Skip to main content

PLASmid TAXonomic PCR (PlasTax-PCR), a Multiplex Relaxase MOB Typing to Assort Plasmids into Taxonomic Units

Part of the Methods in Molecular Biology book series (MIMB,volume 2392)


Plasmids transmissible by conjugation are responsible for disseminating antibiotic-resistance genes, making plasmid detection relevant for pathogen tracking. We describe the use of a multiplex PCR method for the experimental identification of specific plasmid taxonomic units (PTUs) of transmissible plasmids. The PCR primers were designed to target conserved segments of the relaxase MOB gene of PTUs encoding adaptive traits for enterobacteria (antimicrobial resistance, virulence, and metabolism). In this way, PlasTax-PCR detects the presence of these plasmids and allows their direct assignation to a PTU.

Key words

  • Plasmid taxonomic units
  • Bacterial conjugation
  • Horizontal gene transfer
  • Relaxase
  • MOB family
  • Plasmid typing

This is a preview of subscription content, access via your institution.

Buying options

USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1007/978-1-0716-1799-1_10
  • Chapter length: 16 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
USD   109.00
Price excludes VAT (USA)
  • ISBN: 978-1-0716-1799-1
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   149.99
Price excludes VAT (USA)
Hardcover Book
USD   219.99
Price excludes VAT (USA)
Fig. 1
Fig. 2

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more


  1. de la Cruz F, Davies J (2000) Horizontal gene transfer and the origin of species: lessons from bacteria. Trends Microbiol 8(3):128–133.

    CrossRef  Google Scholar 

  2. Garcillán-Barcia MP, Francia MV, de la Cruz F (2009) The diversity of conjugative relaxases and its application in plasmid classification. FEMS Microbiol Rev 33(3):657–687.

    CrossRef  Google Scholar 

  3. Garcillán-Barcia MP, Redondo-Salvo S, Vielva L, de la Cruz F (2020) MOBscan: automated annotation of MOB relaxases. In: de la Cruz F (ed) Horizontal gene transfer: methods and protocols, Methods in molecular biology. Science+Business Media, LLC, part of Springer Nature, New York, NY, pp 295–308

    CrossRef  Google Scholar 

  4. Redondo-Salvo S, Fernández-López R, Ruiz R, Vielva L, de Toro M, Rocha EPC et al (2020) Pathways for horizontal gene transfer in bacteria revealed by a global map of their plasmids. Nat Commun 11(1):3602.

    CrossRef  Google Scholar 

  5. Garcillán-Barcia MP, Alvarado A, de la Cruz F (2011) Identification of bacterial plasmids based on mobility and plasmid population biology. FEMS Microbiol Rev 35(5):936–956.

    CrossRef  Google Scholar 

  6. Alvarado A, Garcillán-Barcia MP, de la Cruz F (2012) A degenerate primer MOB typing (DPMT) method to classify gamma-proteobacterial plasmids in clinical and environmental settings. PLoS One 7(7):e40438.

    CrossRef  CAS  Google Scholar 

  7. Villa L, Carattoli A (2020) Plasmid typing and classification. Springer, New York, pp 309–321.

    CrossRef  Google Scholar 

  8. Valverde A, Cantón R, Garcillán-Barcia MP, Novais A, Galán JC, Alvarado A et al (2009) Spread of bla(CTX-M-14) is driven mainly by IncK plasmids disseminated among Escherichia coli phylogroups A, B1, and D in Spain. Antimicrob Agents Chemother 53(12):5204–5212.

    CrossRef  CAS  Google Scholar 

  9. Mata C, Miró E, Alvarado A, Garcillán-Barcia MP, Toleman M, Walsh TR et al (2012) Plasmid typing and genetic context of AmpC β-lactamases in Enterobacteriaceae lacking inducible chromosomal ampC genes: findings from a Spanish hospital 1999-2007. J Antimicrob Chemother 67(1):115–122.

    CrossRef  CAS  Google Scholar 

  10. Coelho A, Piedra-Carrasco N, Bartolomé R, Quintero-Zarate JN, Larrosa N, Cornejo-Sánchez T et al (2012) Role of IncHI2 plasmids harbouring blaVIM-1, blaCTX-M-9, aac(6’)-Ib and qnrA genes in the spread of multiresistant Enterobacter cloacae and Klebsiella pneumoniae strains in different units at Hospital Vall d’Hebron, Barcelona, Spain. Int J Antimicrob Agents 39(6):514–517.

    CrossRef  CAS  Google Scholar 

  11. Garcillán-Barcia MP, Ruiz del Castillo B, Alvarado A, de la Cruz F, Martínez-Martínez L (2015) Degenerate primer MOB typing of multiresistant clinical isolates of E. coli uncovers new plasmid backbones. Plasmid 77:17–27.

    CrossRef  Google Scholar 

  12. Adelowo OO, Caucci S, Banjo OA, Nnanna OC, Awotipe EO, Peters FB et al (2018) Extended Spectrum Beta-lactamase (ESBL)-producing bacteria isolated from hospital wastewaters, rivers and aquaculture sources in Nigeria. Environ Sci Pollut Res 25(3):2744–2755.

    CrossRef  CAS  Google Scholar 

  13. Turner S, Pryer KM, Miao VPW, Palmer JD (1999) Investigating deep phylogenetic relationships among cyanobacteria and plastids by small subunit rRNA sequence analysis. J Eukaryot Microbiol 46(4):327–338.

    CrossRef  CAS  PubMed  Google Scholar 

  14. Lanza VF, de Toro M, Garcillán-Barcia MP, Mora A, Blanco J, Coque TM et al (2014) Plasmid flux in Escherichia coli ST131 sublineages, analyzed by plasmid constellation network (PLACNET), a new method for plasmid reconstruction from whole genome sequences. PLoS Genet 10(12):e1004766

    CrossRef  Google Scholar 

  15. Coque TM, Novais Â, Carattoli A, Poirel L, Pitout J, Peixe L et al (2008) Dissemination of clonally related Escherichia coli strains expressing extended-Spectrum β-lactamase CTX-M-15. Emerg Infect Dis 14(2):195–200.

    CrossRef  CAS  Google Scholar 

  16. Novais A, Pires J, Ferreira H, Costa L, Montenegro C, Vuotto C et al (2012) Characterization of globally spread Escherichia coli ST131 isolates (1991 to 2010). Antimicrob Agents Chemother 56(7):3973–3976.

    CrossRef  CAS  Google Scholar 

Download references


This work was financed by the Spanish Ministry of Science and Innovation (PID2020-117923GB-I00) to FdlC and MPG-B, and by the European Commission, the Joint Programming Initiative in Antimicrobial Resistance and the Ministry of Science and Innovation (UE/STARCS AC16/00039, UE/ST131TS AC16/00043, and PI18/01942) to TMC.

Author information

Authors and Affiliations


Corresponding author

Correspondence to M. Pilar Garcillán-Barcia .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

© 2022 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Verify currency and authenticity via CrossMark

Cite this protocol

Cuartas, R., Coque, T.M., de la Cruz, F., Garcillán-Barcia, M.P. (2022). PLASmid TAXonomic PCR (PlasTax-PCR), a Multiplex Relaxase MOB Typing to Assort Plasmids into Taxonomic Units. In: Basu, C. (eds) PCR Primer Design. Methods in Molecular Biology, vol 2392. Humana, New York, NY.

Download citation

  • DOI:

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1798-4

  • Online ISBN: 978-1-0716-1799-1

  • eBook Packages: Springer Protocols