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 |
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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 |
format | Article |
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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</description><identifier>ISSN: 0023-852X</identifier><identifier>EISSN: 1531-4995</identifier><identifier>DOI: 10.1002/lary.26702</identifier><identifier>PMID: 28675446</identifier><language>eng</language><publisher>United States: Wiley Subscription Services, Inc</publisher><subject>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</subject><ispartof>The Laryngoscope, 2018-01, Vol.128 (1), p.16-22</ispartof><rights>2017 The American Laryngological, Rhinological and Otological Society, Inc.</rights><rights>2018 The American Laryngological, Rhinological and Otological Society, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4232-70ffcadb8cff5254c78f21a54e41ef131ee09650887919b6c989d27aaf631fc03</citedby><cites>FETCH-LOGICAL-c4232-70ffcadb8cff5254c78f21a54e41ef131ee09650887919b6c989d27aaf631fc03</cites><orcidid>0000-0002-5545-0194</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Flary.26702$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Flary.26702$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28675446$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhao, Yi Chen</creatorcontrib><creatorcontrib>Bassiouni, Ahmed</creatorcontrib><creatorcontrib>Tanjararak, Kangsadarn</creatorcontrib><creatorcontrib>Vreugde, Sarah</creatorcontrib><creatorcontrib>Wormald, Peter‐John</creatorcontrib><creatorcontrib>Psaltis, Alkis James</creatorcontrib><title>Role of fungi in chronic rhinosinusitis through ITS sequencing</title><title>The Laryngoscope</title><addtitle>Laryngoscope</addtitle><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</description><subject>Biomass</subject><subject>Chronic Disease</subject><subject>chronic rhinosinusitis</subject><subject>DNA, Fungal - analysis</subject><subject>Endoscopy</subject><subject>Female</subject><subject>fungal microbiome</subject><subject>Fungi</subject><subject>Humans</subject><subject>Male</subject><subject>Microbiota</subject><subject>Multivariate analysis</subject><subject>Mycoses - microbiology</subject><subject>Nasal Mucosa - microbiology</subject><subject>Polymerase Chain Reaction</subject><subject>Prospective Studies</subject><subject>Rhinitis</subject><subject>Rhinitis - microbiology</subject><subject>Sinonasal microbiome</subject><subject>sinonasal mycobiome</subject><subject>Sinusitis</subject><subject>Sinusitis - microbiology</subject><issn>0023-852X</issn><issn>1531-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1LAzEQhoMotlYv_gBZ8CLC1kw22SQXoRS_oCDUCnpatmnSpmyzNeki_fembvXgwdPAzMM7Mw9C54D7gDG5qUq_7ZOcY3KAusAySKmU7BB14zBLBSNvHXQSwhJj4BnDx6hDRM4ZpXkX3Y7rSie1SUzj5jaxLlELXzurEr-wrg7WNcFubEg2sd3MF8nT5CUJ-qPRTlk3P0VHpqyCPtvXHnq9v5sMH9PR88PTcDBKFSUZSTk2RpWzqVDGMMKo4sIQKBnVFLSBDLTGMmdYCC5BTnMlhZwRXpYmz8AonPXQVZu79nXcHTbFygalq6p0um5CARKYiJ_CDr38gy7rxrt4XaS4yGkOkkTquqWUr0Pw2hRrb1dRZAG42FktdlaLb6sRvthHNtOVnv2iPxojAC3waSu9_SeqGA3G723oF4tHgSE</recordid><startdate>201801</startdate><enddate>201801</enddate><creator>Zhao, Yi Chen</creator><creator>Bassiouni, Ahmed</creator><creator>Tanjararak, Kangsadarn</creator><creator>Vreugde, Sarah</creator><creator>Wormald, Peter‐John</creator><creator>Psaltis, Alkis James</creator><general>Wiley Subscription Services, Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>K9.</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-5545-0194</orcidid></search><sort><creationdate>201801</creationdate><title>Role of fungi in chronic rhinosinusitis through ITS sequencing</title><author>Zhao, Yi Chen ; Bassiouni, Ahmed ; Tanjararak, Kangsadarn ; Vreugde, Sarah ; Wormald, Peter‐John ; Psaltis, Alkis James</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4232-70ffcadb8cff5254c78f21a54e41ef131ee09650887919b6c989d27aaf631fc03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Biomass</topic><topic>Chronic Disease</topic><topic>chronic rhinosinusitis</topic><topic>DNA, Fungal - analysis</topic><topic>Endoscopy</topic><topic>Female</topic><topic>fungal microbiome</topic><topic>Fungi</topic><topic>Humans</topic><topic>Male</topic><topic>Microbiota</topic><topic>Multivariate analysis</topic><topic>Mycoses - microbiology</topic><topic>Nasal Mucosa - microbiology</topic><topic>Polymerase Chain Reaction</topic><topic>Prospective Studies</topic><topic>Rhinitis</topic><topic>Rhinitis - microbiology</topic><topic>Sinonasal microbiome</topic><topic>sinonasal mycobiome</topic><topic>Sinusitis</topic><topic>Sinusitis - microbiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhao, Yi Chen</creatorcontrib><creatorcontrib>Bassiouni, Ahmed</creatorcontrib><creatorcontrib>Tanjararak, Kangsadarn</creatorcontrib><creatorcontrib>Vreugde, Sarah</creatorcontrib><creatorcontrib>Wormald, Peter‐John</creatorcontrib><creatorcontrib>Psaltis, Alkis James</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><jtitle>The Laryngoscope</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhao, Yi Chen</au><au>Bassiouni, Ahmed</au><au>Tanjararak, Kangsadarn</au><au>Vreugde, Sarah</au><au>Wormald, Peter‐John</au><au>Psaltis, Alkis James</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Role of fungi in chronic rhinosinusitis through ITS sequencing</atitle><jtitle>The Laryngoscope</jtitle><addtitle>Laryngoscope</addtitle><date>2018-01</date><risdate>2018</risdate><volume>128</volume><issue>1</issue><spage>16</spage><epage>22</epage><pages>16-22</pages><issn>0023-852X</issn><eissn>1531-4995</eissn><abstract>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</abstract><cop>United States</cop><pub>Wiley Subscription Services, Inc</pub><pmid>28675446</pmid><doi>10.1002/lary.26702</doi><tpages>7</tpages><orcidid>https://orcid.org/0000-0002-5545-0194</orcidid></addata></record> |
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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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