Login Register Cart 0
Generic placeholder image

Combinatorial Chemistry & High Throughput Screening

Editor-in-Chief

ISSN (Print): 1386-2073
ISSN (Online): 1875-5402

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Research Article

Detection of MCR-1 Gene in Multiple Drug Resistant Escherichia coli and Klebsiella pneumoniae in Human Clinical Samples from Peshawar, Pakistan

Author(s): Fareeha Hameed, Muhammad Asif Khan, Hazrat Bilal, Hafsah Muhammad and Tayyab Ur Rehman*

Volume 24, Issue 5, 2021

Published on: 14 September, 2020

Page: [737 - 742] Pages: 6

DOI: 10.2174/1386207323666200914100119

Price: $65

Abstract

Background: The presence of plasmid mediated mcr-1 gene in multidrug resistant Gram-negative bacteria poses a serious public health concern in today’s world.

Objective: The present study was aimed to detect the presence of plasmid mediated mcr-1 encoding resistance to colistin in multiple drug resistant (MDR) E. coli and K. pneumoniae isolates.

Methods: A total of 180 clinical isolates of E. coli (n=120) and K. pneumoniae (n=60) were isolated from different clinical specimens, i.e., urine, blood, stool and pus, from diagnostic labs of two major public sector tertiary care hospitals in Peshawar, Pakistan. MDR profile of these isolates was assessed through Kirby-Baur disc diffusion method. All isolates were screened for colistin resistance by dilution methods. Colistin resistant isolates were subjected to PCR for mcr-1 detection and confirmation was done by Sanger sequencing method.

Results: Overall, 83.3% (100/120) E. coli and 93.3% (56/60) K. pneumoniae were detected as MDR. Colistin resistance was found in 23.3% (28/120) E. coli and 40% (24/60) K. pneumoniae isolates, whereas mcr-1 gene was detected in 10 out of 52 colistin resistant isolates, including six E. coli and four K. pneumoniae isolates. Minimum inhibitory concentrations (MICs) of colistin in these ten mcr-1 positive isolates ranged from 4μg/ml to 16μg/ml. All mcr-1 positive isolates showed 99% sequence similarity when compared with other present sequences in GenBank.

Conclusion: Hence, our study confirms the presence of mcr-1 mediated colistin resistance in the studied area. Therefore, urgently larger scale surveillance studies are recommended to investigate prevalence of mcr-1 mediated colistin resistance and to prevent its further spread in the area.

Keywords: Multiple drug resistance, colistin resistance, mcr-1, Escherichia coli, Klebsiella pneumoniae, biosynthesis.

« Previous
[1]
Gonzales Escalante, E.; Yauri Condor, K.; Di Conza, J.A.; Gutkind, G.O. Phenotypic detection of plasmid-mediated colistin resistance in Enterobacteriaceae. J. Clin. Microbiol., 2020, 58(3), e01555-e19.
[PMID: 31801838]
[2]
Zafer, M.M.; El-Mahallawy, H.A.; Abdulhak, A.; Amin, M.A.; Al-Agamy, M.H.; Radwan, H.H. Emergence of colistin resistance in multidrug-resistant Klebsiella pneumoniae and Escherichia coli strains isolated from cancer patients. Ann. Clin. Microbiol. Antimicrob., 2019, 18(1), 40.
[http://dx.doi.org/10.1186/s12941-019-0339-4] [PMID: 31831019]
[3]
El-Sayed Ahmed, M.A.E.; Zhong, L-L.; Shen, C.; Yang, Y.; Doi, Y.; Tian, G-B. Colistin and its role in the Era of antibiotic resistance: an extended review (2000-2019). Emerg. Microbes Infect., 2020, 9(1), 868-885.
[http://dx.doi.org/10.1080/22221751.2020.1754133] [PMID: 32284036]
[4]
Doern, C.D.; Holfelder, M. Automation and design of the clinical microbiology laboratory.Manual of Clinical Microbiology, 11th ed; American Society of Microbiology, 2015, pp. 44-53.
[5]
Yu, Z.; Qin, W.; Lin, J.; Fang, S.; Qiu, J. Antibacterial mechanisms of polymyxin and bacterial resistance BioMed research international, 2015.
[http://dx.doi.org/10.1155/2015/679109]
[6]
Wang, X.; Quinn, P.J. Lipopolysaccharide: Biosynthetic pathway and structure modification. Prog. Lipid Res., 2010, 49(2), 97-107.
[http://dx.doi.org/10.1016/j.plipres.2009.06.002] [PMID: 19815028]
[7]
Liu, Y-Y.; Wang, Y.; Walsh, T.R.; Yi, L-X.; Zhang, R.; Spencer, J.; Doi, Y.; Tian, G.; Dong, B.; Huang, X.; Yu, L.F.; Gu, D.; Ren, H.; Chen, X.; Lv, L.; He, D.; Zhou, H.; Liang, Z.; Liu, J.H.; Shen, J. Emergence of plasmid-mediated colistin resistance mechanism MCR-1 in animals and human beings in China: a microbiological and molecular biological study. Lancet Infect. Dis., 2016, 16(2), 161-168.
[http://dx.doi.org/10.1016/S1473-3099(15)00424-7] [PMID: 26603172]
[8]
Newton-Foot, M.; Snyman, Y.; Maloba, M.R.B.; Whitelaw, A.C. Plasmid-mediated mcr-1 colistin resistance in Escherichia coli and Klebsiella spp. clinical isolates from the Western Cape region of South Africa. Antimicrob. Resist. Infect. Control, 2017, 6(1), 78.
[http://dx.doi.org/10.1186/s13756-017-0234-8] [PMID: 28785405]
[9]
Xavier, B.B.; Lammens, C.; Ruhal, R.; Kumar-Singh, S.; Butaye, P.; Goossens, H.; Malhotra-Kumar, S. Identification of a novel plasmid-mediated colistin-resistance gene, mcr-2, in Escherichia coli, Belgium, June 2016. Euro Surveill., 2016, 21(27), 30280.
[http://dx.doi.org/10.2807/1560-7917.ES.2016.21.27.30280] [PMID: 27416987]
[10]
Yin, W.; Li, H.; Shen, Y.; Liu, Z.; Wang, S.; Shen, Z.; Zhang, R.; Walsh, T.R.; Shen, J.; Wang, Y. Novel plasmid-mediated colistin resistance gene mcr-3 in Escherichia coli. MBio, 2017, 8(3), e00543-e17.
[http://dx.doi.org/10.1128/mBio.00543-17] [PMID: 28655818]
[11]
Carattoli, A.; Villa, L.; Feudi, C.; Curcio, L.; Orsini, S.; Luppi, A.; Pezzotti, G.; Magistrali, C.F. Novel plasmid-mediated colistin resistance mcr-4 gene in Salmonella and Escherichia coli, Italy 2013, Spain and Belgium, 2015 to 2016. Euro Surveill., 2017, 22(31), 30589.
[http://dx.doi.org/10.2807/1560-7917.ES.2017.22.31.30589] [PMID: 28797329]
[12]
Borowiak, M.; Fischer, J.; Hammerl, J.A.; Hendriksen, R.S.; Szabo, I.; Malorny, B. Identification of a novel transposon-associated phosphoethanolamine transferase gene, mcr-5, conferring colistin resistance in d-tartrate fermenting Salmonella enterica subsp. enterica serovar Paratyphi B. J. Antimicrob. Chemother., 2017, 72(12), 3317-3324.
[http://dx.doi.org/10.1093/jac/dkx327] [PMID: 28962028]
[13]
AbuOun, M.; Stubberfield, E.J.; Duggett, N.A.; Kirchner, M.; Dormer, L.; Nunez-Garcia, J.; Randall, L.P.; Lemma, F.; Crook, D.W.; Teale, C.; Smith, R.P.; Anjum, M.F. mcr-1 and mcr-2 variant genes identified in Moraxella species isolated from pigs in Great Britain from 2014 to 2015. J. Antimicrob. Chemother., 2017, 72(10), 2745-2749.
[http://dx.doi.org/10.1093/jac/dkx286] [PMID: 29091227]
[14]
Yang, Y-Q.; Li, Y-X.; Lei, C-W.; Zhang, A-Y.; Wang, H-N. Novel plasmid-mediated colistin resistance gene mcr-7.1 in Klebsiella pneumoniae. J. Antimicrob. Chemother., 2018, 73(7), 1791-1795.
[http://dx.doi.org/10.1093/jac/dky111] [PMID: 29912417]
[15]
Wang, X.; Wang, Y.; Zhou, Y.; Li, J.; Yin, W.; Wang, S.; Zhang, S.; Shen, J.; Shen, Z.; Wang, Y. Emergence of a novel mobile colistin resistance gene, mcr-8, in NDM-producing Klebsiella pneumoniae. Emerg. Microbes Infect., 2018, 7(1), 122.
[http://dx.doi.org/10.1038/s41426-018-0124-z] [PMID: 29970891]
[16]
Carroll, L.M.; Gaballa, A.; Guldimann, C.; Sullivan, G.; Henderson, L.O.; Wiedmann, M. Identification of novel mobilized colistin resistance gene mcr-9 in a multidrug-resistant, colistin-susceptible Salmonella enterica serotype typhimurium isolate. MBio, 2019, 10(3), e00853-e19.
[http://dx.doi.org/10.1128/mBio.00853-19] [PMID: 31064835]
[17]
Wang, C.; Feng, Y.; Liu, L.; Wei, L.; Kang, M.; Zong, Z. Identification of novel mobile colistin resistance gene mcr-10. Emerg. Microbes Infect., 2020, 9(1), 508-516.
[http://dx.doi.org/10.1080/22221751.2020.1732231] [PMID: 32116151]
[18]
Hudzicki, J. Kirby-Bauer disk diffusion susceptibility test protocol 2009.
[19]
Clinical; Institute, L. S., Performance Standards for Antimicrobial Testing: Twenty-Sixth Informational Supplement M100-S26 2016,
[20]
Wiegand, I.; Hilpert, K.; Hancock, R.E. Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat. Protoc., 2008, 3(2), 163-175.
[http://dx.doi.org/10.1038/nprot.2007.521] [PMID: 18274517]
[21]
Testing, E.C.A.S. European Committee on Antimicrobial Susceptibility Testing Breakpoint tables for interpretation of MICs and zone diameters European Committee on Antimicrobial Susceptibility Testing Breakpoint tables for interpretation of MICs and zone diameters; Växjö, Sweden, 2015.
[22]
Feliciello, I.; Chinali, G. A modified alkaline lysis method for the preparation of highly purified plasmid DNA from Escherichia coli. Anal. Biochem., 1993, 212(2), 394-401.
[http://dx.doi.org/10.1006/abio.1993.1346] [PMID: 8214582]
[23]
Abrar, S.; Hussain, S.; Khan, R.A.; Ul Ain, N.; Haider, H.; Riaz, S. Prevalence of extended-spectrum-β-lactamase-producing Enterobacteriaceae: first systematic meta-analysis report from Pakistan. Antimicrob. Resist. Infect. Control, 2018, 7(1), 26.
[http://dx.doi.org/10.1186/s13756-018-0309-1] [PMID: 29484173]
[24]
Shaikh, B.T. Anti-microbial resistance in Pakistan: a public health issue. J. Ayub Med. Coll. Abbottabad, 2017, 29(2), 184-185.
[PMID: 28718227]
[25]
Nordmann, P.; Poirel, L. Plasmid-mediated colistin resistance: an additional antibiotic resistance menace. Clin. Microbiol. Infect., 2016, 22(5), 398-400.
[http://dx.doi.org/10.1016/j.cmi.2016.03.009] [PMID: 27021419]
[26]
Park, K. S.; Lee, J. H.; Park, M.; Ko, K. S.; Lee, S. H. How many mcr-1-harbouring bacteria were spreading geographically? 2017.
[27]
Goli, H.R.; Nahaei, M.R.; Ahangarzadeh Rezaee, M.; Hasani, A.; Samadi Kafil, H.; Aghazadeh, M. Emergence of colistin resistant Pseudomonas aeruginosa at Tabriz hospitals, Iran. Iran. J. Microbiol., 2016, 8(1), 62-69.
[PMID: 27092226]
[28]
Pragasam, A.K.; Shankar, C.; Veeraraghavan, B.; Biswas, I.; Nabarro, L.E.; Inbanathan, F.Y.; George, B.; Verghese, S. Molecular mechanisms of colistin resistance in Klebsiella pneumoniae causing bacteremia from India—a first report. Front. Microbiol., 2017, 7, 2135.
[http://dx.doi.org/10.3389/fmicb.2016.02135] [PMID: 28119670]
[29]
Dafopoulou, K.; Zarkotou, O.; Dimitroulia, E.; Hadjichristodoulou, C.; Gennimata, V.; Pournaras, S.; Tsakris, A. Comparative evaluation of colistin susceptibility testing methods among carbapenem-nonsusceptible Klebsiella pneumoniae and Acinetobacter baumannii clinical isolates. Antimicrob. Agents Chemother., 2015, 59(8), 4625-4630.
[http://dx.doi.org/10.1128/AAC.00868-15] [PMID: 26014928]
[30]
Cannatelli, A.; Giani, T.; Antonelli, A.; Principe, L.; Luzzaro, F.; Rossolini, G. M. First detection of the mcr-1 colistin resistance gene in Escherichia coli, Italy. Antimicrobial agents and chemotherapy, 2016.
[31]
Zurfluh, K.; Nüesch-Inderbinen, M.; Klumpp, J.; Poirel, L.; Nordmann, P.; Stephan, R. Key features of mcr-1-bearing plasmids from Escherichia coli isolated from humans and food. Antimicrob. Resist. Infect. Control, 2017, 6(1), 91.
[http://dx.doi.org/10.1186/s13756-017-0250-8] [PMID: 28878890]
[32]
Chan, W-S.; Au, C-H.; Ho, D.N.; Chan, T-L.; Ma, E.S-K.; Tang, B.S-F. Prospective study on human fecal carriage of Enterobacteriaceae possessing mcr-1 and mcr-2 genes in a regional hospital in Hong Kong. BMC Infect. Dis., 2018, 18(1), 81.
[http://dx.doi.org/10.1186/s12879-018-2987-y] [PMID: 29439654]
[33]
Berglund, B.; Hoang, N.T.B.; Tärnberg, M.; Le, N.K.; Welander, J.; Nilsson, M.; Khu, D.T.K.; Nilsson, L.E.; Olson, L.; Le, H.T.; Larsson, M.; Hanberger, H. Colistin- and carbapenem-resistant Klebsiella pneumoniae carrying mcr-1 and blaOXA-48 isolated at a paediatric hospital in Vietnam. J. Antimicrob. Chemother., 2018, 73(4), 1100-1102.
[http://dx.doi.org/10.1093/jac/dkx491] [PMID: 29253209]
[34]
Wise, M.G.; Estabrook, M.A.; Sahm, D.F.; Stone, G.G.; Kazmierczak, K.M. Prevalence of mcr-type genes among colistin-resistant Enterobacteriaceae collected in 2014-2016 as part of the INFORM global surveillance program. PLoS One, 2018, 13(4)e0195281
[http://dx.doi.org/10.1371/journal.pone.0195281] [PMID: 29608599]
[35]
Luo, Q.; Yu, W.; Zhou, K.; Guo, L.; Shen, P.; Lu, H.; Huang, C.; Xu, H.; Xu, S.; Xiao, Y.; Li, L. Molecular epidemiology and colistin resistant mechanism of mcr-positive and mcr-negative clinical isolated Escherichia coli. Front. Microbiol., 2017, 8, 2262.
[http://dx.doi.org/10.3389/fmicb.2017.02262] [PMID: 29250039]

Rights & Permissions Print Cite
Article Metrics
29
1
© 2024 Bentham Science Publishers | Privacy Policy