Pulsed-field gel electrophoresis (PFGE): application in population structure studies of bovine mastitis-causing streptococci.
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.
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), Campus de Caparica, 2829-516, Caparica, Lisbon, Portugal, imgs@fct.unl.pt.
Ilda
I
Santos-Sanches
Lélia
L
Chambel
Rogério
R
Tenreiro
D000818:Animals; D002417:Cattle; D016172:DNA Fingerprinting; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D005260:Female; D008414:Mastitis, Bovine / Q000175:diagnosis* / Q000382:microbiology*; D058889:Molecular Typing; D013290:Streptococcal Infections / Q000662:veterinary*; D013291:Streptococcus / Q000235:genetics*