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Veterinary Infection Biology: Molecular Diagnostics and High-Throughput Strategies

Volume 1247 of the series Methods in Molecular Biology pp 323-334

Date:

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

  • Ilda Santos-SanchesAffiliated withFaculdade de Ciências e Tecnologia (FCT), Centro de Recursos Microbiológicos (CREM) and Research Unit on Applied Molecular Biosciences (UCIBIO, REQUIMTE), Universidade Nova de Lisboa (UNL) Email author 
  • , Lélia ChambelAffiliated withFaculdade de Ciências (FC), Centro de Biodiversidade, Genómica Integrativa e Funcional (BioFig), Universidade de Lisboa (UL)
  • , Rogério TenreiroAffiliated withFaculdade de Ciências (FC), Centro de Biodiversidade, Genómica Integrativa e Funcional (BioFig), Universidade de Lisboa (UL)

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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