title: Discrimination between Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capricolum using PCR-RFLP and PCR.
abstract: In this study, the dihydrolipoyl dehydrogenase (lpdA) gene was used to distinguish Mycoplasma mycoides subsp. capri (Mmc) from Mycoplasma capricolum subsp. capricolum (Mcc), two of four Mycoplasma species that cause contagious agalactia in sheep and goats. After alignment of nucleotide sequences of both species, specific primer sets were designed from unchanging and variable gene segments. The first primer set LPD-C1-F/LPD-C1-R was used to amplify a 911 bp fragment that was subsequently co-digested with FastDigest PstI, SspI, EcoRI and ClaI enzymes. The PCR-RFLP profiles differentiated the two mycoplasma species. The second primer set was used to distinguish Mmc from Mcc by single tube PCR. Both methods were further applied to identify 54 isolates collected from dairy herds from different provinces in Sardinia. The results of this study showed that PCR-RFLP and PCR could be used in routine diagnosis for rapid and specific simultaneous discrimination of Mmc and Mcc.
authors.affiliation: Istituto Zooprofilattico Sperimentale della Sardegna G. Pegreffi, 07100 Sassari, Italy.
authors.forename: Grazia
authors.initials: G
authors.lastname: Cillara
authors.affiliation: Istituto Zooprofilattico Sperimentale della Sardegna G. Pegreffi, 07100 Sassari, Italy.
authors.forename: Maria Giovanna
authors.initials: MG
authors.lastname: Manca
authors.affiliation: Istituto Zooprofilattico Sperimentale della Sardegna G. Pegreffi, 07100 Sassari, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Longheu
authors.affiliation: Istituto Zooprofilattico Sperimentale della Sardegna G. Pegreffi, 07100 Sassari, Italy. Electronic address: sebastiana.tola@izs-sardegna.it.
authors.forename: Sebastiana
authors.initials: S
authors.lastname: Tola
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology; D005798:Genes, Bacterial; D009174:Mycoplasma / Q000145:classification / Q000235:genetics; D009175:Mycoplasma Infections / Q000382:microbiology / Q000662:veterinary; D045623:Mycoplasma capricolum / Q000145:classification* / Q000235:genetics; D009176:Mycoplasma mycoides / Q000145:classification* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D012150:Polymorphism, Restriction Fragment Length; D013045:Species Specificity
keywords: Contagious agalactia; Dihydrolipoyl dehydrogenase; Mycoplasma capricolum subsp. capricolum; Mycoplasma mycoides subsp. capri; PCR; PCR-RFLP