title: Molecular identification of Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum by a duplex PCR assay.
abstract: Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum (S Gallinarum) and biovar Pullorum (S Pullorum) are 2 poultry pathogens that cause major economic losses to the poultry industry worldwide. Control of both diseases mainly relies on the adoption of biosecurity programs, and success is dependent on accurate and fast detection. Based on this concept, we developed a duplex PCR assay, targeting 2 chromosomal sequences, which allowed us to precisely identify and differentiate S Gallinarum and S Pullorum field strains. This assay was validated by testing genomic DNA from 40 S Gallinarum and 29 S Pullorum field strains, 87 other Salmonella serovars, and 7 non-Salmonella strains. The serovar identifier region (SIR) primers produced a fragment only in S Gallinarum and S Pullorum strains, whereas the fragment from the ratA coding sequence, which was previously demonstrated to differentiate the 2 biovars, was also amplified from other Salmonella serovars. Our results showed that the combination of both SIR and ratA amplifications could be used to identify as well as to differentiate colonies of S Gallinarum and S Pullorum reliably. Thus, we believe this methodology can be a useful ancillary tool for routine veterinary diagnostic laboratories by providing rapid, accurate results.
authors.affiliation: Faculty of Agriculture and Veterinary Sciences, São Paulo State University, Jaboticabal, São Paulo, Brazil (Batista, Almeida, Barbosa, Berchieri Jr)Agronomical Science Centre, Federal University of Paraíba, Campus II, Areia, Paraíba, Brazil (Freitas Neto)School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington, Leicestershire, United Kingdom (Barrow).
authors.forename: Diego Felipe Alves
authors.initials: DF
authors.lastname: Batista
authors.affiliation: Faculty of Agriculture and Veterinary Sciences, São Paulo State University, Jaboticabal, São Paulo, Brazil (Batista, Almeida, Barbosa, Berchieri Jr)Agronomical Science Centre, Federal University of Paraíba, Campus II, Areia, Paraíba, Brazil (Freitas Neto)School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington, Leicestershire, United Kingdom (Barrow) oliveirocaetano@yahoo.com.br.
authors.forename: Oliveiro Caetano
authors.initials: OC
authors.lastname: de Freitas Neto
authors.affiliation: Faculty of Agriculture and Veterinary Sciences, São Paulo State University, Jaboticabal, São Paulo, Brazil (Batista, Almeida, Barbosa, Berchieri Jr)Agronomical Science Centre, Federal University of Paraíba, Campus II, Areia, Paraíba, Brazil (Freitas Neto)School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington, Leicestershire, United Kingdom (Barrow).
authors.forename: Adriana Maria
authors.initials: AM
authors.lastname: de Almeida
authors.affiliation: Faculty of Agriculture and Veterinary Sciences, São Paulo State University, Jaboticabal, São Paulo, Brazil (Batista, Almeida, Barbosa, Berchieri Jr)Agronomical Science Centre, Federal University of Paraíba, Campus II, Areia, Paraíba, Brazil (Freitas Neto)School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington, Leicestershire, United Kingdom (Barrow).
authors.forename: Paul Andrew
authors.initials: PA
authors.lastname: Barrow
authors.affiliation: Faculty of Agriculture and Veterinary Sciences, São Paulo State University, Jaboticabal, São Paulo, Brazil (Batista, Almeida, Barbosa, Berchieri Jr)Agronomical Science Centre, Federal University of Paraíba, Campus II, Areia, Paraíba, Brazil (Freitas Neto)School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington, Leicestershire, United Kingdom (Barrow).
authors.forename: Fernanda
authors.initials: F
authors.lastname: de Oliveira Barbosa
authors.affiliation: Faculty of Agriculture and Veterinary Sciences, São Paulo State University, Jaboticabal, São Paulo, Brazil (Batista, Almeida, Barbosa, Berchieri Jr)Agronomical Science Centre, Federal University of Paraíba, Campus II, Areia, Paraíba, Brazil (Freitas Neto)School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington, Leicestershire, United Kingdom (Barrow).
authors.forename: Angelo
authors.initials: A
authors.lastname: Berchieri Junior
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D002645:Chickens; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000382:microbiology; D012481:Salmonella Infections, Animal / Q000175:diagnosis* / Q000382:microbiology; D019779:Salmonella enterica / Q000145:classification* / Q000235:genetics; D065288:Serogroup
chemical_list: D001426:Bacterial Proteins
keywords: Duplex PCR; Salmonella Gallinarum; Salmonella Pullorum; fowl typhoid; pullorum disease