Phenotypic and genotypic characterization of carbapenem resistance mechanisms in Klebsiella pneumoniae from blood culture specimens: A study from North India

Varsha Gupta; Ritu Garg; Karthikeyan Kumaraswamy; Priya Datta; Gursimran Kaur Mohi; Jagdish Chander

doi:10.4103/JLP.JLP_155_16

Journal of Laboratory Physicians, Table of Contents

CC BY-NC-ND 4.0 · J Lab Physicians 2018; 10(02): 125-129
DOI: 10.4103/JLP.JLP_155_16

Original Article

Phenotypic and genotypic characterization of carbapenem resistance mechanisms in Klebsiella pneumoniae from blood culture specimens: A study from North India

Varsha Gupta

Department of Microbiology, Government Medical College Hospital, Chandigarh, India

,

Ritu Garg

Department of Microbiology, Government Medical College Hospital, Chandigarh, India

,

Karthikeyan Kumaraswamy

Department of ARM Laboratory, Erode, Tamil Nadu, India

,

Priya Datta

Department of Microbiology, Government Medical College Hospital, Chandigarh, India

,

Gursimran Kaur Mohi

Department of Microbiology, Government Medical College Hospital, Chandigarh, India

,

Jagdish Chander

Department of Microbiology, Government Medical College Hospital, Chandigarh, India

› Author Affiliations

Abstract

PDF Download

ABSTRACT

BACKGROUND: Emergence of carbapenem resistance among Enterobacteriaceae in different geographical regions is of great concern as these bacteria are easily transmissible among patients. Carbapenem-resistance in Enterobacteriaceae is due to production of carbapenemases of various classes and hyper production of the ESBLs (Extended spectrum beta lactamases) and Amp C beta lactamases with reduced cell wall permeability mechanisms. Phenotypic detection and differentiation is important for proper infection control and appropriate patient management. This study was done to know the prescence of various beta lactamases and carbapenemases with other mechanisms of resistance in Klebsiella pneumoniae isolates.

MATERIALS AND METHODS: 50 non-duplicate carbapenem resistant isolates of Klebsiella pneumoniae from blood culture specimens were included and various mechanisms of resistance were studied based on phenotypic and genotypic methods.

RESULTS: Out of 50 isolates, 39 (78%) of K.pneumoniae isolates were Extended Spectrum Beta Lactamase (ESBL) producers based on CLSI guidelines. All 50 showed positive Modified Hodge Test (MHT ) and 32 showed Metallo Beta Lactamase (MBL) by Combined Disc Test (CDT). Four isolates showed AmpC production with porin loss . None of the isolates showed Class A KPC production by CDT . In our study all the 10 isolates evaluated by genotypic technique produced CTX-M group 1 enzyme by multiplex PCR. Seven out of 10 strains which showed positive MBL results were positive for NDM .

CONCLUSIONS: Carbapenems are often considered last resort antibiotics in the treatment of infections due to multidrug-resistant organisms. It is therefore mandatory to maintain the clinical efficacy of carbapenems by early detection of various enzymes . For routine clinical laboratories both phenotypic and genotypic tests need to be followed to detect various mechanisms of carbapenem resistance and this is of epidemiological relevance also.

Key words

Amp C - carbepenem - Enterobacteriaceae

PDF (448 kb)

References

REFERENCES
1 Gupta N, Limbago BM, Patel JB, Kallen AJ. Carbapenem-resistant Enterobacteriaceae: Epidemiology and prevention. Clin Infect Dis 2011;53:60-7.
2 World Health Organization. Antimicrobial Resistance: Global Report on Surveillance; 2014. Available from: http://www.who. int/drugresistance/documents/surveillancereport/en/. [Last accessed on 2015 Jun 15].
3 Bakthavatchalam YD, Anandan S, Veeraraghavan B. Laboratory detection and clinical implication of oxacillinase-48 like carbapenemase: The hidden threat. J Glob Infect Dis 2016;8:41-50.
4 Kazi M, Drego L, Nikam C, Ajbani K, Soman R, Shetty A, et al. Molecular characterization of carbapenem-resistant Enterobacteriaceae at a tertiary care laboratory in Mumbai. Eur J Clin Microbiol Infect Dis 2015;34:467-72.
5 Chen L, Mathema B, Chavda KD, DeLeo FR, Bonomo RA, Kreiswirth BN. Carbapenemase-producing Klebsiella pneumoniae: Molecular and genetic decoding. Trends Microbiol 2014;22:686-96.
6 Kumarasamy K, Kalyanasundaram A. Emergence of Klebsiella pneumoniae isolate co-producing NDM-1 with KPC-2 from India. J Antimicrob Chemother 2012;67:243-4.
7 Queenan AM, Bush K. Carbapenemases: The versatile beta-lactamases. Clin Microbiol Rev 2007;20:440-58.
8 Collee JG, Miles RS, Watt B. Tests for the identification of bacteria. In: Collee JG, Fraser AG, Marmion BP, Simmons A, editors. Mackie & MacCartney Practical Medical Microbiology. 14^th ed. London: Church ill Living Stone; 1996. p. 151-79.
9 Wayne PA. Clinical and Laboratory Standards Institute: Performance Standards for Antimicrobial Susceptibility Testing; 25^th Informational Supplement. CLSI Document M100-S25; 2015.
10 Giske CG, Gezelius L, Samuelsen Ø, Warner M, Sundsfjord A, Woodford N. A sensitive and specific phenotypic assay for detection of metallo-ß-lactamases and KPC in Klebsiella pneumoniae with the use of meropenem disks supplemented with aminophenylboronic acid, dipicolinic acid and cloxacillin. Clin Microbiol Infect 2011;17:552-6.
11 Manchanda V, Singh NP. Occurrence and detection of AmpC beta-lactamases among Gram-negative clinical isolates using a modified three-dimensional test at Guru Tegh Bahadur Hospital, Delhi, India. J Antimicrob Chemother 2003;51:415-8.
12 Lee W, Jung B, Hong SG, Song W, Jeong SH, Lee K, et al. Comparison of 3 phenotypic-detection methods for identifying plasmid-mediated AmpC beta-lactamase-producing Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis strains. Korean J Lab Med 2009;29:448-54.
13 Woodford N, Fagan EJ, Ellington MJ. Multiplex PCR for rapid detection of genes encoding CTX-M extended-spectrum (beta)-lactamases. J Antimicrob Chemother 2006;57:154-5.
14 Pérez-Pérez FJ, Hanson ND. Detection of plasmid-mediated AmpC beta-lactamase genes in clinical isolates by using multiplex PCR. J Clin Microbiol 2002;40:2153-62.
15 Gupta V, Kumarasamy K, Gulati N, Garg R, Krishnan P, Chander J. AmpC ß-lactamases in nosocomial isolates of Klebsiella pneumoniae from India. Indian J Med Res 2012;136:237-41.
16 Tzouvelekis LS, Markogiannakis A, Piperaki E, Souli M, Daikos GL. Treating infections caused by carbapenemase-producing Enterobacteriaceae. Clin Microbiol Infect 2014;20:862-72.
17 Asthana S, Mathur P, Tak V. Detection of carbapenemase production in gram-negative bacteria. J Lab Physicians 2014;6:69-75.
18 Datta P, Gupta V, Garg S, Chander J. Phenotypic method for differentiation of carbapenemases in Enterobacteriaceae: Study from north India. Indian J Pathol Microbiol 2012;55:357-60.
19 Chauhan K, Pandey A, Asthana AK, Madan M. Evaluation of phenotypic tests for detection of Klebsiella pneumoniae carbapenemase and metallo-beta-lactamase in clinical isolates of Escherichia coliand Klebsiella species. Indian J Pathol Microbiol 2015;58:31-5.
20 Mohan B, Hallur V, Singh G, Sandhu HK, Appannanavar SB, Taneja N. Occurrence of blaNDM-1 and absence of blaKPC genes encoding carbapenem resistance in uropathogens from a tertiary care centre from North India. Indian J Med Res 2015;142:336-43.
21 Maurer FP, Castelberg C, Quiblier C, Bloemberg GV, Hombach M. Evaluation of carbapenemase screening and confirmation tests with Enterobacteriaceae and development of a practical diagnostic algorithm. J Clin Microbiol 2015;53:95-104.
22 van Dijk K, Voets GM, Scharringa J, Voskuil S, Fluit AC, Rottier WC, et al. Adisc diffusion assay for detection of class A, B and OXA-48 carbapenemases in Enterobacteriaceae using phenyl boronic acid, dipicolinic acid and temocillin. Clin Microbiol Infect 2014;20:345-9.
23 Wang P, Chen S, Guo Y, Xiong Z, Hu F, Zhu D, et al. Occurrence of false positive results for the detection of carbapenemases in carbapenemase-negative Escherichia coliand Klebsiella pneumoniae isolates. PLoS One 2011;6:e26356.
24 Woerther PL, Burdet C, Chachaty E, Andremont A. Trends in human fecal carriage of extended-spectrum ß-lactamases in the community: Toward the globalization of CTX-M. Clin Microbiol Rev 2013;26:744-58.
25 Kaur M, Aggarwal A. Occurrence of the CTX-M, SHV and the TEM genes among the extended spectrum ß-lactamase producing isolates of Enterobacteriaceae in a tertiary care hospital of North India. J Clin Diagn Res 2013;7:642-5.
26 Batra P, Dwivedi M, Sherwal BL, Dutta R, Gupta S. NDM-1 Infection and colonisation in critically ill patients from Delhi: A glimpse of the community scenario. Indian J Med Microbiol 2016;34:120-1.