title: Rapid differential diagnosis of Mycoplasma agalactiae and Mycoplasma bovis based on a multiplex-PCR and a PCR-RFLP.
abstract: The membrane-protein 81 gene (mb-mp81) of Mycoplasma bovis was cloned, sequenced and compared to membrane-protein 81 gene (ma-mp81) of Mycoplasma agalactiae. After alignment of both sequences, specific primers pairs were designed from variable or unchanging nucleotide segments. In this study, we describe the development and optimization of a multiplex-PCR (MPCR) for the rapid detection of M. agalactiae and M. bovis strains. In addition, a simple and rapid PCR-restriction fragment length polymorphism (RFLP) assay, using the restriction enzymes AluI, DraI, RsaI and XbaI, is described to distinguish between both species. The results suggest that MPCR and PCR-RFLP assays could be used as an alternative method in routine diagnosis for rapid and specific simultaneous detection of M. agalactiae and M. bovis.
authors.affiliation: Istituto Zooprofilattico Sperimentale della Sardegna G.Pegreffi, Via Duca degli Abruzzi 8, 07100 Sassari, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Foddai
authors.forename: G
authors.initials: G
authors.lastname: Idini
authors.forename: M
authors.initials: M
authors.lastname: Fusco
authors.forename: N
authors.initials: N
authors.lastname: Rosa
authors.forename: C
authors.initials: C
authors.lastname: de la Fe
authors.forename: S
authors.initials: S
authors.lastname: Zinellu
authors.forename: L
authors.initials: L
authors.lastname: Corona
authors.forename: S
authors.initials: S
authors.lastname: Tola
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D003937:Diagnosis, Differential; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics; D008970:Molecular Weight; D045644:Mycoplasma agalactiae / Q000235:genetics* / Q000302:isolation & purification*; D045682:Mycoplasma bovis / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins