DOI: https://dx.doi.org/10.18203/2320-6012.ijrms20221782
Published: 2022-06-28

Comparison of combined disc synergy test and double disc synergy test for phenotypic detection of metallo-ß-lactamase among the clinical isolates of gram-negative bacilli

Rifat Asma Chowdhury, Sabrina Sharmin, Abu Hena M. Saiful Karim Chowdhury, Nasima Akter, Shakeel Ahmed, Gulshan Nigar Choudhury

Abstract


Background: Metallo-β-lactamases (MBL) have a wide spectrum of activity and they confer a higher level of resistance to all β-lactams antibiotics including Carbapenem. The active site in MBLs contains either 1 or 2 Zn2+ ions for their catalytic mechanism. All MBLs share a common feature of being inhibited by EDTA. Metallo-β-lactamase-producing gram-negative bacteria are the most important nosocomial pathogens. The present study was conducted to detect Metallo-β-lactamase (MBL) production in gram-negative bacilli by Combined Disc Synergy Test (CDST) and Double Disc Synergy Test (DDST) with 0.1M EDTA as a chelator and to see their antibiotic susceptibility pattern of them.

Methods: The cross-sectional observational study was carried out in the Department of Microbiology, Chittagong Medical College, during the period of July 2015 to June 2016. Samples were collected from patients admitted to CMCH. Standard Microbiological procedures and biochemical tests were carried out for the isolation and identification of MBL. SPSS software is used for data analysis.

Results: When 66 screening positive MBL isolates were subjected to the phenotypic confirmatory test CDST detected 50 (25.4%) and DDST detected 48 (24.4%) as MBL producers. Among these isolates, we found Acinetobacter spp. 7 (100%), as the leading MBL producer followed by Pseudomonas spp. 16 (32.6%), E. coli 10 (20%) and Klebsiella spp. 15 (17.4%).

Conclusions: In a laboratory where multiplex PCR molecular set-up is not available CDST and DDST are convenient phenotypic methods and can be implemented in routine microbiological laboratories as well as in primary health care setup for daily application to monitor the production of MBLs.


Keywords


Microbiological, Antibiotics, Pathogen, Bacteria, MBL, Synergy

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References


Walsh TR. Emerging Carbapenemases: a global perspective. International Journal of Antimicrobial Agents. 2011;508-14.

Peleg AY, Franklin C, Bell MJ, Spelman D. Dissemination of the Metallo-β-Lactamase Gene blaIMP-4 among Gram-Negative Pathogens in a Clinical Setting in Australia. Clinical Infectious Diseases. 2005;41:1549-56.

Watanabe M, Iyobe S, Inoue M, Mitsuhashi S. Transferable imipenem resistance in Pseudomonas aeruginosa. Antimicrobial agents and chemotherapy. 1991;35(1):147-51.

Cornaglia G, H Giamarellou, Rossolini GM. Metallo-β-lactamases last frontier for β-lactams? International Business Review. 2011;11(5):381-93.

Kumar SH, Baveja SM & Gore MA. Prevalance and risk factors of metallo beta lactamases producing Pseudomonas aeruginosa and Acinetobacter species in Burns and Surgical wards in a tertiary care hospital. Journal of Labratory Physicians. 2012;4(1):39-42.

Anwar S. Phenotypic detection of Metallo-β-Lactamases among the Clinical isolates of Pseudomonas spp. and Acinetobacter spp., M.Phil Thesis, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh. 2010.

Arakawa Y, Shibata N, Shibayama K, Kurokawa H, Yagi T, Fujiwara H, Goto M. Convenient Test for Screening Metallo-β-Lactamase Producing Gram-Negative Bacteria by Using Thiol Compound. Journal of Clinical Microbiology. 2000;38(1):40-3.

Yong D, Lee K, Yum JH, Shin HB, Rossolini GM, Chong Y. Imipenem-EDTA Disk Method for Differentiation of Metallo-β -lactamases- producing Clinical Isolates of Pseudomonas spp. and Acinetobacter spp. Journal of Clinical Microbiology. 2002;40(10):3798-801.

Franklin C, Liolios L & Peleg AY. Phenotypic Detection of Carbapenem-Suspectable Metallo-Beta-Lactamase Producing Gram-negative Bacilli in the Clinical Laboratory. Journal of Clinical Microbiology. 2006;44(9):3139-44.

Bashir D, Thoker MA, Fomda BA, Bashir G, Zahoor A, Toboli AS. Detection of Metallo-beta-lactamase (MBL) producing Pseudomonas aeruginosa at a Teritary Care Hospital in Kashmir. American Journal of Microbiology Research. 2011;5(2):164-72.

Rasheed JK, Kitchel B, Zhu W, Anderson KF, Clark NC, Ferraro MJ et al. New Delhi Metallo-β-Lactamase Producing Enterobacteriaceae United States. Emerging Infectious Diseases. 2013;19(6):870-80.

Struelens MJ, Monnet DL, Magiorakos AP, O’Conner FS. New Delhi metallo-beta-lactamase 1 producing Enterobacteriaceae: emergence and response in Europe, European Centre for Disease Prevention and Control (ECDC), Stockholm, Sweden. 2010;1-10.

Shamuzzaman SM. NDM1 producing new superbug bacteria: a threat to control infection. Bangladesh J Med Microbiol. 2011;5(1):1-2.

Nasrin T, Jilani SA, Barai L, Haq JA. Metallo-β-lactamase Producing Pseudomonas species in a Teriary care Hospital of Dhaka City. Bangladesh J Med Microbial. 2011;4(1):43-5.

Anwar S, Miah MRA, Saleh AA, Sattar H, Ahmed S. Simple Screening Test for The Detection of Metallo-β-lactamase (MBL) Production In Clinical Isolates Pseudomonas and Acinetobacter. Ibrahim Med Coll J. 2010;4(1):26-30.

Agarwal R, Sumana MN, Kishore A, Kulkarni M. Simple Method for Detection of Metallo β Lactamase among Gram-negative Isolates. Online journal of Health and Allied Science. 2015;14(3):1-3.

Wankhede SV, Iyer VS, Bharadwaj RS. The Study of MBL Producers in Gram-negative Isolates from ICUs and Wards. Indian Journal of Basic and Applied Medical Research. 2011;1(1):38-46.

Saini M, Mishra A, Gupta S. Prevalence of Carbapenem Resistance in Gram-negative Bacilli Isolates and Their Antimicrobial Susceptibility Pattern. Int J Med Res Prof. 2016;2(3):28-32.

Agarwal R, Sumana MN, Kishore A, Kulkarni M. Simple Method for Detection of Metallo β Lactamase among Gram-negative Isolates. Online journal of Health and Allied Science. 2015;14(3):1-3.

Patel DD, Rathod SD, Panchal PD, Chauhan B, Pethani JD, Shah PD. Comparison of Two Phenotypic methods, Prevalence and Antimicrobial Suspectibility profile of Metallo-β-Lactamase(MBL) Producing Gram-negative bacilli. International Journal of Scientific Research. 2013;2(7):3124.

Bradford PA. Extended-Spectrum β-Lactamases in the 21st Century: Characterization, Epidemiology and Detection of This important Resistance Threat. Clinical Microbiology Reviews. 2001;14(4):933-51.

Noyal MJC, Menezes G, Harish BN, Sujatha S, Parija SC. Simple screening tests for detection of carbapenemases inclinical isolates of non-fermentative gram-negative bacteria. Indian j Med Res. 2009;707-12.

Tellis R, Muralidharan S, Peter AI. Evaluation of three phenotypic methods for the detection of metallo-beta-lactamase production in non-fermenting gram-negative bacilli. International Journal of Biomedical And Advance research. 2013;4(5):299-305.