Pulsed-Field Gel Electrophoresis (PFGE): Application in Population Structure Studies of Bovine Mastitis-Causing Streptococci

  • Ilda Santos-Sanches
  • Lélia Chambel
  • Rogério Tenreiro
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1247)

Abstract

Pulsed-field gel electrophoresis (PFGE) separates large DNA molecules by the use of an alternating electrical field, such that greater size resolution can be obtained when compared to normal agarose gel electrophoresis. PFGE is often employed to track pathogens and is a valuable typing scheme to detect and differentiate strains. Particularly, the contour-clamped homogeneous electric field (CHEF) PFGE system is considered to be the gold standard for use in epidemiological studies of many bacterial pathogens. Here we describe a PFGE protocol that was applicable to the study of bovine streptococci, namely, Streptococcus agalactiae (group B Streptococcus, GBS), Streptococcus dysgalactiae subsp. dysgalactiae (group C Streptococcus, GCS), and Streptococcus uberis—which are relevant pathogens causing mastitis, a highly prevalent and costly disease in dairy industry due to antibiotherapy and loss in milk production.

Key words

Bovine mastitis Streptococcus agalactiae Streptococcus dysgalactiae subsp. dysgalactiae Streptococcus uberis Molecular typing Molecular epidemiology Pulsed-field gel electrophoresis PFGE Alternating electrical field CHEF 

References

  1. 1.
    Tenover FC, Arbeit RD, Goering RV et al (1995) Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol 33:2233–2239PubMedCentralPubMedGoogle Scholar
  2. 2.
    Van Belkum A, Tassios PT, Dijkshoorn L et al (2007) Guidelines for the validation and application of typing methods for the use in bacterial epidemiology. Clin Microbiol Infect 13(suppl 3):1–46PubMedCrossRefGoogle Scholar
  3. 3.
    Ericsson U, Lindberg A, Persson W et al (2009) Microbial aetiology of acute clinical mastitis and agent-specific risk factors. Vet Microbiol 137:90–97CrossRefGoogle Scholar
  4. 4.
    Barkema HW, Green MJ, Bradley AJ, Zadoks RN (2009) Invited review: the role of contagious disease in udder health. J Dairy Sci 92:4717–4729PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Muellner P, Zadoks RN, Perez AM et al (2011) The integration of molecular tools into veterinary and spatial epidemiology. Spat Spattemporal Epidemiol 2:159–171CrossRefGoogle Scholar
  6. 6.
    Rato MG, Bexiga R, Nunes SF et al (2008) Molecular epidemiology and population structure of bovine Streptococcus uberis. J Dairy Sci 91:4542–4551PubMedCrossRefGoogle Scholar
  7. 7.
    Rato MG, Bexiga R, Nunes SF, Vilela CL, Santos-Sanches I (2010) Human group A streptococci virulence genes in bovine group C streptococci. Emerg Infect Dis 16:116–119PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Rato MG, Nerlich A, Bergmann R et al (2011) Virulence gene pool detected in bovine group C Streptococcus dysgalactiae subsp. dysgalactiae using a group A Streptococcus pyogenes virulence microarray. J Clin Microbiol 49:2470–2479PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Rato MG, Bexiga R, Florindo C et al (2013) Antimicrobial resistance and molecular epidemiology of streptococci from bovine mastitis. Vet Microbiol 161:286–294PubMedCrossRefGoogle Scholar
  10. 10.
    Rato M (2011) Epidemiological characterization, antimicrobial resistance and virulence mechanisms in animal and human streptococci. Ph.D. thesis, Faculdade de Ciências e Tecnologia. Universidade Nova de Lisboa. Portugal. ISBN: 978-989-20-2618-3Google Scholar
  11. 11.
    Chung M, de Lencastre H, Matthews P et al (2000) Molecular typing of methicillin-resistant Staphylococcus aureus by pulsed-field gel electrophoresis: comparison of results obtained in a multilaboratory effort using identical protocols and MRSA strains. Microb Drug Resist 6:189–198PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Ilda Santos-Sanches
    • 1
  • Lélia Chambel
    • 2
  • Rogério Tenreiro
    • 2
  1. 1.Faculdade de Ciências e Tecnologia (FCT), Centro de Recursos Microbiológicos (CREM) and Research Unit on Applied Molecular Biosciences (UCIBIO, REQUIMTE)Universidade Nova de Lisboa (UNL)Caparica, LisbonPortugal
  2. 2.Faculdade de Ciências (FC), Centro de Biodiversidade, Genómica Integrativa e Funcional (BioFig)Universidade de Lisboa (UL)LisbonPortugal

Personalised recommendations