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Detection of AmpC β-Lactamase in Gram Negative Bacteria with their Predisposing Factors and Antimicrobial Susceptibility Pattern


Affiliations
1 Dept of Microbiology, MGM Medical College & Hospital, Navi Mumbai, India
2 Dept of Microbiology, Topiwala National Medical College & BYL Nair Hospital, Mumbai Central, Mumbai, India
3 Dept of Medicine, MGM Medical College & Hospital, Navi Mumbai, India
     

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Introduction: The predominant mechanism for resistance to β-lactam antibiotics in gram-negative bacteria is the synthesis of β-lactamase enzyme. Production of AmpC β-lactamase enzyme is a major mechanism of resistance in many Gram-negative bacteria. There is not much Indian data on the prevalence of AmpC β-lactamases in Gram-negative bacteria. AmpC β-lactamase was reported in many parts of the world like USA (85.5%), China (16.9%), Singapore (49.8%), Korea (29.6%), etc. and also some parts of India like New Delhi (20.7%), Chennai (37.5%), Aligarh (20%), Kolkata (17.3%), Bangalore (42.85%), Varanasi (22%), Pondicherry (47%), etc. However, no documented report is available in and around Mumbai.

Materials and Methods: This is a study conducted in Lokmanya Tilak Municipal Medical College and General Hospital, Sion, Mumbai. The same is attributed to the population living in Sion and nearby localities. Two hundred non-duplicate isolates of Gram negative bacteria recovered from different clinical specimens like wound swab, urine, Foley's catheter tip, blood, sputum from patients admitted in the hospital were tested for AmpC β-lactamase production by Modified Double Disk approximation Method, AmpC Disk Test, Modified Three Dimensional Test.

Results: Overall prevalence of AmpC β-lactamase by MDDM was 62%, AmpC β-lactamase by AmpC disk test was 40%, AmpC β-lactamase was 40%. All the isolates tested were resistant to cefoxitin. Multi Drug Resistance (MDRs) was seen in 15% (30/200) isolates. Maximum AmpC positive isolates were E. coli (36.25%), followed by K.pneumoniae (26.25%). No imipenem resistance was seen in AmpC producers. The patients with bacteria producing AmpC β-lactamase had multiple risk factors like Duration of hospital stay > 8days, urinary catheterization, central line insertion, previous antibiotic use, mechanical ventilation, etc. Patients with AmpC positive isolates who survived and responded to imipenem therapy was 93.75% and mortality rate was 6.25%.

Conclusion: The present study emphasizes the high prevalence (40%) of AmpC β-lactamases in Gram-negative bacteria. Early detection of these AmpC β-lactamase producing isolates in a routine laboratory would help avoid treatment failure. Inability to detect AmpC β-lactamase contributes to their uncontrolled spread and therapeutic failure. Hence their appearances in hospital setting should be identified promptly, so that appropriate antibiotic usage and containment measures can be implemented.

Antibiotic exposure may probably be the main risk factor for acquisition of AmpC β-lactamases. So, avoiding unnecessary antibiotic therapy may be the most important factor to prevent infections in patients with AmpC positive isolates. Therefore, strict antibiotic policies and measures to limit the indiscriminate use of cephalosporins and carbapenems in hospital environments should be undertaken to minimize the emergence and spread of AmpC β-lactamase producing bacteria.


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  • Jacoby GA. AmpC β-Lactamases. Clinical Microbio-logy Reviews. 2009; 22(1): 161-82
  • Hall BG, Barlow M. Revised Ambler classification of β-lactamases. Journal of Antimicrobial Chemotherapy. 2005; 55(6): 1050-1
  • Bush K, Jacoby GA Medeiros AA. A functional classification scheme for β-lactamases and its correlation with molecular structure. Antimicrob Agents Chemother. 1995; 39(6): 1211-33.
  • Bush K, Macalintal C, Rasmussen BA, Lee VJ, Yang Y. Kinetic interactions of tazobactam with β-lactamases from all major structural classes. Antimicrob Agents Chemother. 1993; 37(4): 851-8.
  • Kazmierczak A, Cordin X, Duez JM, Siebor E, Pechinot A, Sirot J. Differences between clavulanic acid and sulbactam in induction and inhibition of cephalosporinases in enterobacteria. J Int Med Res. 1990; 18(Suppl.4): 67D-77D.
  • Monnaie D, Frere JM. Interaction of clavulanate with class C β-lactamases. FEBS Lett. 1993; 334(3): 269-71.
  • Philippon A, Arlet G, Jacoby GA. Plasmiddetermined AmpC-type β-lactamases. Antimicrob Agents Chemother. 2002; 46(1): 1-11.
  • Walther-Rasmussen J, Høiby N. Plasmid-borne AmpC β-lactamases. Can J Microbiol. 2002; 48(6): 479-93.
  • Dunne WM Jr, Hardin DJ. Use of Several Inducer and Substrate Antibiotic Combinations in a Disk Approximation Assay Format To Screen for AmpC Induction in Patient Isolates of Pseudomonas aeruginosa, Enterobacter spp, Citrobacter spp and Serratia spp. J Clin Microbiol. 2005; 43(12): 5945-9.
  • Li Y, Li Q, Du Y, Jiang X, Tang J, Wang J, et al. Prevalence of Plasmid-Mediated AmpC β-Lactamases in a Chinese University Hospital from 2003 to 2005: First Report of CMY-2-Type AmpC β-Lactamase Resistance in China. J Clin Microbiol. 2008; 46(4): 1317-21
  • Tan TY, Ng LSY, He J, Koh TH, Hsu LY. Evaluation of Screening Methods To Detect Plasmid-Mediated AmpC in Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis. Antimicrob Agents Chemother. 2009; 53(1): 146-9
  • Kim J, Lim Yu-Mi. Prevalence of Derepressed AmpC Mutants and Extended-Spectrum β-Lactamase Producers among Clinical Isolates of Citrobacter freundii, Enterobacter spp, and Serratia marcescens in Korea: Dissemination of CTX-M-3, TEM-52 and SHV-12. J Clin Microbiol. 2005; 43(5): 2452-5
  • Manchanda V, Singh NP. Occurrence and detection of AmpC β-lactamases among Gram-negative clinical isolates using a modified three-dimentional test at Guru Tegh Bahadur Hospital, Delhi, India. Journal of Antimicrobial Chemotherapy. 2003; 51(2): 415-8
  • Subha A, Devi VR, Ananthan S. AmpC β-lactamase producing multidrug resistant strains of Klebsiella spp. and Escherichia coli isolated from children under five in Chennai. Indian J Med Res. 2003;117:138
  • Shahid M, Malik A, Agrawal M, Singhal S. Phenotypic detection of extended spectrum and AmpC β-lactamases by a new spot inoculation method and modified three dimensional extract test: Comparison with the conventional three-dimensional extract test. J Antimicrob Chemother. 2004; 54:684-7
  • Arora S, Bal M. AmpC β-lactamases producing bacterial isolates from Kolkata hospital. Indian J Med Res. 2005; 122: 224-33
  • Sinha M. Srinivasa H. Mechanisms of resistance to carbapenems in meropenem-resistant Acinetobacter isolates from clinical samples. Indian J Med Microbiol. 2007; 25(2): 121-5
  • Bhattacharjee A, Anupurba S, Gaur A, Sen MR. Prevalence of Inducible AmpC β-lactamaseProducing Pseudomonas aeruginosa in a Tertiary Care Hospital in Northern India. Indian J Med Microbiol. 2008; 26(1): 89-98
  • Mohamudha PR, Harish BN, Parija SC. AmpC beta lactamases among Gram negative clinical isolates from a tertiary hospital, South India. Braz J Microbiol. 2010; 41(3): Sao Paulo
  • Singhal S, Mathur T, Khan S, Upadhyay DJ, Chugh S, Gaind R, et al. Evaluation of Methods for AmpC Beta- Lactamase in Gram Negative Clinical Isolates from Tertiary Care Hospitals. Indian J Med Microbiol. 2005; 23(2): 120-4
  • Black JA, Moland ES, Thomson KS. AmpC Disk Test for Detection of Plasmid-Mediated AmpC β -Lactamases in Enterobacteriaceae Lacking Chromosomal AmpC β -Lactamases. J Clin Microbiol. 2005; 43(7): 3110–3.
  • Performance Standards for Antimicrobial Susceptibility Testing; Twentieth Informational Supplement. Clinical Laboratory Standards Institute. 2010 M100 - S20; 30(1): pp 40-55
  • Jaurin B, Grundström T. ampC cephalosporinase of Escherichia coli K-12 has a different evolutionary origin from that of β-lactamases of the penicillinase type. Proc Natl Acad Sci USA. 1981; 78(8): 4897-901.
  • Khan MKR, Thukral SS, Gaind R. Evaluation of a modified double-disc synergy test for detection of extended spectrum β-lactamases in AmpC β-lactamase-producing Proteus mirabilis Indian J Med Microbiol. 2008; 26(1): 58-61
  • Coudron PE, Hanson ND, Climo MW. Occurrence of extended-spectrum and AmpC beta-lactamases in bloodstream isolates of K. pneumoniae: isolates harbour plasmid-mediated FOX-5 and ACT-1 AmpC beta-lactamases. J Clin Microbiol. 2003; 41(2): 772-7
  • Javier Perez- Perez F, Hanson ND. Detection of Plasmid- Mediated AmpC β- Lactamase Genes in Clinical Isolates by Using Multiplex PCR. J Clin Microbiol. 2002; 40(6): 2153-62
  • Upadhyay S, Sen MR, Bhattacharjee A. Presence of different β-lactamase classes among clinical isolates of Pseudomonas aeruginosa expressing AmpC beta-lactamase enzyme. J Infect Dev Ctries. 2010; 4(4): 239-42
  • Basak S, Khodke M, Bose S, Mallick SK. Inducible AmpC beta-lactamase producing Pseudomonas aeruginosa isolated in a rural hospital of Central India. Indian J Med Res. 2009; 3(6): 1921-7
  • Sinha P, Sharma R, Rishi S, Sharma R, Sood S, Pathak D. Prevalence of extended spectrum beta lactamase and AmpC beta lactamase producers among Escherichia coli isolates in a tertiary care hospital in Jaipur. Indian J Pathol Microbiol. 2008; 51(3): 367-9
  • Pai H, Kang CI, Byeon JH, Lee KD, Park WB, Kim HB, Kim EC, Oh MD, Choe KW. Epidemiology and clinical features of bloodstream infections caused by AmpC-type-β-lactamase-producing Klebsiella pneumoniae. Antimicrob Agents Chemother. 2004; 48(10): 3720-8.
  • Taneja N, Rao P, Arora J, Dogra A. Occurrence of ESBL and AmpC beta-lactamases and susceptibility to newer antimicrobial agents in complicated UTI. Indian J. Med. Res.2008;
  • Yang K, Guglielmo BJ. Diagnosis and treatment of Expended-Spectrum and AmpC β-lactamase producing organisms. Ann Pharmacother. 2007; 41: 1427-35
  • Alvarez M, Tran JH, Chow N, Jacoby GA. Epidemiology of conjugative plasmid-mediated AmpC β-lactamases in the United States. Antimicrobial Agents and Chemother. 2004; 48(2): 533-7
  • Linares L, Cervera C, Cofan F, Lizaso D, Marco F, Ricart MJ, et al. Risk factors for infections with extended-spectrum and AmpC beta-lactamaseproducing gram-negative rods in renal transplantation. Am J Transplant. 2008; 8(5) : 1000-5
  • Thomson KS. Controversies about extended spectrum and AmpC β- lactamases. Emerging Infectious Diseases. 2001; 7(2):
  • Roh KH, Uh Y, Kim JS, Kim HS, Shin DH, Song W. First outbreak of Multidrug-Resistant Klebsiella pneumoniae producing both SHV-12 Type extendedspectrum β-lactamase and DHA-1 Type AmpC β-lactamase at a Korean hospital. Yonsei Med J. 2008; 49(1): 53-7

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  • Detection of AmpC β-Lactamase in Gram Negative Bacteria with their Predisposing Factors and Antimicrobial Susceptibility Pattern

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Authors

Deepashri Naik
Dept of Microbiology, MGM Medical College & Hospital, Navi Mumbai, India
Anuradha De
Dept of Microbiology, Topiwala National Medical College & BYL Nair Hospital, Mumbai Central, Mumbai, India
Bhaimangesh Naik
Dept of Medicine, MGM Medical College & Hospital, Navi Mumbai, India

Abstract


Introduction: The predominant mechanism for resistance to β-lactam antibiotics in gram-negative bacteria is the synthesis of β-lactamase enzyme. Production of AmpC β-lactamase enzyme is a major mechanism of resistance in many Gram-negative bacteria. There is not much Indian data on the prevalence of AmpC β-lactamases in Gram-negative bacteria. AmpC β-lactamase was reported in many parts of the world like USA (85.5%), China (16.9%), Singapore (49.8%), Korea (29.6%), etc. and also some parts of India like New Delhi (20.7%), Chennai (37.5%), Aligarh (20%), Kolkata (17.3%), Bangalore (42.85%), Varanasi (22%), Pondicherry (47%), etc. However, no documented report is available in and around Mumbai.

Materials and Methods: This is a study conducted in Lokmanya Tilak Municipal Medical College and General Hospital, Sion, Mumbai. The same is attributed to the population living in Sion and nearby localities. Two hundred non-duplicate isolates of Gram negative bacteria recovered from different clinical specimens like wound swab, urine, Foley's catheter tip, blood, sputum from patients admitted in the hospital were tested for AmpC β-lactamase production by Modified Double Disk approximation Method, AmpC Disk Test, Modified Three Dimensional Test.

Results: Overall prevalence of AmpC β-lactamase by MDDM was 62%, AmpC β-lactamase by AmpC disk test was 40%, AmpC β-lactamase was 40%. All the isolates tested were resistant to cefoxitin. Multi Drug Resistance (MDRs) was seen in 15% (30/200) isolates. Maximum AmpC positive isolates were E. coli (36.25%), followed by K.pneumoniae (26.25%). No imipenem resistance was seen in AmpC producers. The patients with bacteria producing AmpC β-lactamase had multiple risk factors like Duration of hospital stay > 8days, urinary catheterization, central line insertion, previous antibiotic use, mechanical ventilation, etc. Patients with AmpC positive isolates who survived and responded to imipenem therapy was 93.75% and mortality rate was 6.25%.

Conclusion: The present study emphasizes the high prevalence (40%) of AmpC β-lactamases in Gram-negative bacteria. Early detection of these AmpC β-lactamase producing isolates in a routine laboratory would help avoid treatment failure. Inability to detect AmpC β-lactamase contributes to their uncontrolled spread and therapeutic failure. Hence their appearances in hospital setting should be identified promptly, so that appropriate antibiotic usage and containment measures can be implemented.

Antibiotic exposure may probably be the main risk factor for acquisition of AmpC β-lactamases. So, avoiding unnecessary antibiotic therapy may be the most important factor to prevent infections in patients with AmpC positive isolates. Therefore, strict antibiotic policies and measures to limit the indiscriminate use of cephalosporins and carbapenems in hospital environments should be undertaken to minimize the emergence and spread of AmpC β-lactamase producing bacteria.


References