1887
  1. Home
  2. Eurosurveillance
  3. Volume 23, Issue 6, 08/Feb/2018
  4. Article
Research article Open Access
Like 0
Download

Abstract

Background and aim

Plasmid-mediated colistin resistance mechanisms have been identified worldwide in the past years. A multiplex polymerase chain reaction (PCR) protocol for detection of all currently known transferable colistin resistance genes ( to , and variants) in was developed for surveillance or research purposes. We designed four new primer pairs to amplify , , and gene products and used the originally described primers for to obtain a stepwise separation of ca 200 bp between amplicons. The primer pairs and amplification conditions allow for single or multiple detection of all currently described genes and their variants present in . The protocol was validated testing 49 European and isolates of animal origin. Multiplex PCR results in bovine and porcine isolates from Spain, Germany, France and Italy showed full concordance with whole genome sequence data. The method was able to detect and as singletons or in different combinations as they were present in the test isolates. One new variant, , was also identified. This method allows rapid identification of -positive bacteria and overcomes the challenges of phenotypic detection of colistin resistance. The multiplex PCR should be particularly interesting in settings or laboratories with limited resources for performing genetic analysis as it provides information on the mechanism of colistin resistance without requiring genome sequencing.

© Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Closure
A corrected article has been published for this content:
Erratum for Euro Surveill. 2018;23(6)
Loading

Article metrics loading...

/content/10.2807/1560-7917.ES.2018.23.6.17-00672
2018-02-08
2024-12-22
/content/10.2807/1560-7917.ES.2018.23.6.17-00672
Loading
Loading full text...

Full text loading...

/deliver/fulltext/eurosurveillance/23/6/eurosurv-23-6-4.html?itemId=/content/10.2807/1560-7917.ES.2018.23.6.17-00672&mimeType=html&fmt=ahah

References

  1. Poirel L, Jayol A, Nordmann P. Polymyxins: Antibacterial activity, susceptibility testing, and resistance mechanisms encoded by plasmids or chromosomes. Clin Microbiol Rev. 2017;30(2):557-96.  https://doi.org/10.1128/CMR.00064-16  PMID: 28275006 
  2. Catry B, Cavaleri M, Baptiste K, Grave K, Grein K, Holm A, et al. Use of colistin-containing products within the European Union and European Economic Area (EU/EEA): development of resistance in animals and possible impact on human and animal health. Int J Antimicrob Agents. 2015;46(3):297-306.  https://doi.org/10.1016/j.ijantimicag.2015.06.005  PMID: 26215780 
  3. Rhouma M, Beaudry F, Thériault W, Letellier A. Colistin in Pig Production: Chemistry, Mechanism of Antibacterial Action, Microbial Resistance Emergence, and One Health Perspectives. Front Microbiol. 2016;7:1789. . https://doi.org/10.3389/fmicb.2016.01789 https://doi.org/10.3389/fmicb.2016.01789  PMID: 27891118 
  4. Liu YY, Wang Y, Walsh TR, Yi LX, Zhang R, Spencer J, et al. 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):161-8.  https://doi.org/10.1016/S1473-3099(15)00424-7  PMID: 26603172 
  5. European Medicines Agency (EMA). Updated advice on the use of colistin products in animals within the European Union: development of resistance and possible impact on human and animal health. London: EMA; Jul 2016. Available from: http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2016/07/WC500211080.pdf
  6. McPhee JB, Lewenza S, Hancock RE. Cationic antimicrobial peptides activate a two-component regulatory system, PmrA-PmrB, that regulates resistance to polymyxin B and cationic antimicrobial peptides in Pseudomonas aeruginosa. Mol Microbiol. 2003;50(1):205-17.  https://doi.org/10.1046/j.1365-2958.2003.03673.x  PMID: 14507375 
  7. European Committee on Antimicrobial Susceptibility Testing (EUCAST). EUCAST warnings concerning antimicrobial susceptibility testing products or procedures. Växjö: EUCAST; Jun 2017. Available from: http://www.eucast.org/ast_of_bacteria/warnings/
  8. European Committee on Antimicrobial Susceptibility Testing (EUCAST). Recommendations for MIC determination of colistin (polymyxin E). As recommended by the joint CLSI-EUCAST Polymyxin Breakpoints Working Group. Växjö: EUCAST; Jun 2016. Available from: http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/General_documents/Recommendations_for_MIC_determination_of_colistin_March_2016.pdf
  9. Clinical and Laboratory Standards Institute (CLSI) Subcommittee on Antimicrobial Susceptibility Testing. CLSI AST News Update Volume 1, Issue 2. Philadelphia: CLSI; Dec 2016. Available from: https://clsi.org/media/1700/clsi-news-winter-2016.pdf
  10. Shen Z, Wang Y, Shen Y, Shen J, Wu C. Early emergence of mcr-1 in Escherichia coli from food-producing animals. Lancet Infect Dis. 2016;16(3):293.  https://doi.org/10.1016/S1473-3099(16)00061-X  PMID: 26973308 
  11. Xavier BB, Lammens C, Ruhal R, Kumar-Singh S, Butaye P, Goossens H, et al. Identification of a novel plasmid-mediated colistin-resistance gene, mcr-2, in Escherichia coli, Belgium, June 2016. Euro Surveill. 2016;21(27):30280.  https://doi.org/10.2807/1560-7917.ES.2016.21.27.30280  PMID: 27416987 
  12. Yin W, Li H, Shen Y, Liu Z, Wang S, Shen Z, et al. Novel plasmid-mediated colistin resistance gene mcr-3 in Escherichia coli. MBio. 2017;8(3):e00543-17.  https://doi.org/10.1128/mBio.00543-17  PMID: 28655818 
  13. Carattoli A, Villa L, Feudi C, Curcio L, Orsini S, Luppi A, et al. 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.  https://doi.org/10.2807/1560-7917.ES.2017.22.31.30589  PMID: 28797329 
  14. Borowiak M, Fischer J, Hammerl JA, Hendriksen RS, 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):3317-24.  https://doi.org/10.1093/jac/dkx327  PMID: 28962028 
  15. Skov RL, Monnet DL. Plasmid-mediated colistin resistance (mcr-1 gene): three months later, the story unfolds. Euro Surveill. 2016;21(9):30155.  https://doi.org/10.2807/1560-7917.ES.2016.21.9.30155  PMID: 26967914 
  16. Lu X, Hu Y, Luo M, Zhou H, Wang X, Du Y, et al. MCR-1.6, a New MCR variant carried by an IncP plasmid in a colistin-resistant Salmonella enterica serovar Typhimurium isolate from a healthy individual. Antimicrob Agents Chemother. 2017;61(5):e02632-16.  https://doi.org/10.1128/AAC.02632-16  PMID: 28264851 
  17. Roer L, Hansen F, Stegger M, Sönksen UW, Hasman H, Hammerum AM. Novel mcr-3 variant, encoding mobile colistin resistance, in an ST131 Escherichia coli isolate from bloodstream infection, Denmark, 2014. Euro Surveill. 2017;22(31):30584.  https://doi.org/10.2807/1560-7917.ES.2017.22.31.30584  PMID: 28797324 
  18. Carretto E, Brovarone F, Nardini P, Russello G, Barbarini D, Pongolini S, et al. Detection of mcr-4 positive Salmonella enterica serovar Typhimurium in clinical isolates of human origin, Italy, October to November 2016. Euro Surveill. 2018. 23(2):pii=17-00821. https://doi.org/10.2807/1560-7917.ES.2018.23.2.17-00821
  19. Litrup E, Kiil K, Hammerum AM, Roer L, Nielsen EM, Torpdahl M. Plasmid-borne colistin resistance gene mcr-3 in Salmonella isolates from human infections, Denmark, 2009-17. Euro Surveill. 2017;22(31):30587.  https://doi.org/10.2807/1560-7917.ES.2017.22.31.30587  PMID: 28797325 
  20. Hernández M, Iglesias MR, Rodríguez-Lázaro D, Gallardo A, Quijada N, Miguela-Villoldo P, et al. Co-occurrence of colistin-resistance genes mcr-1 and mcr-3 among multidrug-resistant Escherichia coli isolated from cattle, Spain, September 2015. Euro Surveill. 2017;22(31):30586.  https://doi.org/10.2807/1560-7917.ES.2017.22.31.30586  PMID: 28797328 
  21. Kluytmans J. Plasmid-encoded colistin resistance: mcr-one, two, three and counting. Euro Surveill. 2017;22(31):30588.  https://doi.org/10.2807/1560-7917.ES.2017.22.31.30588  PMID: 28797321 
  22. Hasman H, Hammerum AM, Hansen F, Hendriksen RS, Olesen B, Agersø Y, et al. Detection of mcr-1 encoding plasmid-mediated colistin-resistant Escherichia coli isolates from human bloodstream infection and imported chicken meat, Denmark 2015. Euro Surveill. 2015;20(49):30085.  https://doi.org/10.2807/1560-7917.ES.2015.20.49.30085  PMID: 26676364 
  23. Di Pilato V, Arena F, Tascini C, Cannatelli A, Henrici De Angelis L, Fortunato S, et al. mcr-1.2, a new mcr variant carried on a transferable plasmid from a colistin-resistant KPC carbapenemase-producing Klebsiella pneumoniae strain of Sequence Type 512. Antimicrob Agents Chemother. 2016;60(9):5612-5.  https://doi.org/10.1128/AAC.01075-16  PMID: 27401575 
  24. von Wintersdorff CJ, Wolffs PF, van Niekerk JM, Beuken E, van Alphen LB, Stobberingh EE, et al. Detection of the plasmid-mediated colistin-resistance gene mcr-1 in faecal metagenomes of Dutch travellers. J Antimicrob Chemother. 2016;71(12):3416-9.  https://doi.org/10.1093/jac/dkw328  PMID: 27559117 
  25. Leangapichart T, Gautret P, Brouqui P, Mimish Z, Raoult D, Rolain J-M. Acquisition of mcr-1 plasmid-mediated colistin resistance in Escherichia coli and Klebsiella pneumoniae during Hajj 2013 and 2014. Antimicrob Agents Chemother. 2016;60(11):6998-9.  https://doi.org/10.1128/AAC.01486-16  PMID: 27620480 
  26. European Commission. Commission implementing decision 2013/652/EU on the monitoring and reporting of antimicrobial resistance in zoonotic and commensal bacteria. Official Journal of the European Union. Luxembourg: Publications Office of the European Union. 14.11.2013:L 303. Available from: http://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri=OJ:L:2013:303:FULL&from=EN
  27. Brennan E, Martins M, McCusker MP, Wang J, Alves BM, Hurley D, et al. Multidrug-Resistant Escherichia coli in Bovine Animals, Europe. Emerg Infect Dis. 2016;22(9):1650-2.  https://doi.org/10.3201/eid2209.160140  PMID: 27533105 
  28. Quesada A, Porrero MC, Téllez S, Palomo G, García M, Domínguez L. Polymorphism of genes encoding PmrAB in colistin-resistant strains of Escherichia coli and Salmonella enterica isolated from poultry and swine. J Antimicrob Chemother. 2015;70(1):71-4.  https://doi.org/10.1093/jac/dku320  PMID: 25150146 
  29. The European Committee on Antimicrobial Susceptibility Testing (EUCAST). Breakpoint tables for interpretation of MICs and zone diameters. Version 8.0. Växjö: EUCAST; 2018. Available from: http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Breakpoint_tables/v_8.0_Breakpoint_Tables.pdf
Multiplex PCR for detection of plasmid-mediated colistin resistance determinants, mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5 for surveillance purposes
<p class="citation"> <span>Citation style for this article:</span> <span class="meta-value authors"> <a href="/search?value1=Ana+Rita+Rebelo&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Rebelo Ana Rita</a>, <a href="/search?value1=Valeria+Bortolaia&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Bortolaia Valeria</a>, <a href="/search?value1=Jette+S+Kjeldgaard&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Kjeldgaard Jette S</a>, <a href="/search?value1=Susanne+K+Pedersen&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Pedersen Susanne K</a>, <a href="/search?value1=Pimlapas+Leekitcharoenphon&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Leekitcharoenphon Pimlapas</a>, <a href="/search?value1=Inge+M+Hansen&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Hansen Inge M</a>, <a href="/search?value1=Beatriz+Guerra&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Guerra Beatriz</a>, <a href="/search?value1=Burkhard+Malorny&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Malorny Burkhard</a>, <a href="/search?value1=Maria+Borowiak&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Borowiak Maria</a>, <a href="/search?value1=Jens+Andre+Hammerl&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Hammerl Jens Andre</a>, <a href="/search?value1=Antonio+Battisti&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Battisti Antonio</a>, <a href="/search?value1=Alessia+Franco&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Franco Alessia</a>, <a href="/search?value1=Patricia+Alba&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Alba Patricia</a>, <a href="/search?value1=Agnes+Perrin-Guyomard&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Perrin-Guyomard Agnes</a>, <a href="/search?value1=Sophie+A+Granier&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Granier Sophie A</a>, <a href="/search?value1=Cristina++De+Frutos+Escobar&option1=author&noRedirect=true" class="nonDisambigAuthorLink">De Frutos Escobar Cristina </a>, <a href="/search?value1=Surbhi+Malhotra-Kumar&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Malhotra-Kumar Surbhi</a>, <a href="/search?value1=Laura+Villa&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Villa Laura</a>, <a href="/search?value1=Alessandra+Carattoli&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Carattoli Alessandra</a>, <a href="/search?value1=Rene+S+Hendriksen&option1=author&noRedirect=true" class="nonDisambigAuthorLink">Hendriksen Rene S</a></span>. Multiplex PCR for detection of plasmid-mediated colistin resistance determinants, mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5 for surveillance purposes. <a href="/content/ecdc">Euro Surveill.</a> 2018;23(6):pii=17-00672. <a href="https://doi.org/10.2807/1560-7917.ES.2018.23.6.17-00672" title="doi link" target="_blank">https://doi.org/10.2807/1560-7917.ES.2018.23.6.17-00672</a> <span class="ES_Article_citation"> <span class="generated">Received</span>: 04 Oct 2017;&nbsp;&nbsp; <span class="generated">Accepted</span>: 13 Nov 2017 </span> </p>
/content/10.2807/1560-7917.ES.2018.23.6.17-00672
/content/10.2807/1560-7917.ES.2018.23.6.17-00672
Loading

Data & Media loading...

Supplementary data

Submit comment
Close
Comment moderation successfully completed
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error