title: Development of a multiplex PCR assay for polymorphism analysis of Brucella suis biovars causing brucellosis in swine.
abstract: Swine brucellosis is caused by the biovars 1, 2 and 3 of Brucella suis the identification of which up to now relies on microbiological tests lacking adequate specificity together with time consuming and expensive molecular procedures. Based on sequence variation of the omp2b gene, we have developed a four primer set multiplex PCR assay that was tested for polymorphism analysis of B. suis biovars causing brucellosis in swine. The assay exploits the single nucleotide polymorphisms found in omp2b gene of B. suis reference biovars which are conserved in 43 B. suis field isolates from different geographic origins and hosts. Three specific amplification patterns (S1, S2 and S3) were obtained for reference strains of B. suis biovars 1, 2 and 3, respectively. However, some B. suis field isolates identified as biovars 2 or 3 according AMOS-PCR, PCR-RFLP of omp31 and omp2 genes and classical bacteriological methods, resulted also in S1 patterns, limiting the typing usefulness of the method.
authors.affiliation: Laboratório Nacional de Investigação Veterinária, Porto, Portugal. lenia.beck@lniv.min-agricultura.pt
authors.forename: Lénia
authors.initials: L
authors.lastname: Ferrão-Beck
authors.forename: Regina
authors.initials: R
authors.lastname: Cardoso
authors.forename: Pilar María
authors.initials: PM
authors.lastname: Muñoz
authors.forename: María Jesús
authors.initials: MJ
authors.lastname: de Miguel
authors.forename: David
authors.initials: D
authors.lastname: Albert
authors.forename: Ana Cristina
authors.initials: AC
authors.lastname: Ferreira
authors.forename: Clara María
authors.initials: CM
authors.lastname: Marín
authors.forename: Martine
authors.initials: M
authors.lastname: Thiébaud
authors.forename: Isabelle
authors.initials: I
authors.lastname: Jacques
authors.forename: Maggy
authors.initials: M
authors.lastname: Grayon
authors.forename: Michel S
authors.initials: MS
authors.lastname: Zygmunt
authors.forename: Bruno
authors.initials: B
authors.lastname: Garin-Bastuji
authors.forename: Jose María
authors.initials: JM
authors.lastname: Blasco
authors.forename: Maria Inácia
authors.initials: MI
authors.lastname: Sá
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D041021:Brucella suis / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D002006:Brucellosis / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D004269:DNA, Bacterial / Q000737:chemistry; D005784:Gene Amplification; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011110:Polymorphism, Genetic; D020641:Polymorphism, Single Nucleotide; D018272:Porins / Q000737:chemistry / Q000235:genetics*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000382:microbiology*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; C096736:Omp2b protein, Brucella; D018272:Porins