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Fluorescence in situ hybridisation in Carnoy's fixed tonsil tissue.
Clark, S T; Waldvogel-Thurlow, S; Wagner Mackenzie, B; Douglas, R G; Biswas, K.
Afiliação
  • Clark ST; Department of Surgery, Faculty of Medical and Health Sciences, The University of Auckland, Grafton, Auckland, 1023, New Zealand. sitaclark12@gmail.com.
  • Waldvogel-Thurlow S; Department of Surgery, Faculty of Medical and Health Sciences, The University of Auckland, Grafton, Auckland, 1023, New Zealand.
  • Wagner Mackenzie B; Department of Surgery, Faculty of Medical and Health Sciences, The University of Auckland, Grafton, Auckland, 1023, New Zealand.
  • Douglas RG; Department of Surgery, Faculty of Medical and Health Sciences, The University of Auckland, Grafton, Auckland, 1023, New Zealand.
  • Biswas K; Department of Surgery, Faculty of Medical and Health Sciences, The University of Auckland, Grafton, Auckland, 1023, New Zealand.
Sci Rep ; 12(1): 12395, 2022 07 20.
Article em En | MEDLINE | ID: mdl-35858968
ABSTRACT
Fluorescence in situ hybridisation (FISH) is a powerful molecular technique that enables direct visualisation of specific bacterial species. Few studies have established FISH protocols for tonsil tissue in Carnoy's fixative, accordingly limiting its application to investigate the pathogenesis of tonsillar hyperplasia. Tonsil tissue from 24 children undergoing tonsillectomy for either recurrent tonsillitis or sleep-disordered breathing were obtained during a previous study. The specificity of each of the five FISH probes (Fusobacterium spp., Bacteroides spp., Streptococcus spp., Haemophilus influenzae and Pseudomonas spp.) were successfully optimised using pure and mixed bacterial isolates, and in Carnoy's fixed tonsil tissue. Bacteroides spp. were present in 100% of patients with microcolonies. In comparison, the prevalence of Fusobacterium spp. was 93.8%, Streptococcus spp. 85.7%, H. influenzae 82.35% and Pseudomonas spp. 76.5%. Notable differences in the organisation of bacterial taxa within a single microcolony were also observed. This is the first study to establish a robust FISH protocol identifying multiple aerobic and anaerobic bacteria in Carnoy's fixed tonsil tissue. This protocol provides a strong foundation for combining histological and microbiological analyses of Carnoy's fixed tonsil samples. It may also have important implications on the analysis of microorganisms in other human tissues prepared using the same techniques.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Tonsilectomia / Tonsilite Tipo de estudo: Risk_factors_studies Limite: Child / Humans Idioma: En Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Tonsilectomia / Tonsilite Tipo de estudo: Risk_factors_studies Limite: Child / Humans Idioma: En Ano de publicação: 2022 Tipo de documento: Article