Phenotypic and Genotypic Identification of Metallobetalactamase Genes in Resistant Enterobacteriaceae Isolated from Medical Centers in Isfahan
Novelty in Biomedicine,
Vol. 12 No. 1 (2024),
30 January 2024
,
Page 31- 42
Abstract
Background: The emergence of resistant Enterobacteriaceae and the abundance of antibiotic-resistant genes is one of the major problems of the global health system. The present study aimed to determine the phenotypic and genotypic expression levels of metallobetalactamase coding genes (blaVIM, blaNDM, blaIMP, blaSIM, blaSPM, and blaGIM) in Enterobacteriaceae isolates (Escherichia, Enterobacter, Citrobacter, Klebsiella, and Serratia) from patients referred to the clinical centers in Isfahan city and typing of these isolates.
Materials and Methods: Enterobacteriaceae isolates were identified and isolated after sample collection. Antibiotic sensitivity pattern was investigated by disk diffusion method. MIC was performed in carbapenem-resistant isolates by the E-test method, and the frequency of strains with multidrug resistance was determined. The presence of metallobetalactamase genes was investigated phenotypically using a combined disk test and modified Hodge test. The genotypic identification of the above genes was done by PCR and sequencing techniques. Finally, PCR based on the sequence of repetitive elements was performed for molecular typing of metallobetalactamase-producing Enterobacteriaceae.
Results: In the present study, 580 isolates of Enterobacteriaceae were isolated by examining 3500 samples. Klebsiella and Escherichia were the most common isolates, and the frequency of MDR was 60% in Klebsiella and 59.53% in Escherichia. Moreover, MIC results showed that 33.7% Klebsiella, 4.1% Escherichia, 5.7% Enterobacter, 3.5% Citrobacter, and 5.5% Serratia were resistant to carbapenems. Frequency of isolates with multidrug resistance in Escherichia (MDR 59.53% and XDR 1.5%), Klebsiella (MDR 60%, XDR% 3 and PDR 0.8%), Enterobacter (MDR 44%), Citrobacter (MDR 53.5%) and Serratia (MDR 55.5%) were reported. Metallobetalactamase production was confirmed by phenotypic analysis in Escherichia (1.8%) and Klebsiella (10.4%). Genotypic tests showed that blaSIM, blaSPM, and blaGIM genes were absent in any Enterobacteriaceae isolates. The presence of blaVIM, blaIMP, and blaNDM genes was confirmed in 6.2% of Klebsiella isolates and 1.3% of Escherichia isolates. The frequency of detected metallobetalactamase genes in Klebsiella and Escherichia isolates was 4.58% and 1.39% for blaVIM, 0.83% and 1.39% for blaIMP and 0.83% and 1.39% for blaNDM. The rep-PCR results showed that 11 metallobetalactamase-producing Klebsiella isolates are in 4 main groups, and 9 Escherichia isolates and 4 Enterobacter isolates are classified in two main clusters.
Conclusion: The present study shows the prevalence of Klebsiella and Escherichia isolates and their resistance to metallobetalactamase-producing Enterobacteriaceae. These genes in the horizontal transfer of antibiotic resistance identification of metallobetalactamase-producing isolates in clinical environments are essential to reduce the spread of antibiotic resistance. The high homology of resistant isolates of Enterobacteriaceae inclinical samples indicates the high power of these genotypes in causing infection in hospitalized patients, which can play an important role in increasing antibiotic resistance.
- Enterobacteriaceae
- Carbapenem, Metallobetalactamase enzyme
- Phenotype
- Genotype
How to Cite
References
Shahbazi S, Sabzi S, Goodarzi NN, Fereshteh S, Bolourchi N, Mirzaie B, et al. Identification of novel putative immunogenic targets and construction of a multi-epitope vaccine against multidrug-resistant Corynebacterium jeikeium using reverse vaccinology approach. Microbial Pathogenesis. 2022;164:105425.
Shahbazi S, Karam MRA, Habibi M, Talebi A, Bouzari S. Distribution of extended-spectrum β-lactam, quinolone and carbapenem resistance genes, and genetic diversity among uropathogenic Escherichia coli isolates in Tehran, Iran. Journal of global antimicrobial resistance. 2018;14:118-25.
Moghadam MT, Chegini Z, Norouzi A, Dousari AS, Shariati AJCpb. Three-decade failure to the eradication of refractory Helicobacter pylori infection and recent efforts to eradicate the infection. 2021;22(7):945-59.
Shahbazi R, Alebouyeh M, Shahkolahi S, Shahbazi S, Hossainpour H, Salmanzadeh-Ahrabi S. Molecular study on virulence and resistance genes of ST131 clone (uropathogenic/enteropathogenic Escherichia coli) hybrids in children. Future Microbiology. 2023;18(18):1353-61..
Shivaee A, Meskini M, Shahbazi S, Zargar M. Prevalence of flmA, flmH, mrkA, ecpA, and mrkD virulence genes affecting biofilm formation in clinical isolates of K. pneumonia. KAUMS Journal (FEYZ). 2019;23(2):168-76.
Shivaee A, Shahbazi S, Soltani A, Ahadi E. Evaluation of the prevalence of broad-spectrum beta-lactamases (ESBLs) and carbapenemase genes in Klebsiella pneumoniae strains isolated from burn wounds in patients referred to Shahid Motahari Hospital in Tehran. Med Sci J Islamic Azad Univ. 2019;29(3).
Pascale R, Giannella M, Bartoletti M, Viale P, Pea F. Use of meropenem in treating carbapenem-resistant Enterobacteriaceae infections. Expert Review of Anti-infective Therapy. 2019;17(10):819-27.
Ma J, Song X, Li M, Yu Z, Cheng W, Yu Z, et al. Global spread of carbapenem-resistant Enterobacteriaceae: Epidemiological features, resistance mechanisms, detection and therapy. Microbiological Research. 2022:127249.
Shahkolahi S, Shakibnia P, Shahbazi S, Sabzi S, Badmasti F, Asadi Karam MR, et al. Detection of ESBL and AmpC producing Klebsiella pneumoniae ST11 and ST147 from urinary tract infections in Iran. Acta Microbiologica et Immunologica Hungarica. 2022;69(4):303-13.
Shahbazi S, Habibi M, Badmasti F, Sabzi S, Farokhi M, Karam MRA. Design and fabrication of a vaccine candidate based on rOmpA from Klebsiella pneumoniae encapsulated in silk fibroin-sodium alginate nanoparticles against pneumonia infection. International Immunopharmacology. 2023;125:111171.
Goodarzi NN, Fereshteh S, Sabzi S, Shahbazi S, Badmasti F. Construction of a chimeric FliC including epitopes of OmpA and OmpK36 as a multi-epitope vaccine against Klebsiella pneumonia. Health Biothechnology and Biopharma. 2021;5:44-60.
Nelson RG, Rosowsky A. Dicyclic and tricyclic diaminopyrimidine derivatives as potent inhibitors of cryptosporidium parvum dihydrofolate reductase: structure-activity and structure-selectivity correlations. Antimicrobial Agents and Chemotherapy. 2002;46(3):940.
Moghadam M, Shariati A, Mirkalantari S, Karmostaji AJNM, Infections N. The complex genetic region conferring transferable antibiotic resistance in multidrug-resistant and extremely drug-resistant Klebsiella pneumoniae clinical isolates. 2020;36:100693.
Cheng Y, Wang C-y, Li Z-r, Pan Y, Liu M-b, Jiao Z. Can population pharmacokinetics of antibiotics be extrapolated? Implications of external evaluations. Clinical Pharmacokinetics. 2021;60:53-68.
Mohebi S, Hossieni Nave H, Norouzi A, Kandehkar Gharaman M, Taati Moghadam MJJoMUoMS. Detection of extended spectrum beta lactamases on class I integron in Escherichia coli isolated from clinical samples. 2016;26(138):66-76.
Moghadam MT, Mojtahedi A, Moghaddam MM, Fasihi-Ramandi M, Mirnejad RJAm, biotechnology. Rescuing humanity by antimicrobial peptides against colistin-resistant bacteria. 2022;106(11):3879-93.
Datta P, Gupta V, Garg S, Chander J. Phenotypic method for differentiation of carbapenemases in Enterobacteriaceae: Study from north India. Indian Journal of Pathology and Microbiology. 2012;55(3):357.
Kollenda H, Frickmann H, Helal RB, Wiemer DF, Naija H, El Asli MS, et al. Screening for carbapenemases in ertapenem-resistant Enterobacteriaceae collected at a Tunisian hospital between 2014 and 2018. European Journal of Microbiology and Immunology. 2019;9(1):9-13.
Taati Moghadam M, Hossieni Nave H, Mohebi S, Norouzi AJIJoMM. The evaluation of connection between integrons class I and II and ESBL-producing and Non-ESBL Klebsiella pneumoniae isolated from clinical samples, Kerman. 2016;10(4):1-9.
Mahon CR, Lehman DC. Textbook of diagnostic microbiology-e-book: Elsevier Health Sciences; 2022.
Li S, Duan X, Peng Y, Rui Y. Molecular characteristics of carbapenem-resistant Acinetobacter spp. from clinical infection samples and fecal survey samples in Southern China. BMC infectious diseases. 2019;19(1):1-12.
Bartolini A, Frasson I, Cavallaro A, Richter SN, Palù G. Comparison of phenotypic methods for the detection of carbapenem non-susceptible Enterobacteriaceae. Gut pathogens. 2014;6(1):1-7.
Thomson KS, Moland ES. Version 2000: the new β-lactamases of Gram-negative bacteria at the dawn of the new millennium. Microbes and infection. 2000;2(10):1225-35.
Simbolo M, Gottardi M, Corbo V, Fassan M, Mafficini A, Malpeli G, et al. DNA qualification workflow for next generation sequencing of histopathological samples. PloS one. 2013;8(6):e62692.
Hujer KM, Hujer AM, Hulten EA, Bajaksouzian S, Adams JM, Donskey CJ, et al. Analysis of antibiotic resistance genes in multidrug-resistant Acinetobacter sp. isolates from military and civilian patients treated at the Walter Reed Army Medical Center. Antimicrobial agents and chemotherapy. 2006;50(12):4114-23.
Lü Y, Liang H, Zhang W, Liu J, Wang S, Hu H. A novel bla SIM-1-carrying megaplasmid pSIM-1-BJ01 isolated from clinical Klebsiella pneumonia. bioRxiv. 2019:519082.
Norouzi Bazgir Z, Ahanjan M, Hashemi Soteh MB, Reza Goli H. Prevalence of IMP and SPM Genes in Clinical Isolatesof Carbapenem Resistant Acinetobacter baumannii in Educational Hospitals of Sari, Iran. Journal of Mazandaran University of Medical Sciences. 2019;29(174):30-41.
Bi R, Kong Z, Qian H, Jiang F, Kang H, Gu B, et al. High prevalence of bla NDM variants among carbapenem-resistant Escherichia coli in Northern Jiangsu Province, China. Frontiers in microbiology. 2018;9:2704.
Tarashi S, Goudarzi H, Erfanimanesh S, Pormohammad A, Hashemi A. Phenotypic and molecular detection of metallo-beta-lactamase genes among imipenem resistant Pseudomonas aeruginosa and Acinetobacter baumannii strains isolated from patients with burn injuries. Archives of Clinical Infectious Diseases. 2016;11(4).
Avcioglu NH, Bilkay IS. Antibiotic resistance, multidrug resistance and Enterobacterial repetitive intergenic consensus polymerase chain reaction profiles of clinically important Klebsiella species. Asian Biomedicine. 2016;10(1):41-7.
Kholodok G, Alekseeva I, Strel'nikova N, Kozlov V. Colonization properties of opportunistic bacteria isolated from children with pneumonia. Zhurnal Mikrobiologii, Epidemiologii i Immunobiologii. 2014(2):17-25.
Mirbagheri SZ, Meshkat Z, Naderinasab M, Rostami S, Nabavinia MS, Rahmati M. Study on imipenem resistance and prevalence of blaVIM1 and blaVIM2 metallo-beta lactamases among clinical isolates of Pseudomonas aeruginosa from Mashhad, Northeast of Iran. Iranian Journal of Microbiology. 2015;7(2):72.
Tilahun M, Kassa Y, Gedefie A, Ashagire M. Emerging carbapenem-resistant Enterobacteriaceae infection, its epidemiology and novel treatment options: a review. Infection and Drug Resistance. 2021:4363-74.
Lutgring JD, Limbago BM. The problem of carbapenemase-producing-carbapenem-resistant-Enterobacteriaceae detection. Journal of clinical microbiology. 2016;54(3):529-34.
Tzouvelekis L, Markogiannakis A, Psichogiou M, Tassios P, Daikos G. Carbapenemases in Klebsiella pneumoniae and other Enterobacteriaceae: an evolving crisis of global dimensions. Clinical microbiology reviews. 2012;25(4):682-707.
Japoni-Nejad A, Ghaznavi-Rad E, Van Belkum A. Characterization of plasmid-mediated AmpC and carbapenemases among Iranain nosocomial isolates of Klebsiella pneumoniae using phenotyping and genotyping methods. Osong public health and research perspectives. 2014;5(6):333-8.
Norouzi Bazgir Z, Mirzaei B, Haghshenas MR, Goli HR, Shafaie E. Multidrug Resistant Citrobacter freundii Isolates in a Burn Hospital in Northeast of Iran: A Single-Center Cross-sectional Study. Research in Molecular Medicine. 2020;8(2):63-70.
Peymani A, Najafipour R. Multidrug resistance in Pseudomonas aeruginosa and Enterobacter cloacae isolated from intensive care units of Qazvin and Tehran hospitals. Journal of Clinical Research in Paramedical Sciences. 2014;3(1).
Sanchez GV, Master RN, Clark RB, Fyyaz M, Duvvuri P, Ekta G, et al. Klebsiella pneumoniae antimicrobial drug resistance, United States, 1998–2010. Emerging infectious diseases. 2013;19(1):133.
Babaie Kasmaei Z, Mozafari N, Forohesh Tehrani H, Arashkia A, Mahdavi S, Bahrami A. Determine the antimicrobial susceptibility of E. coli with multiple drug resistance in outpatients with urinary tract infection in Tehran. Iran J Med Microbiol. 2012;6(9):37-44.
Bhatt P, Tandel K, Shete V, Rathi K. Burden of extensively drug-resistant and pandrug-resistant Gram-negative bacteria at a tertiary-care centre. New microbes and New infections. 2015;8:166-70.
Srinivasan R, Karaoz U, Volegova M, MacKichan J, Kato-Maeda M, Miller S, et al. Use of 16S rRNA gene for identification of a broad range of clinically relevant bacterial pathogens. PloS one. 2015;10(2):e0117617.
Hashemi A, Tarashi S, Erfanimanesh SJAV. Epidemiology of antibiotic resistance and MBL genes among Pseudomonas aeruginosa and Acinetobacter baumannii clinical strains isolated from burnt patients in Iran: a systematic review. 2016;14(1):40-3.
Dortet L, Poirel L, Nordmann P. Worldwide dissemination of the NDM-type carbapenemases in Gram-negative bacteria. BioMed research international. 2014;2014.
Latifi B, Tajbakhsh S, Askari A, Yousefi F. Phenotypic and genotypic characterization of carbapenemase-producing Klebsiella pneumoniae clinical isolates in Bushehr province, Iran. Gene Reports. 2020;21:100932.
Ghadiri H, Vaez H, Razavi-Azarkhiavi K, Rezaee R, Haji-Noormohammadi M, Rahimi AA, et al. Prevalence and antibiotic susceptibility patterns of Extended-Spectrum ß-Lactamase and Metallo-ß-Lactamase–producing uropathogenic Escherichia coli isolates. Laboratory medicine. 2014;45(4):291-6.
Ratkai C, Quinteira S, Grosso F, Monteiro N, Nagy E, Peixe L. Controlling for false positives: interpreting MBL Etest and MBL combined disc test for the detection of metallo-β-lactamases. Journal of antimicrobial chemotherapy. 2009;64(3):657-8.
Shams S, Hashemi A, Kermani S, Esmkhani M, Tarashi S. Determination of Extended-spectrum Beta-lactamase Genes (blaTEM, blaSHV, blaCTX-M) in Escherichia coli Strains Isolated from Clinical Specimenes in Ali-Ibne Abi Talib Hospital in Qom, Iran. 2016.
Gondal AJ, Choudhry N, Bukhari H, Rizvi Z, Jahan S, Yasmin N. Estimation, Evaluation and Characterization of Carbapenem Resistance Burden from a Tertiary Care Hospital, Pakistan. Antibiotics. 2023;12(3):525.
Kakian F, Naderi K, Rezaei MH, Validi M, Zamanzad B, Gholipour A. Identification of VIM and IMP genes and metallo-betalactamase enzymes in Escherichia coli isolates by molecular and phenotypic methods in shahrekord educational hospitals. Journal of Shahrekord University of Medical Sciences. 2020;22(1):46-52.
Findlay J, Poirel L, Kessler J, Kronenberg A, Nordmann P. New Delhi Metallo-β-Lactamase–Producing Enterobacterales Bacteria, Switzerland, 2019–2020. Emerging infectious diseases. 2021;27(10):2628.
Khalid S, Ahmad N, Ali SM, Khan AU. Outbreak of efficiently transferred carbapenem-resistant blaNDM-producing gram-negative bacilli isolated from neonatal intensive care unit of an Indian hospital. Microbial Drug Resistance. 2020;26(3):284-9.
Davari N, Khashei R, Pourabbas B, Nikbin VS, Zand F. High frequency of carbapenem-resistant Enterobacteriaceae fecal carriage among ICU hospitalized patients from Southern Iran. Iranian Journal of Basic Medical Sciences. 2022;25(12):1416.
Solgi H, Nematzadeh S, Giske CG, Badmasti F, Westerlund F, Lin Y-L, et al. Molecular epidemiology of OXA-48 and NDM-1 producing Enterobacterales species at a University Hospital in Tehran, Iran, between 2015 and 2016. Frontiers in microbiology. 2020;11:936.
Shams S, Hashemi A, Esmkhani M, Kermani S, Shams E, Piccirillo A. Imipenem resistance in clinical Escherichia coli from Qom, Iran. BMC Research Notes. 2018;11:1-5.
Castanheira M, Mendes RE, Woosley LN, Jones RN. Trends in carbapenemase-producing Escherichia coli and Klebsiella spp. from Europe and the Americas: report from the SENTRY antimicrobial surveillance programme (2007–09). Journal of antimicrobial chemotherapy. 2011;66(6):1409-11.
Ventola CL. The antibiotic resistance crisis: part 1: causes and threats. Pharmacy and therapeutics. 2015;40(4):277.
Seiffert SN, Marschall J, Perreten V, Carattoli A, Furrer H, Endimiani A. Emergence of Klebsiella pneumoniae co-producing NDM-1, OXA-48, CTX-M-15, CMY-16, QnrA and ArmA in Switzerland. International journal of antimicrobial agents. 2014;44(3):260-2.
Messaoudi A, Haenni M, Mansour W, Saras E, Bel Haj Khalifa A, Chaouch C, et al. ST147 NDM-1-producing Klebsiella pneumoniae spread in two Tunisian hospitals. Journal of Antimicrobial Chemotherapy. 2016;72(1):315-6.
Ranjbar R, Tabatabaee A, Behzadi P, Kheiri R. Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) genotyping of Escherichia coli strains isolated from different animal stool specimens. Iranian journal of pathology. 2017;12(1):25.
Asgharzadeh S, Golmoradi Zadeh R, Taati Moghadam M, Farahani Eraghiye H, Sadeghi Kalani B, Masjedian Jazi F, et al. Distribution and expression of virulence genes (hlyA, sat) and genotyping of Escherichia coli O25b/ST131 by multi-locus variable number tandem repeat analysis in Tehran, Iran. 2022;69(4):314-22.
Khoshbayan A, Golmoradi Zadeh R, Taati Moghadam M, Mirkalantari S, Darbandi AJAoCM, Antimicrobials. Molecular determination of O25b/ST131 clone type among extended spectrum β-lactamases production Escherichia coli recovering from urinary tract infection isolates. 2022;21(1):35.
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