title: Genetic evaluation of IS900 partial sequence of Mycobacterium avium subsp. paratuberculosis Brazilian isolates from bovine milk.
abstract: Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of paratuberculosis. Insertion sequence IS900 is used for the identification of MAP. The objective of this study was to verify the genetic conservation of IS900 sequences in raw milk samples. To evaluate genetic conservation, 206 quarter milk samples and 16 bulk-tank milk samples were collected. DNA extraction and IS900 PCR were performed in all samples. Six samples amplified the expected fragment. To confirm the identity of the amplified fragments, PCR products were cloned and sequenced. The resulting sequences were compared with other MAP sequences from GenBank, and it was possible to identify eight polymorphic regions and to form five distinct haplotypes. The number of mutations in each haplotype was verified. IS900 sequence is a very well-conserved sequence that could be used as tool for the molecular detection of this agent and epidemiological purposes. The results showed the first genetic analysis on Brazilian isolates of MAP.
authors.affiliation: Setor de Medicina Veterinária Preventiva e Saúde Pública, Departamento de Veterinária, Universidade Federal de Viçosa, Minas Gerais, 36570-000, Brazil.
authors.forename: Isabel Azevedo
authors.initials: IA
authors.lastname: Carvalho
authors.forename: Victor Oliveira
authors.initials: VO
authors.lastname: Silva
authors.forename: Pedro Marcus Pereira
authors.initials: PM
authors.lastname: Vidigal
authors.forename: Abelardo
authors.initials: A
authors.lastname: Silva
authors.forename: Maria Aparecida Scatamburlo
authors.initials: MA
authors.lastname: Moreira
mesh_terms: D000818:Animals; D001938:Brazil / Q000453:epidemiology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000453:epidemiology* / Q000635:transmission; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics*; D006239:Haplotypes; D008892:Milk / Q000737:chemistry / Q000382:microbiology*; D008969:Molecular Sequence Data; D016927:Mycobacterium avium subsp. paratuberculosis / Q000235:genetics* / Q000302:isolation & purification; D010283:Paratuberculosis / Q000175:diagnosis / Q000453:epidemiology* / Q000635:transmission; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011110:Polymorphism, Genetic; D016415:Sequence Alignment / Q000662:veterinary; D017422:Sequence Analysis, DNA / Q000662:veterinary
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial