Characterization of bacterial community diversity in chronic rhinosinusitis infections using novel culture-independent techniques
Characterization of bacterial community diversity in chronic rhinosinusitis infections using novel culture-independent techniques
BACKGROUND: Chronic rhinosinusitis (CRS) with or without polyps is a common chronic upper airway condition of multifactorial origin. Fundamental to effective treatment of any infection is the ability to accurately characterize the underlying cause. Many studies have shown that only a small fraction of the total range of bacterial species present in CRS is detected through conventional culture-dependent techniques. Consequently, culture data are often unrepresentative of the true diversity of the microbial community within the sample. These drawbacks, along with the length of time required to complete the analysis, strongly support the development of alternative means of assessing which bacterial species are present. As such, molecular microbiological approaches that assess the content of clinical samples in a culture-independent manner could significantly enhance the range and quality of data obtained routinely from such samples. We aimed to characterize the bacterial diversity present in tissue and mucus samples taken from the CRS setting using molecular nonculture-dependent techniques.
METHODS: Through 16S ribosomal RNA (rRNA) gene clone sequencing and terminal restriction fragment length polymorphism (T-RFLP) analysis, the bacteria present in 70 clinical samples from 43 CRS patients undergoing endoscopic sinus surgery were characterized.
RESULTS: Bacterial T-RFLP profiles were generated for 70 of 73 samples and a total of 48 separate bands were detected. Species belonging to 34 genera were identified as present by clone sequence analysis. Of the species detected, those within the genera Pseudomonas, Citrobacter, Haemophilus, Propionibacterium, Staphylococcus, and Streptococcus were found numerically dominant, with Pseudomonas aeruginosa the most frequently detected species.
CONCLUSION: This study has validated the use of the culture-independent technique T-RFLP in sinonasal samples. Preliminary characterization of the microbial diversity in CRS suggests a complex range of common and novel bacterial species within the upper airway in CRS, providing further evidence for the polymicrobial etiology of CRS.
Adult, Chronic Disease, Feasibility Studies, Female, Humans, Male, Middle Aged, Molecular Typing/methods, Paranasal Sinuses/microbiology, Polymorphism, Restriction Fragment Length, Pseudomonas Infections/complications, Pseudomonas aeruginosa/genetics, RNA, Ribosomal, 16S/analysis, Rhinitis/etiology, Sinusitis/etiology
e133-e140
Stressmann, Franziska A
4d9bc3a9-dfc5-4957-9888-d4e245ab139a
Rogers, Geraint B
bd90d82a-4150-4e01-a755-b81559586da2
Chan, Samuel W
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Howarth, Peter H
ff19c8c4-86b0-4a88-8f76-b3d87f142a21
Harries, Philip G
df33be64-b580-4fdf-ac4d-4c842b55d753
Bruce, Ken D
e4f5f6c6-3041-4c04-9e73-11d2d76c1d2a
Salib, Rami J
d6fde1c1-5b5e-43f7-ae1c-42cce6a0c9fc
2011
Stressmann, Franziska A
4d9bc3a9-dfc5-4957-9888-d4e245ab139a
Rogers, Geraint B
bd90d82a-4150-4e01-a755-b81559586da2
Chan, Samuel W
3ddf4402-7636-4b05-9f63-c7aa01f7390c
Howarth, Peter H
ff19c8c4-86b0-4a88-8f76-b3d87f142a21
Harries, Philip G
df33be64-b580-4fdf-ac4d-4c842b55d753
Bruce, Ken D
e4f5f6c6-3041-4c04-9e73-11d2d76c1d2a
Salib, Rami J
d6fde1c1-5b5e-43f7-ae1c-42cce6a0c9fc
Stressmann, Franziska A, Rogers, Geraint B, Chan, Samuel W, Howarth, Peter H, Harries, Philip G, Bruce, Ken D and Salib, Rami J
(2011)
Characterization of bacterial community diversity in chronic rhinosinusitis infections using novel culture-independent techniques.
American journal of rhinology & allergy, 25 (4), .
(doi:10.2500/ajra.2011.25.3628).
Abstract
BACKGROUND: Chronic rhinosinusitis (CRS) with or without polyps is a common chronic upper airway condition of multifactorial origin. Fundamental to effective treatment of any infection is the ability to accurately characterize the underlying cause. Many studies have shown that only a small fraction of the total range of bacterial species present in CRS is detected through conventional culture-dependent techniques. Consequently, culture data are often unrepresentative of the true diversity of the microbial community within the sample. These drawbacks, along with the length of time required to complete the analysis, strongly support the development of alternative means of assessing which bacterial species are present. As such, molecular microbiological approaches that assess the content of clinical samples in a culture-independent manner could significantly enhance the range and quality of data obtained routinely from such samples. We aimed to characterize the bacterial diversity present in tissue and mucus samples taken from the CRS setting using molecular nonculture-dependent techniques.
METHODS: Through 16S ribosomal RNA (rRNA) gene clone sequencing and terminal restriction fragment length polymorphism (T-RFLP) analysis, the bacteria present in 70 clinical samples from 43 CRS patients undergoing endoscopic sinus surgery were characterized.
RESULTS: Bacterial T-RFLP profiles were generated for 70 of 73 samples and a total of 48 separate bands were detected. Species belonging to 34 genera were identified as present by clone sequence analysis. Of the species detected, those within the genera Pseudomonas, Citrobacter, Haemophilus, Propionibacterium, Staphylococcus, and Streptococcus were found numerically dominant, with Pseudomonas aeruginosa the most frequently detected species.
CONCLUSION: This study has validated the use of the culture-independent technique T-RFLP in sinonasal samples. Preliminary characterization of the microbial diversity in CRS suggests a complex range of common and novel bacterial species within the upper airway in CRS, providing further evidence for the polymicrobial etiology of CRS.
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More information
Published date: 2011
Keywords:
Adult, Chronic Disease, Feasibility Studies, Female, Humans, Male, Middle Aged, Molecular Typing/methods, Paranasal Sinuses/microbiology, Polymorphism, Restriction Fragment Length, Pseudomonas Infections/complications, Pseudomonas aeruginosa/genetics, RNA, Ribosomal, 16S/analysis, Rhinitis/etiology, Sinusitis/etiology
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Local EPrints ID: 503745
URI: http://eprints.soton.ac.uk/id/eprint/503745
ISSN: 1945-8932
PURE UUID: 75d8a571-4cd5-4cad-9ef1-7627907cc2e9
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Date deposited: 12 Aug 2025 16:52
Last modified: 13 Aug 2025 02:00
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Author:
Franziska A Stressmann
Author:
Geraint B Rogers
Author:
Samuel W Chan
Author:
Philip G Harries
Author:
Ken D Bruce
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