Incidence of metallo-beta-lactamase producing clinical isolates of Escherichia coli and Klebsiella pneumoniae in central Nepal

BMC Research Notes

Khilasa Pokharel¹  Surya Narayan Mahaseth¹  Brajesh Kumar Jha¹  Rajkumari Sanjana¹  Arijit Bora¹ 

[1] Department of Microbiology, College of Medical Sciences, P.O. Box 23, Bharatpur, Nepal

关键词: Pandrug-resistant;    Multidrug-resistant;    Carbapenemase;    Metallo-beta-lactamase;    Klebsiella pneumoniae;    Escherichia coli;   

Others  :  1130263
DOI  :  10.1186/1756-0500-7-557

 received in 2014-04-12, accepted in 2014-08-12,  发布年份 2014

【 摘 要 】

Background

The increasing and rapid spread of metallo-beta-lactamase (MBL) producing Enterobacteriaceae, particularly Escherichia coli and Klebsiella pneumoniae represents an emerging public health threat. However, limited data is available on MBL production in clinical isolates of E. coli and K. pneumoniae from Nepal. We have therefore undertaken this study to ascertain the incidence of MBL production in clinical isolates of E. coli and K. pneumoniae at a tertiary care teaching hospital in central Nepal.

Methods

A total of 401 consecutive, non-duplicate isolates of E. coli (n = 216) and K. pneumoniae (n = 185) were recovered from various clinical samples between July and December, 2012. These isolates were screened for the detection of carbapenemase production on the basis of their reduced susceptibility to meropenem or ertapenem by the disc diffusion method. The screened isolates were further phenotypically studied for carbapenemase production by modified Hodge test (MHT). MBL production was detected by performing combined disc test by using imipenem discs with and without ethylenediaminetetraacetic acid (EDTA), which chelates zinc required for MBL activity.

Results

Out of 216 E. coli isolates, a total of 41 isolates (18.98%) and out of 185 K. pneumoniae isolates, a total of 39 isolates (21.08%) were suspected to be carbapenemase- producers on the basis of their reduced susceptibility to meropenem or ertapenem. Interestingly, all the initially suspected isolates of E. coli and K. pneumoniae for carbapenemase production were found to be positive in both MHT and combined disc test. However, few weakly positive reactions were observed in MHT. All the MBL producing isolates were multidrug-resistant (MDR). In addition, 75.60% E. coli and 71.79% of K. pneumoniae isolates producing MBL were found to be “pandrug- resistant”.

Conclusions

Our findings showed MBL production in a considerable number of E. coli and K. pneumoniae isolates with MDR and pandrug-resistant phenotypes. Combined disc method can provide a sensible choice for phenotypic detection of MBL production in clinical microbiology laboratories as detection of MBL in bacterial isolates is indispensable for establishing the effective antibiotic policies and infection control strategies in the hospital setting.

【 授权许可】

   
2014 Bora et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Rasheed JK, Kitchel B, Zhu W, Anderson KF, Clark NC, Ferraro MJ, Savard P, Humphries RM, Kallen AJ, Limbago BM: New Delhi metallo-β-lactamase-producing Enterobacteriaceae, United States. Emerg Infect Dis 2013, 19:870-878.
• [2]Bebrone C: Metallo-beta-lactamases (classification, activity, genetic organization, structure, zinc coordination) and their superfamily. Biochem Pharmacol 2007, 74:1686-1701.
• [3]Ambler RP: The structure of beta-lactamases. Philos Trans R Soc Lond B Biol Sci 1980, 289:321-331.
• [4]Bush K, Jacoby GA: Updated functional classification of β-lactamases. Antimicrob Agents Chemother 2010, 54:969-976.
• [5]Patel G, Bonomo RA: “Stormy waters ahead”: global emergence of carbapenemases. Front Microbiol 2013, 4:48.
• [6]Cornaglia G, Giamarellou H, Rossolini GM: Metallo-β-lactamases: a last frontier for β-lactams. Lancet Infect Dis 2011, 11:381-393.
• [7]Walsh TR: Emerging carbapenemases: a global perspective. Int J Antimicrob Agents 2010, 36(Suppl 3):S8-S14.
• [8]Collee JG, Miles RS, Watt B: Tests for Identification of Bacteria. In Mackie and McCartney Practical Medical Microbiology. 14th edition. Edited by Collee JG, Fraser AG, Marmion BP, Simmons A. New York: Churchill Livingstone; 1996:131-149.
• [9]Clinical Laboratory Standards Institute (CLSI): Performance Standards for Antimicrobial Susceptibility Testing; Twenty First Informational Supplement. Wayne PA: M 100-S21, CLSI; 2011.
• [10]Galani I, Kontopidou F, Souli M, Rekatsina PD, Koratzanis E, Deliolanis J, Giamarellou H: Colistin susceptibility testing by Etest and disc diffusion methods. Int J Antimicrob Agents 2008, 31:434-43 9.
• [11]Franklin C, Liolios L, Peleg AY: Phenotypic detection of carbapenem susceptible metallo-beta-lactamase-producing gram-negative bacilli in the clinical laboratory. J Clin Microbiol 2006, 44:3139-3144.
• [12]Mishra SK, Acharya J, Kattel HP, Koirala J, Rijal BP, Pokhrel BM: Metallo-beta-lactamase producing gram-negative bacterial isolates. J Nepal Health Res Counc 2012, 10:208-213.
• [13]Khanal S, Joshi DR, Bhatta DR, Devkota U, Pokhrel BM: β-lactamase-producing multidrug-resistant bacterial pathogens from tracheal aspirates of intensive care unit patients at national institute of neurological and allied sciences. Nepal ISRN Microbiol 2013, 2013:847569.
• [14]Yong D, Choi YS, Roh KH, Kim CK, Park YH, Yum JH, Lee K, Chong Y: Increasing prevalence and diversity of metallo-beta-lactamases in Pseudomonas spp., Acinetobacter spp., and Enterobacteriaceae from Korea. Antimicrob Agents Chemother 2006, 50:1884-1886.
• [15]Kumar S, Bandyopadhyay M, Mondal S, Pal N, Ghosh T, Bandyopadhyay M, Banerjee P: Tigecycline activity against metallo-β-lactamase-producing bacteria. Avicenna J Med 2013, 3:92-96.
• [16]Datta S, Wattal C, Goel N, Oberoi JK, Raveendran R, Prasad KJ: A ten year analysis of multi-drug resistant blood stream infections caused by Escherichia coli & Klebsiella pneumoniae in a tertiary care hospital. Indian J Med Res 2012, 135:907-912.
• [17]Brusselaers N, Vogelaers D, Blot S: The rising problem of antimicrobial resistance in the intensive care unit. Ann Intensive Care 2011, 1:47.
• [18]Ramana KV, Rao R, Sharada CV, Kareem M, Reddy LR, Ratna Mani M: Modified Hodge test: a useful and the low-cost phenotypic method for detection of carbapenemase producers in Enterobacteriaceae members. J Nat Sci Biol Med 2013, 4:346-348.
• [19]Perry JD, Naqvi SH, Mirza IA, Alizai SA, Hussain A, Ghirardi S, Orenga S, Wilkinson K, Woodford N, Zhang J, Livermore DM, Abbasi SA, Raza MW: Prevalence of the faecal carriage of Enterobacteriaceae with NDM-1 carbapenemase at military hospitals in Pakistan, and the evaluation of two chromogenic media. J Antimicrob Chemother 2011, 66:2288-2294.
• [20]Bora A, Ahmed GU, Hazarika NK, Prasad KN, Shukla SK, Randhawa V, Sarma JB: Incidence of bla NDM-1 gene in Escherichia coli isolates at a tertiary care referral hospital in Northeast India. Indian J Med Microbiol 2013, 31:250-256.
• [21]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, 55:1274-1278.
• [22]Deshmukh DG, Damle AS, Bajaj JK, Bhakre JB, Patwardhan NS: Metallo-β-lactamase-producing clinical isolates from patients of a tertiary care hospital. J Lab Physicians 2011, 3:93-97.
• [23]Seema K, Ranjan Sen M, Upadhyay S, Bhattacharjee A: Dissemination of the New Delhi metallo-β-lactamase-1 (NDM-1) among Enterobacteriaceae in a tertiary referral hospital in north India. J Antimicrob Chemother 2011, 66:1646-1647.
• [24]Stuart CJ, Leverstein-Van Hall MA, The Dutch Working Party on the detection of highly resistant microorganisms: Guidelines for the phenotypic screening and the confirmation of carbapenemases in Enterobacteriaceae. Int J Antimicrob Agents 2010, 36:205-210.
• [25]Miriagou V, Cornaglia G, Edelstein M, Galani I, Giske CG, Gniadkowski M, Malamou-Lada E, Martinez-Martinez L, Navarro F, Nordmann P, Peixe L, Pournaras S, Rossolini GM, Tsakris A, Vatopoulos A, Canton R: Acquired carbapenemases in Gram-negative bacterial pathogens: detection and surveillance issues. Clin Microbiol Infect 2010, 16:112-122.
• [26]Nordmann P, Poirel L, Dortet L: Rapid detection of carbapenemase-producing Enterobacteriaceae. Emerg Infect Dis 2012, 18:1503-1507.
• [27]Maltezou HC: Metallo-beta-lactamases in Gram-negative bacteria: introducing the era of pan-resistance. Int J Antimicrob Agents 2009, 33:405.e1-7.
• [28]Al-Qadheeb NS, Althawadi S, Alkhalaf A, Hosaini S, Alrajhi AA: Evolution of tigecycline resistance in Klebsiella pneumoniae in a single patient. Ann Saudi Med 2010, 30:404-407.
• [29]Kanj SS, Kanafani ZA: Current concepts in the antimicrobial therapy against resistant gram-negative organisms: extended-spectrum beta-lactamase-producing Enterobacteriaceae, carbapenem resistant Enterobacteriaceae, and multidrug-resistant Pseudomonas aeruginosa. Mayo Clin Proc 2011, 86:250-259.