Phenotypic methods for detection of metallo-β-lactamase (MBL) production in multidrug resistant gram negative bacilli – comparative study


  • Umesh Santlal Hassani Department of Microbiology, NKP Salve Institute of Medical Sciences and Research Center and Lata Mangeshkar Hospital, Hingna MIDC, Digdoh Hills, Nagpur, Maharashtra
  • Rashmi Mahalle Department of Microbiology, NKP Salve Institute of Medical Sciences and Research Center and Lata Mangeshkar Hospital, Hingna MIDC, Digdoh Hills, Nagpur, Maharashtra



Multidrug resistant, Metallo betalactamase, Hodge test


Introduction: One of the most common mechanism of resistance of bacterial pathogens to β-lactam antibiotics is production of β-lactamase, there are different types of Beta lactamases, which are expressed by drug resistant gram-negative bacteria. Carbapenemases (Metallo beta lactamases/MBL) are the β-lactamases with the widest spectrum of activity. Early detection of MBL-producing organisms is crucial to establish appropriate antimicrobial therapy and to prevent their interhospital and intrahospital dissemination. Several phenotypic methods are available for the detection of MBL producing bacteria. As there is no standardized method present study was done to screen MDR gram negative bacilli isolated from clinical samples for MBL-production by a low cost, convenient and sensitive procedure. Methods: All non-duplicate MDR gram negative isolates obtained from various clinical samples were screened for carbapenam resistance. All carbapenam resistant bacteria were screened for production of MBL by 3 phenotypic tests (Double disc synergy, combined disk test, Hodge test). The results were compared and analyzed on the basis of results obtained by E test.  Results: During Study Period, 988 non duplicate gram negative bacilli were isolated, 70.64% (698) were multi drug resistant. Amongst Total number of MDR Isolates to carbapenam resistance was seen in 62(9.28%). These 62 isolates that were resistant to carbapenam were tested for MBL production. 54 (87%) of these 62 isolates showed MBL production by disc potentiation test whereas 41 isolates (66%) gave positive result by DDST. By Modified Hodge test, out of 62, 48 isolates (77.4%) were MBL positive. Compared to E- test, the Sensitivity Specificity and Accuracy for Disc potentiation test was 90%,100% and 90.32%, for Modified Hodge test was 80%,100% and 80.6% and for Double disc synergy test was 68.3%,100% and 69.3%. Conclusion: In our study, in comparison to MBL E test, disc potentiation test is more sensitive than double disc synergy test and Modified Hodge test for detection of MBL phenotypically


Download data is not yet available.


Pitout JD, Laupland KB. Extended-spectrum beta-lactamase-producing Enterobacteriaceae: an emerging public-health concern. Lancet Infect Dis 8, 159-66 (2008)

McGowan JE. Resistance in nonfermenting gram-negative bacteria: multidrug resistance to the maximum. Am J Med 119, S29-36; discussion S62-70 (2006)

Queenan AM, Bush K. Carbapenemases: the versatile beta-lactamases. Clin Microbiol Rev 20, 440-58, table of contents (2007)

Tato M, Coque TM, Ruíz-Garbajosa P, Pintado V, Cobo J, Sader HS, Jones RN, Baquero F, Cantón R. Complex clonal and plasmid epidemiology in the first outbreak of Enterobacteriaceae infection involving VIM-1 metallo-beta-lactamase in Spain: toward endemicity. Clin Infect Dis 45, 1171-8 (2007)

Qu TT, Zhang JL, Wang J, Tao J, Yu YS, Chen YG, Zhou JY, Li LJ. Evaluation of phenotypic tests for detection of metallo-beta-lactamase-producing Pseudomonas aeruginosa strains in China. J Clin Microbiol 47, 1136-42 (2009)

Magiorakos AP, Srinivasan A, Carey RB, Carmeli Y, Falagas ME, Giske CG, Harbarth S, Hindler JF, Kahlmeter G, Olsson-Liljequist B, Paterson DL, Rice LB, Stelling J, Struelens MJ, Vatopoulos A, Weber JT, Monnet DL. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect 18, 268-81 (2012)

Yong D, Lee K, Yum JH, Shin HB, Rossolini GM, Chong Y. Imipenem-EDTA disk method for differentiation of metallo-beta-lactamase-producing clinical isolates of Pseudomonas spp. and Acinetobacter spp. J Clin Microbiol 40, 3798-801 (2002)

Lee K, Chong Y, Shin HB, Kim YA, Yong D, Yum JH. Modified Hodge and EDTA-disk synergy tests to screen metallo-beta-lactamase-producing strains of Pseudomonas and Acinetobacter species. Clin Microbiol Infect 7, 88-91 (2001)

Walsh TR, Bolmström A, Qwärnström A, Gales A. Evaluation of a new Etest for detecting metallo-beta-lactamases in routine clinical testing. J Clin Microbiol 40, 2755-9 (2002)

Nordmann P, Cuzon G, Naas T. The real threat of Klebsiella pneumoniae carbapenemase-producing bacteria. Lancet Infect Dis 9, 228-36 (2009)

Kaleem F, Usman J, Hassan A, Khan A. Frequency and susceptibility pattern of metallo-beta-lactamase producers in a hospital in Pakistan. J Infect Dev Ctries 4, 810-3 (2010)

Cornaglia G, Akova M, Amicosante G, Cantón R, Cauda R, Docquier JD, Edelstein M, Frère JM, Fuzi M, Galleni M, Giamarellou H, Gniadkowski M, Koncan R, Libisch B, Luzzaro F, Miriagou V, Navarro F, Nordmann P, Pagani L, Peixe L, Poirel L, Souli M, Tacconelli E, Vatopoulos A, Rossolini GM. Metallo-beta-lactamases as emerging resistance determinants in Gram-negative pathogens: open issues. Int J Antimicrob Agents 29, 380-8 (2007)

Walsh TR, Toleman MA, Poirel L, Nordmann P. Metallo-beta-lactamases: the quiet before the storm. Clin Microbiol Rev 18, 306-25 (2005)

Walia K, Madhumathi J, Veeraraghavan B, Chakrabarti A, Kapil A, Ray P, Singh H, Sistla S, Ohri VC. Establishing Antimicrobial Resistance Surveillance & Research Network in India: Journey so far. Indian J Med Res 149, 164-79 (2019)

Siwakoti S, Subedi A, Sharma A, Baral R, Bhattarai NR, Khanal B. Incidence and outcomes of multidrug-resistant gram-negative bacteria infections in intensive care unit from Nepal- a prospective cohort study. Antimicrob Resist Infect Control 7, 114 (2018)

Saharman YR, Karuniawati A, Severin JA, Verbrugh HA. Infections and antimicrobial resistance in intensive care units in lower-middle income countries: a scoping review. Antimicrob Resist Infect Control 10, 22 (2021)

World Health Organization. Prioritization of pathogens to guide discovery, research and development of new antibiotics for drug-resistant bacterial infections, including tuberculosis. itiza tion-ofpatho gens/en/ (‎2017)‎

Mogasale VV, Saldanha P, Pai V, Rekha PD, Mogasale V. A descriptive analysis of antimicrobial resistance patterns of WHO priority pathogens isolated in children from a tertiary care hospital in India. Sci Rep 11, 5116 (2021)

Veeraraghavan B, Jesudason MR, Prakasah JAJ, Anandan S, Sahni RD, Pragasam AK, Bakthavatchalam YD, Selvakumar RJ, Dhole TN, Rodrigues C, Roy I, Joshi S, Chaudhuri BN, Chitnis DS. Antimicrobial susceptibility profiles of gram-negative bacteria causing infections collected across India during 2014-2016: Study for monitoring antimicrobial resistance trend report. Indian J Med Microbiol 36, 32-6 (2018)

Thacker N, Pereira N, Banavali SD, Narula G, Vora T, Chinnaswamy G, Prasad M, Kelkar R, Biswas S, Arora B. Epidemiology of blood stream infections in pediatric patients at a Tertiary Care Cancer Centre. Indian J Cancer 51, 438-41 (2014)

Sachdeva R, Sharma B, Sharma R. Evaluation of different phenotypic tests for detection of metallo-β-lactamases in imipenem-resistant Pseudomonas aeruginosa. J Lab Physicians 9, 249-53 (2017)

Behera B, Mathur P, Das A, Kapil A, Sharma V. An evaluation of four different phenotypic techniques for detection of metallo-beta-lactamase producing Pseudomonas aeruginosa. Indian J Med Microbiol 26, 233-7 (2008)

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 44, 3139-44 (2006)

Galani I, Rekatsina PD, Hatzaki D, Plachouras D, Souli M, Giamarellou H. Evaluation of different laboratory tests for the detection of metallo-beta-lactamase production in Enterobacteriaceae. J Antimicrob Chemother 61, 548-53 (2008)

Picão RC, Andrade SS, Nicoletti AG, Campana EH, Moraes GC, Mendes RE, Gales AC. Metallo-beta-lactamase detection: comparative evaluation of double-disk synergy versus combined disk tests for IMP-, GIM-, SIM-, SPM-, or VIM-producing isolates. J Clin Microbiol 46, 2028-37 (2008)

Pandya N, Oza S, Mehta S, Kikani K, Joshi P. Evaluation of various methods for detection of metallo-β-lactamase (MBL) production in gram negative bacilli. International Journal of Biological & Medical Research 2, 775-777 (2011).



How to Cite

Hassani, U. S. ., & Mahalle, R. . (2023). Phenotypic methods for detection of metallo-β-lactamase (MBL) production in multidrug resistant gram negative bacilli – comparative study. Journal of Applied Pharmaceutical Research, 11(3), 29-35.