Fluorescence in situ hybridisation in Carnoy's fixed tonsil tissue.
Fluorescence in situ hybridisation in Carnoy's fixed tonsil tissue.
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DOI：
10.1038/s41598-022-16309-w
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发表时间：
2022-07-20
影响因子：
4.6
通讯作者：
Biswas, K.
中科院分区：
文献类型：
--
作者：
Clark, S. T.;Waldvogel-Thurlow, S.;Mackenzie, B. Wagner;Douglas, R. G.;Biswas, K.
荧光原位杂交（FISH）是一种强大的分子技术，可以直接观察特定的细菌种类。很少有研究在Carnoy固定物中建立扁桃体组织的FISH方案，因此限制了其在扁桃体增生发病机制研究中的应用。在先前的研究中，从24名因复发性扁桃体炎或睡眠呼吸障碍而接受扁桃体切除术的儿童中获得扁桃体组织。5种FISH探针（梭杆菌、拟杆菌、链球菌、流感嗜血杆菌和假单胞菌）的特异性分别在Carnoy的固定扁桃体组织中使用纯细菌和混合细菌分离物成功优化。100%的微菌落患者中均存在拟杆菌。其中梭杆菌感染率为93.8%，链球菌感染率为85.7%，流感嗜血杆菌感染率为82.35%，假单胞菌感染率为76.5%。在单个微菌落内细菌分类群的组织也观察到显著差异。这是第一个建立一个强大的FISH方案的研究，该方案确定了Carnoy固定扁桃体组织中的多种好氧和厌氧细菌。该方案为结合卡诺伊固定扁桃体样本的组织学和微生物学分析提供了坚实的基础。它也可能对使用相同技术制备的其他人体组织中的微生物分析具有重要意义。
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.
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影响因子：
3.7
作者：
Jensen A;Fagö-Olsen H;Sørensen CH;Kilian M
通讯作者：
Kilian M
影响因子：
9.4
作者：
Hogardt, M;Trebesius, K;Heesemann, J
通讯作者：
Heesemann, J
影响因子：
2.8
作者：
Manz, W;Amann, R;Schleifer, KH
通讯作者：
Schleifer, KH
DOI：
10.1016/j.ijporl.2010.12.014
发表时间：
2011-03-01
影响因子：
1.5
作者：
Kutluhan, Ahmet;Salviz, Mehti;Yesil, Cemile
通讯作者：
Yesil, Cemile
DOI：
10.1007/978-3-662-52959-1_9
发表时间：
2017-01-01
期刊：
FLUORESCENCE IN SITU HYBRIDIZATION (FISH): APPLICATION GUIDE, 2ND EDITION
影响因子：
--
作者：
Liehr, Thomas;Kreskowski, Katharina;Rittscher, Katharina
通讯作者：
Rittscher, Katharina