Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Feb;23(6):17-00672.
doi: 10.2807/1560-7917.ES.2018.23.6.17-00672.

Multiplex PCR for detection of plasmid-mediated colistin resistance determinants, mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5 for surveillance purposes

Ana Rita Rebelo  1 , Valeria Bortolaia  1 , Jette S Kjeldgaard  1 , Susanne K Pedersen  1 , Pimlapas Leekitcharoenphon  1 , Inge M Hansen  1 , Beatriz Guerra  2 , Burkhard Malorny  3 , Maria Borowiak  3 , Jens Andre Hammerl  3 , Antonio Battisti  4 , Alessia Franco  4 , Patricia Alba  4 , Agnes Perrin-Guyomard  5 , Sophie A Granier  6 , Cristina De Frutos Escobar  7 , Surbhi Malhotra-Kumar  8 , Laura Villa  9 , Alessandra Carattoli  9 , Rene S Hendriksen  1
Affiliations

Multiplex PCR for detection of plasmid-mediated colistin resistance determinants, mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5 for surveillance purposes

Ana Rita Rebelo et al. Euro Surveill. 2018 Feb.

Erratum in

  • Erratum for Euro Surveill. 2018;23(6).
    Eurosurveillance editorial team. Eurosurveillance editorial team. Euro Surveill. 2018 Feb;23(7):180215-1. doi: 10.2807/1560-7917.ES.2018.23.7.180215-1. Euro Surveill. 2018. PMID: 29471625 Free PMC article. No abstract available.

Abstract

Background and aimPlasmid-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 (mcr-1 to mcr-5, and variants) in Enterobacteriaceae was developed for surveillance or research purposes. Methods: We designed four new primer pairs to amplify mcr-1, mcr-2, mcr-3 and mcr-4 gene products and used the originally described primers for mcr-5 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 mcr genes and their variants present in Enterobacteriaceae. The protocol was validated testing 49 European Escherichia coli and Salmonella isolates of animal origin. Results: 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 mcr-1, mcr-3 and mcr-4 as singletons or in different combinations as they were present in the test isolates. One new mcr-4 variant, mcr-4.3, was also identified. Conclusions: This method allows rapid identification of mcr-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.

Keywords: antimicrobial resistance; colistin; mcr; mcr-4.3; multiplex; polymerase chain reaction; polymyxin E; surveillance; transferable resistance.

PubMed Disclaimer

Conflict of interest statement

Conflict of interest: None declared.

Figures

Figure
Figure
Multiplex PCR for detection of mcr-1, mcr-2, mcr-3, mcr-4 and mcr-5, European Union Reference Laboratory for Antimicrobial Resistance (EURL-AR) in the context of animal health and food safety, 2017
See this image and copyright information in PMC

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. 10.1128/CMR.00064-16 - DOI - PMC - PubMed
    1. 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. 10.1016/j.ijantimicag.2015.06.005 - DOI - PubMed
    1. 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 10.3389/fmicb.2016.01789 - DOI - DOI - PMC - PubMed
    1. 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. 10.1016/S1473-3099(15)00424-7 - DOI - PubMed
    1. 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_guidelin...

MeSH terms