Role of fungi in chronic rhinosinusitis through ITS sequencing

Objective Next‐generation sequencing increases the sensitivity of fungal identification and may improve our understanding of the role that fungi play in sinus health and disease, which remains incompletely understood. We sequenced the internal transcribed spacer (ITS) amplicon to explore the role of...

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Veröffentlicht in:The Laryngoscope 2018-01, Vol.128 (1), p.16-22
Hauptverfasser: Zhao, Yi Chen, Bassiouni, Ahmed, Tanjararak, Kangsadarn, Vreugde, Sarah, Wormald, Peter‐John, Psaltis, Alkis James
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container_end_page 22
container_issue 1
container_start_page 16
container_title The Laryngoscope
container_volume 128
creator Zhao, Yi Chen
Bassiouni, Ahmed
Tanjararak, Kangsadarn
Vreugde, Sarah
Wormald, Peter‐John
Psaltis, Alkis James
description Objective Next‐generation sequencing increases the sensitivity of fungal identification and may improve our understanding of the role that fungi play in sinus health and disease, which remains incompletely understood. We sequenced the internal transcribed spacer (ITS) amplicon to explore the role of the mycobiome in chronic rhinosinusitis (CRS). Methods Swabs were collected intraoperatively from the middle meatus of 90 patients (63 with CRS; 27 controls). DNA was extracted, and ITS amplicon concentration was measured using fluorometry. Internal transcribed spacer amplicons were sequenced on the Illumina MiSeq (Illumina Inc San Diego CA). Sequencing data were analyzed using QIIME. Results Using conventional detection techniques of culture and histology, fungi only were identified in nine of 63 (14.3%) CRS patients (fungus‐identified group); the remaining 54 CRS patients and all controls did not have fungus identified using the traditional techniques. This fungus‐identified group had a significantly higher average ITS concentration and a significantly lower Shannon's diversity index compared to the other two groups. The most abundant organism sequenced was Aspergillus (35.22% of all sequences). Multivariate analysis showed that positive fungal detection using traditional techniques and computed tomography (CT) double densities were the most important clinical predictors of a high fungal biomass, whereas Lund‐Mackay score, polyps, eosinophilia, and eosinophilic mucus were not significant in comparison. Conclusion Fungal biomass estimated through ITS amplicon concentration correlated with traditional fungal detection techniques and CT double densities. Our results suggest that fungal dysbiosis only occurs in the sinuses of a selected subset of patients, and therefore could not be a universal determinant of sinus disease pathogenesis in all CRS patients. Level of Evidence NA. Laryngoscope, 128:16–22, 2018
doi_str_mv 10.1002/lary.26702
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We sequenced the internal transcribed spacer (ITS) amplicon to explore the role of the mycobiome in chronic rhinosinusitis (CRS). Methods Swabs were collected intraoperatively from the middle meatus of 90 patients (63 with CRS; 27 controls). DNA was extracted, and ITS amplicon concentration was measured using fluorometry. Internal transcribed spacer amplicons were sequenced on the Illumina MiSeq (Illumina Inc San Diego CA). Sequencing data were analyzed using QIIME. Results Using conventional detection techniques of culture and histology, fungi only were identified in nine of 63 (14.3%) CRS patients (fungus‐identified group); the remaining 54 CRS patients and all controls did not have fungus identified using the traditional techniques. This fungus‐identified group had a significantly higher average ITS concentration and a significantly lower Shannon's diversity index compared to the other two groups. The most abundant organism sequenced was Aspergillus (35.22% of all sequences). Multivariate analysis showed that positive fungal detection using traditional techniques and computed tomography (CT) double densities were the most important clinical predictors of a high fungal biomass, whereas Lund‐Mackay score, polyps, eosinophilia, and eosinophilic mucus were not significant in comparison. Conclusion Fungal biomass estimated through ITS amplicon concentration correlated with traditional fungal detection techniques and CT double densities. Our results suggest that fungal dysbiosis only occurs in the sinuses of a selected subset of patients, and therefore could not be a universal determinant of sinus disease pathogenesis in all CRS patients. Level of Evidence NA. 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We sequenced the internal transcribed spacer (ITS) amplicon to explore the role of the mycobiome in chronic rhinosinusitis (CRS). Methods Swabs were collected intraoperatively from the middle meatus of 90 patients (63 with CRS; 27 controls). DNA was extracted, and ITS amplicon concentration was measured using fluorometry. Internal transcribed spacer amplicons were sequenced on the Illumina MiSeq (Illumina Inc San Diego CA). Sequencing data were analyzed using QIIME. Results Using conventional detection techniques of culture and histology, fungi only were identified in nine of 63 (14.3%) CRS patients (fungus‐identified group); the remaining 54 CRS patients and all controls did not have fungus identified using the traditional techniques. This fungus‐identified group had a significantly higher average ITS concentration and a significantly lower Shannon's diversity index compared to the other two groups. The most abundant organism sequenced was Aspergillus (35.22% of all sequences). Multivariate analysis showed that positive fungal detection using traditional techniques and computed tomography (CT) double densities were the most important clinical predictors of a high fungal biomass, whereas Lund‐Mackay score, polyps, eosinophilia, and eosinophilic mucus were not significant in comparison. Conclusion Fungal biomass estimated through ITS amplicon concentration correlated with traditional fungal detection techniques and CT double densities. Our results suggest that fungal dysbiosis only occurs in the sinuses of a selected subset of patients, and therefore could not be a universal determinant of sinus disease pathogenesis in all CRS patients. Level of Evidence NA. 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subjects Biomass
Chronic Disease
chronic rhinosinusitis
DNA, Fungal - analysis
Endoscopy
Female
fungal microbiome
Fungi
Humans
Male
Microbiota
Multivariate analysis
Mycoses - microbiology
Nasal Mucosa - microbiology
Polymerase Chain Reaction
Prospective Studies
Rhinitis
Rhinitis - microbiology
Sinonasal microbiome
sinonasal mycobiome
Sinusitis
Sinusitis - microbiology
title Role of fungi in chronic rhinosinusitis through ITS sequencing
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