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In situ-PCR for the detection of Mycobacterium paratuberculosis DNA in paraffin-embedded tissues

Sanna, Ennio and Woodall, Christopher and Watt, N. J. and Clarke, C. J. and Pittau, M. and Leoni, Antonio and Nieddu, Antonio Mario S. (2000) In situ-PCR for the detection of Mycobacterium paratuberculosis DNA in paraffin-embedded tissues. European Journal of Histochemistry, Vol. 44 (2), p. 179-184. ISSN 1121-760X. Article.

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Abstract

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.

Item Type:Article
ID Code:1169
Status:Published
Refereed:Yes
Uncontrolled Keywords:Mycobacterium paratuberculosis, PCR
Subjects:Area 07 - Scienze agrarie e veterinarie > VET/03 Patologia generale e anatomia patologica veterinaria
Divisions:001 Università di Sassari > 01 Dipartimenti > Patologia e clinica veterinaria
Publisher:Luigi Ponzio
ISSN:1121-760X
Deposited On:18 Aug 2009 10:04

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