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In situ-PCR for the detection of Mycobacterium paratuberculosis DNA in paraffin-embedded tissues.
Sanna, E; Woodall, C J; Watt, N J; Clarke, C J; Pittau, M; Leoni, A; Nieddu, A M.
Afiliación
  • Sanna E; Istituto di Patologia Generale, Anatomia Patologica e Clinica Ostetrico-Chirurgica Veterinaria, Facoltà di Veterinaria, Università di Sassari, Italy. patvetss@ssmain.uniss.it
Eur J Histochem ; 44(2): 179-84, 2000.
Article en En | MEDLINE | ID: mdl-10968366
Molecular biological techniques have permitted the rapid and sensitive detection of the Mycobacterium paratuberculosis genome in infected tissues, most commonly by polymerase chain reaction amplification of sequences in the IS900 DNA insertion sequence. The aim of this work was the detection of M. paratuberculosis DNA in ovine tissues by in situ-polymerase chain reaction, which is sensitive and localises the signal within the tissue sample. Paraffin embedded tissues from three acid-fast positive ovine guts with classical lesions of paratuberculosis, and from negative control samples were tested. A 413-bp fragment of the IS900 sequence was amplified in-situ and hybridised to an internal PCR-synthesised digoxygenin-labelled probe. The samples from sheep affected by paratuberculosis clearly showed cell-specific cytoplasmic signals in mucosal and submucosal macrophages. This technique could be useful both in the diagnosis and study of the pathogenesis of infections in which involvement of M. paratuberculosis is suspected.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Paratuberculosis / ADN Bacteriano / Enfermedades de los Bovinos Tipo de estudio: Diagnostic_studies Límite: Animals / Humans Idioma: En Revista: Eur J Histochem Asunto de la revista: HISTOCITOQUIMICA Año: 2000 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Italia
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Paratuberculosis / ADN Bacteriano / Enfermedades de los Bovinos Tipo de estudio: Diagnostic_studies Límite: Animals / Humans Idioma: En Revista: Eur J Histochem Asunto de la revista: HISTOCITOQUIMICA Año: 2000 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Italia