Visualisation of Mycobacterium avium subsp. paratuberculosis in cultured cells, infected sheep and human tissue sections using fluorescent in situ hybridization (FISH)

• Published in: Journal of microbiological methods 2024-09, Vol.224, p.107001, Article 107001
• Main Authors: Rayment, Neil, Rhodes, Glenn, Hudspith, Barry, Hughes, Valerie, Chianini, Francesca, Agrawal, Gaurav, Bull, Tim J., Pickup, Roger, Sanderson, Jeremy
• Format: Article
• Language: English
• Subjects: Animals; Cells, Cultured; Crohn's disease; FISH; Humans; In Situ Hybridization, Fluorescence - methods; Johne's disease; Microscopy, Confocal - methods; Mycobacterium avium ssp. paratuberculosis (MAP); Mycobacterium avium subsp. paratuberculosis - genetics; Mycobacterium avium subsp. paratuberculosis - isolation & purification; Oligonucleotide Probes - genetics; Paratuberculosis - diagnosis; Paratuberculosis - microbiology; Sensitivity and Specificity; Sheep; Sheep Diseases - diagnosis; Sheep Diseases - microbiology
• ISSN: 0167-7012; 1872-8359; 1872-8359
• DOI: 10.1016/j.mimet.2024.107001

We describe the development, testing and specificity of a modified oligonucleotide probe for the specific detection of Mycobacterium avium subsp. paratuberculosis (MAP) in culture and in infected tissue using fluorescent in situ hybridisation and confocal microscopy. The detection of MAP in both animal and human tissue using our modified probe allows for a more rapid diagnosis of MAP infection compared to the more often applied detection methods of culture and PCR and has the potential for quantification of cellular abundance. This approach would enable earlier treatment intervention and therefore the potential for reduced morbidity.