PHENOTYPIC AND GENOTYPIC CHARACTERISATION OF CARBAPENEM RESISTANT Klebsiella pneumoniae CLINICAL ISOLATES FROM A TERTIARY CARE HOSPITAL

Main Article Content

B. ADITI PRIYADARSHINI
KRISHNAN MAHALAKSHMI

Abstract

Background: The evolution of multidrug resistant Klebsiella pneumoniae is a global threat in treating nosocomial infections. Carbapenem resistant Klebsiella pneumoniae (CRKP) are a group of emerging highly drug-resistant Gram-negative bacilli that produces class A, B and D beta lactamases which help in hydrolyzing carbapenems thereby warranting alternate drug   therapy. This study was aimed to determine the prevalence of CRKP phenotypically and genotypically.

Methods: The identification and antibiotic susceptibility testing of K. pneumoniae isolates were performed by vitek 2 systems version 7.0 as per CLSI guidelines. Phenotypic detection of carbapenemase was performed by combined-disc test using meropenem alone and with both phenyl boronic acid and EDTA. Genotypic detection was performed by multiplex PCR and conventional PCR. The blaKPC, blaNDM, blaOXA-48, blaIMP, blaVIM, blaSPM, blaGIM and blaSIM genes were screened.

Results: The MIC value obtained for carbapenems by vitek 2 showed carbapenem resistance in 19 isolates. All these isolates were positive phenotypically. While, only 11 isolates were positive genotypically showing blaNDM (n=2), blaOXA-48 (n=7) and blaIMP (n=4) gene. All the isolates were susceptible to colistin. Most of the isolates were resistant to cephalosporins and significant resistance to tigecycline was also observed.

Conclusion: The emergence and spread of CRKP is a global threat which hampers the effective control of infections particularly in developing countries. The present study has shown a low prevalence (9.09%) of carbapenem resistance. However the resistance of clinically isolated carbapenem resistant strains can primarily be ascribed to production of carbapenemases there are also other mechanisms like deletion of OMP proteins, production of beta lactamases like ESBL or AmpC contributing significantly. Judicious use of antibiotics and surveillance to detect CRKP strains early will help in reducing the spread of such strains.

Keywords:
Antimicrobial resistance, carbapenemases, carbapenem resistance, multi drug resistance, Klebsiella pneumoniae.

Article Details

How to Cite
PRIYADARSHINI, B. A., & MAHALAKSHMI, K. (2020). PHENOTYPIC AND GENOTYPIC CHARACTERISATION OF CARBAPENEM RESISTANT Klebsiella pneumoniae CLINICAL ISOLATES FROM A TERTIARY CARE HOSPITAL. PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY, 21(7-8), 27-35. Retrieved from http://www.ikprress.org/index.php/PCBMB/article/view/4980
Section
Original Research Article

References

Bratu S, Landman D, Haag R, Recco R, Eramo A, Alam M, et al. Rapid Spread of Carbapenem-Resistant Klebsiella pneumoniae in New York City. Archives of Internal Medicine [Internet]. American Medical Association (AMA). 2005; 165(12):1430.
Available:http://dx.doi.org/10.1001/archinte.165.12.1430

Oteo J, Cuevas O, Lopez-Rodriguez I, Banderas-Florido A, Vindel A, Perez-Vazquez M, et al. Emergence of CTX-M-15-producing Klebsiella pneumoniae of multilocus sequence types 1, 11, 14, 17, 20, 35 and 36 as pathogens and colonizers in newborns and adults. Journal of Antimicrobial Chemotherapy [Internet]. Oxford University Press (OUP); 2009; 64(3):524–8.
Available:http://dx.doi.org/10.1093/jac/dkp211

Gupta A, Ampofo K, Rubenstein D, Saiman L. Extended spectrum β lactamase-producing Klebsiella pneumoniae Infections: A review of the literature. Journal of Perinatology [Internet]. Springer Science and Business Media LLC. 2003; 23(6):439–43.
Available:http://dx.doi.org/10.1038/sj.jp.7210973

Nathisuwan S, Burgess DS, Lewis JS. Extended-spectrum β-lactamases: epidemiology, detection, and treatment. Pharmacotherapy [Internet]. Wiley. 2001; 21(8):920–8.
Available:http://dx.doi.org/10.1592/phco.21.11.920.34529

Meyer KS. Nosocomial outbreak of Klebsiella infection resistant to late-generation cephalosporins. Annals of Internal Medicine [Internet]. American College of Physicians; 1993;119(5):353.
Available:http://dx.doi.org/10.7326/0003-4819-119-5-199309010-00001

Schwaber MJ, Lev B, Israeli A, Solter E, Smollan G, Rubinovitch B, et al. Containment of a country-wide outbreak of carbapenem-resistant Klebsiella pneumoniae in Israeli Hospitals via a Nationally Implemented Intervention. Clinical Infectious Diseases [Internet]. Oxford University Press (OUP). 2011; 52(7):848–55.
Available:http://dx.doi.org/10.1093/cid/cir025

Wang Z, Qin R-R, Huang L, Sun LY. Risk factors for carbapenem-resistant Klebsiella pneumoniae infection and mortality of Klebsiella pneumoniae infection. Chinese Medical Journal [Internet]. Ovid Technologies (Wolters Kluwer Health); 2018;131(1):56–62.
Available:http://dx.doi.org/10.4103/0366-6999.221267

European Centre for Disease Prevention and Control. Risk assessment on the spread of carbapenemase-producing Enterobacteriaceae (CPE) through patient transfer between healthcare facilities, with special emphasis on cross-border transfer. Stockholm: ECDC; 2011.

Cristina ML, Sartini M, Ottria G, Schinca E, Cenderello N, Crisalli MP, Fabbri P, Lo Pinto G, Usiglio D, Spagnolo AM. Epidemiology and biomolecular characteri-zation of carbapenem-resistant Klebsiella pneumoniae in an Italian hospital. J. Prev. Med. Hyg. 2016;57:E149–E156

Poirel L, Walsh TR, Cuvillier V, Nordmann P. Multiplex PCR for detection of acquired carbapenemase genes. Diagnostic Microbiology and Infectious Disease [Internet]. Elsevier BV. 2011;70(1):119– 23.
Available:http://dx.doi.org/10.1016/j.diagmicrobio.2010.12.002

Tsakris A, Poulou A, Pournaras S, Voulgari E, Vrioni G, Themeli- Digalaki K, et al. A simple phenotypic method for the differentiation of metallo-_-lactamases and class A KPC carbapenemases in Enterobacteriaceae clinical isolates. J. Antimicrob. Chemother. 2010;65:1664-1671.

Logan LK, Weinstein RA. The epidemiology of carbapenem-resistant enterobacteriaceae: The impact and evolution of a global menace. The Journal of Infectious Diseases [Internet]. Oxford University Press (OUP). 2017; 215(Suppl_1):S28–S36.
Available:http://dx.doi.org/10.1093/infdis/jiw282

Ito H, Arakawa Y, Ohsuka S, Wacharotayankun R, Kato N, Ohta M. Plasmid mediated dissemination of the metallo-beta-lactamase gene blaIMP among clinically isolated strains of Serratia marcescens. Antimicrob Agents Chemother. 1995;39(4):824-9.

Nordmann P, Naas T, Poirel L. Global spread of carbapenemase-producing Enterobacteriaceae. Emerg Infect Dis. 2011;17:1791–8.

Walsh TR, Toleman MA, Poirel L ,Nordmann P. Metallo-ß-lactamases: The quiet before the storm? Clin Microbiol Rev. 2005;18:306–25.

Vatopoulos A. High rates of metallo-beta-lactamase-producing Klebsiella pneumoniae in Greece–a review of the current evidence: Euro Surveill. 2008;13:8023.

Yong D, Toleman MA, Giske CG, Cho H S, Sundman K, Lee K, et al. Characterization of a new metallo-β-lactamase gene, blaNDM-1, and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India: Antimicrob Agents and Chemother. 2009; 53(12):5046-54.

Poirel L, Héritier C, Tolün V, Nordmann P. Emergence of oxacillinase-mediated resistance to imipenem in Klebsiella pneumonia: Antimicrob Agents and Chemother. 2004;48(1):15-22.

Carrër A, Poirel L, Yilmaz M, Akan O A, Feriha C, Cuzon G, et al. Spread of OXA-48-encoding plasmid in turkey and beyond. Antimicrob Agents and Chemother. 2010;54(3):1369-73.

Castanheira M, Deshpande LM, Mathai D, Bell JM, Jones RN, Mendes RE. Early dissemination of NDM-1- and OXA-181-producing Enterobacteriaceae in Indian hospitals: Report from the SENTRY Antimicrobial Surveillance Program, 2006-2007. Antimicrob Agents Chemother. 2011; 55:1274-8.

Evans BA, Amyes SG. OXA β-lactamases. Clin Microbiol Rev. 2014;27(2):241-63.

Grundmann H, Glasner C, Albiger B, Aanensen DM, Tomlinson CT, Andrasević AT et al. Occurrence of carbapenemase-producing Klebsiella pneumoniae and Escherichia coli in the European survey of carbapenemase-producing Enterobacteriaceae (EuSCAPE): a prospective, multinational study. Lancet Infect Dis. 2017;17(2):153-63.

Amer WH, Khalil HS, Marwa Ahmed Ali Abd EL Wahab. Risk factors, phenotypic and genotypic characterization of carbapenem resistant Enterobacteriaceae in Tanta University Hospitals, Egypt. Int J Infect Control. 2016;12:i2.

Sharma A, Bakthavatchalam YD, Gopi R, Anandan S, Verghese VP, Veeraraghavan B. Mechanisms of Carbapenem Resistance in K.pneumoniae and E. coli from Bloodstream Infections in India. J Infect Dis Ther. 2016;4:293-8.

Padilla E, Llobet E, Doménech-Sánchez A, Martínez-Martínez L, Bengoechea JA, Albertí S. Klebsiella pneumoniae AcrAB efflux pump contributes to antimicrobial resistance and virulence. Antimicrob Agents Chemother. 2010;54(1):177-83.

Ye Y, Xu L, Han Y, Chen Z, Liu C, Ming L. Mechanism for carbapenem resistance of clinical Enterobacteriaceae isolates. Experimental and Therapeutic Medicine [Internet]. Spandidos Publications. 2017;10.
Available:http://dx.doi.org/10.3892/etm.2017.5485

Mohanty S, Mittal G, Gaind R. Identification of carbapenemase mediated resistance among Enterobacteriaceae bloodstream isolates: A molecular study from India. Indian J Med Microbiol. 2017; 35:421-25

Qureshi ZA, Paterson DL, Potoski BA, Kilayko MC , Sandovsky G, Sordillo E, et al. Treatment outcome of bacteremia due to KPC-producing Klebsiella pneumoniae: superiority of combination antimicrobial regimens. Antimicrob Agents Chemother. 2012;56(4):2108–13.

Sun Y, Cai Y, Liu X, Bai N, Liang B, Wang R. The emergence of clinical resistance to tigecycline. Int J Antimicrob Agents. 2013;41:110–6.

Van Duin D, Cober E, Richter SS, Perez F, Cline M, Kaye KS, et al. Tigecycline therapy for carbapenem-resistant Klebsiella pneumoniae (CRKP) bacteriuria leads to tigecycline resistance. Clin Microbiol Infect. 2014;20:O1117–20.

Abouzeed YM, Baucheron S, Cloeckaert A, Ram R. Mutations involved in efflux-mediated multidrug resistance in Salmonella enterica serovar Typhimurium. Antimicrob Agents Chemother. 2008;52:2428–34.

Veleba M, Higgins PG, Gonzalez G, Seifert H, Schneiders T. Characterization of RarA, a novel AraC family multidrug resistance regulator in Klebsiella pneumoniae. Antimicrob Agents Chemother. 2012;56: 4450–8.

Villa L, Feudi C, Fortini D, Garcia-Fernandez A, Carattoli A. Genomics of KPC-producing Klebsiella pneumoniae sequence type 512 clone highlights the role of RamR and ribosomal S10 protein mutations in conferring tigecycline resistance. Antimicrob Agents Chemother. 2013;58:1707–12.

Ahn C, Yoon SS, Yong TS, Jeong SH, Lee K. The resistance mechanism and clonal distribution of tigecycline-nonsusceptible Klebsiella pneumoniae isolates in Korea. Yonsei Med J. 2016;57:641– 6.

Lombardi F, Gaia P, Valaperta R, Cornetta M, Tejada MR, Girolamo LD, et al. Emergence of carbapenem-resistant Klebsiella pneumoniae: Progressive spread and four-year period of observation in a cardiac surgery division. BioMed Res Int. 2015;Article ID 871947:7.