Abstract
Background and Objectives: Extended spectrum beta lactamases (ESBLs) are responsible for resistance to third generation Cephalopsorins and Monobactums which form the mainstay of therapy in majority of clinical conditions. This study was performed to generate data on ESBLs and their CTX-M genotypes as prevalent in tertiary care centre at Kottayam, Kerala, India. The prevalence of ESBLs among Gram negative rods in specimens from lower respiratory tract in tertiary care centre at Kottayam was detected using standard methods and their CTX-M genotypes were detected using RT-PCR. The risk factors associated with the emergence of ESBLs and the spread of CTX-M enzymes were discussed.
Objectives: To determine the phenotype prevalence of ESBLs and genotype prevalence of CTX-M among Gram negative rods in specimens from respiratory tract at Government Medical College Hospital, Kottayam using standard methods and RT-PCR respectively.
Materials and Methods: 1216 specimens from lower respiratory tract namely sputum, broncholaveolar lavage and tracheal aspirate were collected during June 2017 to November 2017. Culture yielded 290 isolates of which 40 were multidrug resistant (MDR) strains. Phenotypic testing of ESBLs was done using Clinical Laboratory Standards Institute (CLSI) guidelines- disc diffusion testing. Genotypic testing of CTX-M Beta-lactamase was done using RT-PCR using primers from Gen Bank for Group I- CTX-M1, Group II- TOHO1, Group III- CTX-M825 and Group IV- CTX-M914.
Results: Phenotypic testing showed that out of 40 MDR strains, all 40 tested positive, of which 31/220 (14%) were Klebsiella pneumoniae and 9/70 (12.8%) were E coli. Genotypic testing showed that of the 40 MDR strains, 15% of Klebsiella pneumoniae and 11% of E coli tested positive for Group III CTX-M while all tested negative for Group IV CTX-M. 10-14% of E coli and Klebsiella pneumoniae isolates tested positive for Groups I and II of CTX-M.
Conclusion: Even though CTX-M ESBLs which originated in Kluyvera species were first reported from Japan in 1980s, over the last decades, these genes have dispersed globally among Gram negative rods creating chaos in chemotherapy. This study of lower respiratory tract specimens from tertiary care centre reveals prevalence of ESBLs among multidrug resistant(MDR) strains at 72.7% in phenotype testing and 0-100% in the four gene clusters of CTX-M in genotype testing. Since the clinical microbiology laboratory is the first line of defence in the detection and control of ESBLs and failure to detect ESBLs implies treatment failure, this study stresses the importance of routine ESBL testing of atleast MDR strains and the need for clinicians to practise antibiotic stewardship earnestly.
Keywords: Extended spectrum beta lactamases (ESBLs), CTX-M gene, Multidrug resistant (MDR).
References
- Ambler R. P. (1980). The structure of β-lactamases. Philos.Trans. R. Soc. Lond. B Biol. Sci. 289, 321–33110.1098/ rstb 1980.0049
- Bauernfeind A., Grimm H., Schweighart S. (1990). A new plasmidic cefotaximase in a clinical isolate of Escherichia coli. Infection 18, 294–29810.1007/BF01644637
- Bush K., Jacoby G. A., Medeiros A. A. (1995). A functional classification scheme for β-lactamases and its correlation with molecular structure. Antimicrob. Agents Chemother. 39, 1211–1233
- Canton, R et al. Epidemiology of extended-spectrum β-lactamase-producing Enterobacterisolates in a Spanish hospital during a 12-year period. J. Clin. Microbiol. 40:1237-1243.
- Cantón R., Coque T. M. (2006). The CTX-M β-lactamase pandemic. Curr. Opin. Microbiol. 9, 466–47510.1016/j.mib.2006.08.011
- Canton R, JM Gonzalez-Alba, JC Galan. CTX-M Enzymes: Origin and Diffsuion. Frontiers in Microbiology. 2012; 3:110
- Clinical and Laboratory Standard Institute. Performance standards for antimicrobial susceptibility 22ndinformational supplement CLSI document M100-S22. Vol 32 No.3, 2012.
- Coelho A et al (2010). Characterisation of the CTX-M-15-encoding gene in Klebsiella pneumoniae strains from the Barcelona metropolitan area: plasmid diversity and chromosomal integration. J. Antimicrob. Agents 36, 73–7810.1016/j.ijantimicag.2010.03.005
- Coque T. M., Baquero F., Cantón R. (2008a). Increasing prevalence of ESBL-producing Enterobacteriaceae in Europe. Euro Surveill. 13, 19044.
- Davies J., Davies D. (2010). Origins and evolution of antibiotic resistance. Microbiol. Mol. Biol. Rev. 74, 417–43310.1128/MMBR.00016-10
- Díaz M. A et al Spanish Group for Nosocomial Infections (GEIH) (2010). Diversity of Escherichia colistrains producing extended-spectrum β-lactamases in Spain: second nationwide study. Clin. Microbiol.48, 2840–284510.1128/JCM.02147-09
- Ishii, Y., A. Ohno, H. Taguchi, S. Imajo, M. Ishiguro, and H. Matsuzawa. Cloning and sequence of the gene encoding a cefotaxime-hydrolyzing class A β-lactamase isolated from Escherichia coli. Antimicrob. Agents Chemother. 39:2269-2275.
- Jacoby G. A., Muñoz-Price L. S. (2005). The new β-lactamases. Engl. J. Med.352, 380–39110.1056/NEJMra041359
- Kronenberg A, Hilty M, Endimiani A, Mühlemann K. Temporal trends of extended-spectrum cephalosporin-resistant Escherichia coliand Klebsiella pneumoniae isolates in in- and outpatients in Switzerland, 2004 to 2011. Euro Surveill. 2013;18(21):pii=20484.
- Mathur P, Kapil A, Das B, Dhawan B. Prevalence of ESBL producing Gram negative bacteria in a tertiary care hospital Indian J Med Res. 2002;115:153–57.
- Mendelson G, Hait V, Ben-Israel J, Gronich D, Granot E, Raz R. Prevalence and risk factors of extended-spectrum beta-lactamase-producing Escherichia coliand Klebsiella pneumoniae in an Israeli long-term care facility. European Journal of Clinical Microbiology & Infectious Diseases. 2004;24(1):17–22.
- National Committee for Clinical Laboratory Standards. Performance standards for antimicrobial and susceptibility testing. Twelfth informational supplement. M100-S12. National Committee for Clinical Laboratory Standards, Wayne, Pa.
- Pitout, J. D., N. D. Hanson, D. L. Church, and K. B. Laupland. Population-based laboratory surveillance for Escherichia coli-producing extended-spectrum beta-lactamases: importance of community isolates with blaCTX-M genes. Clin. Infect. Dis. 38:1736-1741.
- Pitout, J. D., A Hossain, N. D. Hanson. 2004. Phenotypic and Molecular Detection of CTX-M-Beta lactamases produced by E coli and Klebsiella spp. J Clin Microbiol 2004 Dec 42(12): 5715-5721.
- Rodríguez M. M et al (2004). Chromosome-encoded CTX-M-3 from Kluyvera ascorbata: a possible origin of plasmid-borne CTX-M-1-derived cefotaximases. Agents Chemother.48, 4895–489710.1128/AAC.48.12.4895-4897.2004
- Rodríguez M M et al(2010). Novel chromosome-encoded CTX-M-78 β-lactamase from a Kluyvera Georgiana clinical isolate as a putative origin of CTX-M-25 subgroup. Agents Chemother.54, 3070–307110.1128/AAC.01615-09
- Rodriguez-Villalobos H et al. (2011).Trends in production of extended-spectrum β-lactamases among Enterobacteriaceae of clinical interest: results of a nationwide survey in Belgian hospitals. Antimicrob. Chemother.66, 37–4710.1093/jac/dkq388
- Sharma M. Prevalence and antibiogram of Extended Spectrum β-Lactamase (ESBL) producing Gram negative bacilli and further molecular characterization of ESBL producing Escherichia coliand Klebsiell a Journal of Clinical and Diagnostic Research. 2013;7(10):2173–77.
- Zhanel GG, Decorby M, Laing N, Weshnoweski B, Vashisht R, Tailor F, et al. Antimicrobial-Resistant pathogens in intensive care units in canada: results of the canadian national intensive care unit (CAN-ICU) Study, 2005-2006. Antimicrobial Agents and Chemotherapy. 2008;52 (4):1430–37.
Corresponding Author
Sheeba K Thomas
Assistant Professor, Department of Microbiology,
Government Medical College, Kottayam, Kerala, India