Fluorescence in situ hybridisation in Carnoy's fixed tonsil tissue.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
20 07 2022
20 07 2022
Historique:
received:
24
02
2022
accepted:
07
07
2022
entrez:
20
7
2022
pubmed:
21
7
2022
medline:
23
7
2022
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 35858968
doi: 10.1038/s41598-022-16309-w
pii: 10.1038/s41598-022-16309-w
pmc: PMC9300673
doi:
Substances chimiques
Fixatives
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
12395Informations de copyright
© 2022. The Author(s).
Références
Postgrad Med J. 2018 Jul;94(1113):398-403
pubmed: 29884749
Respir Med. 1993 May;87(4):303-8
pubmed: 9728232
J Clin Microbiol. 2000 Feb;38(2):818-25
pubmed: 10655391
J Clin Microbiol. 2011 Apr;49(4):1411-20
pubmed: 21307211
Int J Pediatr Otorhinolaryngol. 2005 Jan;69(1):9-19
pubmed: 15627441
J Vis Exp. 2019 Oct 18;(152):
pubmed: 31680675
Ear Nose Throat J. 2017 Jun;96(6):E29-E32
pubmed: 28636739
Sci Rep. 2021 Nov 9;11(1):21940
pubmed: 34753993
J Clin Pathol. 2007 Mar;60(3):253-60
pubmed: 16698947
Respir Med. 1990 Sep;84(5):395-400
pubmed: 2247668
Int J Pediatr Otorhinolaryngol. 2018 Oct;113:281-288
pubmed: 30174001
Cell Microbiol. 2009 Jul;11(7):1034-43
pubmed: 19374653
Environ Microbiol. 2006 May;8(5):871-84
pubmed: 16623744
J Clin Microbiol. 2000 Feb;38(2):830-8
pubmed: 10655393
Appl Environ Microbiol. 1998 Sep;64(9):3336-45
pubmed: 9726880
J Breath Res. 2017 Sep 06;11(3):036010
pubmed: 28875948
Curr Microbiol. 2014 Jan;68(1):21-9
pubmed: 23934353
Arch Otolaryngol. 1984 Dec;110(12):803-5
pubmed: 6334513
Microbiology (Reading). 1996 May;142 ( Pt 5):1097-1106
pubmed: 8704951
Appl Environ Microbiol. 1990 Jun;56(6):1919-25
pubmed: 2200342
Methods Mol Biol. 2009;493:191-213
pubmed: 18839349
PLoS One. 2010 Mar 01;5(3):e9452
pubmed: 20209109
Int J Pediatr Otorhinolaryngol. 2011 Mar;75(3):391-4
pubmed: 21227516
Microbiology (Reading). 2003 May;149(Pt 5):1095-1102
pubmed: 12724371
Environ Microbiol. 2013 Feb;15(2):535-47
pubmed: 23113966
Nephrol Dial Transplant. 2017 Dec 01;32(12):2072-2079
pubmed: 27683270
Br J Biomed Sci. 2005;62(2):66-70
pubmed: 15997879
Eur Arch Otorhinolaryngol. 2017 Aug;274(8):3153-3159
pubmed: 28551703
Clin Infect Dis. 1995 Jul;21(1):171-6
pubmed: 7578726
Eur J Clin Microbiol Infect Dis. 2016 Jun;35(6):963-70
pubmed: 27025724
Laryngoscope. 2020 Dec;130(12):E791-E794
pubmed: 31747474
Cureus. 2018 Sep 22;10(9):e3343
pubmed: 30473976
Int J Pediatr Otorhinolaryngol. 2022 Jun;157:111128
pubmed: 35421675
J Infect Dis. 2007 Oct 1;196(7):1080-7
pubmed: 17763332
JAMA. 1980 Oct 10;244(15):1696-8
pubmed: 7411827
Int J Pediatr Otorhinolaryngol. 2009 Sep;73(9):1242-8
pubmed: 19525016
Science. 1999 May 21;284(5418):1318-22
pubmed: 10334980
Annu Rev Microbiol. 2003;57:677-701
pubmed: 14527295
PLoS One. 2013;8(2):e56418
pubmed: 23437130