poirel L, Dortet L. Rapid Detection of Carbapenemase producing Enterobacteriaceae. Emerging Infectious Diseases 2012; 18 (9): 1503-7.

Dortet L, Poirel L, Nordmann P.Rapid Detection of Carbapenemase- producing Pseudomonas spp. JCM 2012; 50 (11): 3773-6.

Doi Y, Potoski BA, Adams-Haduch JM, et al. Simple disc-Based Method for Detection of Klebsiella pneumoniae Carbapenemase-Type-Lactamase by Use of a Boronic Acid Compound. J Clin Microbiol 2008; 46 (12): 4083-6.

Anderson KF, Lonsway DR, Rasheed JK, Biddle J, Jensen B, McDougal LK, Carey RB, Thompson A, Stocker S, Limbago B, Patel JB. Evaluation of methods to identify the Klebsiella pneumoniae carbapenemase in Enterobacteriaceae. J Clin Microbiol 2007; 45 (8): 2723-5.

Girlich D, Poirel L, Nordmann P. Value of the modified Hodge test for detection of emerging carbapenemases in Enterobacteriaceae. J Clin Microbiol 2012; 50 (2): 477-9.


Refbacks

  • There are currently no refbacks.


Creative Commons Attribution-NonCommercial 3.0

This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.