Ascitic microbiota composition is correlated with clinical severity in cirrhosis with portal hypertension
Ascitic microbiota composition is correlated with clinical severity in cirrhosis with portal hypertension
Identification of pathogenic bacteria in ascites correlates with poor clinical outcomes. Ascites samples are commonly reported culture-negative, even where frank infection is indicated. Culture-independent methods have previously reported bacterial DNA in ascites, however, whether this represents viable bacterial populations has not been determined. We report the first application of 16S rRNA gene pyrosequencing and quantitative PCR in conjunction with propidium monoazide sample treatment to characterise the viable bacterial composition of ascites. Twenty five cirrhotic patients undergoing paracentesis provided ascites. Samples were treated with propidium monoazide to exclude non-viable bacterial DNA. Total bacterial load was quantified by 16S rRNA Q-PCR with species identity and relative abundance determined by 16S rRNA gene pyrosequencing. Correlation of molecular microbiology data with clinical measures and diagnostic microbiology was performed. Viable bacterial signal was obtained in 84% of ascites samples, both by Q-PCR and pyrosequencing. Approximately 190,000 ribosomal pyrosequences were obtained, representing 236 species, including both gut and non gut-associated species. Substantial variation in the species detected was observed between patients. Statistically significant relationships were identified between the bacterial community similarity and clinical measures, including ascitic polymorphonuclear leukocyte count and Child-Pugh class. Viable bacteria are present in the ascites of a majority of patients with cirrhosis including those with no clinical signs of infection. Microbiota composition significantly correlates with clinical measures. Entry of bacteria into ascites is unlikely to be limited to translocation from the gut, raising fundamental questions about the processes that underlie the development of spontaneous bacterial peritonitis.
e74884
Rogers, G.B.
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van der Gast, C.J.
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Bruce, K.D.
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Marsh, P.
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Collins, J.E.
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Sutton, J.
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Wright, M.
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25 September 2013
Rogers, G.B.
9a939cbd-f83a-41ce-8ba4-a71065e0b87e
van der Gast, C.J.
332ec7c1-c2c9-4d5e-a935-9e170980dfdb
Bruce, K.D.
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Marsh, P.
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Collins, J.E.
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Sutton, J.
0968e5f3-7c74-4e97-a330-c6fe21c0a25e
Wright, M.
43325ef9-3459-4c75-b3bf-cf8d8dac2a21
Rogers, G.B., van der Gast, C.J., Bruce, K.D., Marsh, P., Collins, J.E., Sutton, J. and Wright, M.
(2013)
Ascitic microbiota composition is correlated with clinical severity in cirrhosis with portal hypertension.
PLoS ONE, 8 (9), .
(doi:10.1371/journal.pone.0074884).
(PMID:24086392)
Abstract
Identification of pathogenic bacteria in ascites correlates with poor clinical outcomes. Ascites samples are commonly reported culture-negative, even where frank infection is indicated. Culture-independent methods have previously reported bacterial DNA in ascites, however, whether this represents viable bacterial populations has not been determined. We report the first application of 16S rRNA gene pyrosequencing and quantitative PCR in conjunction with propidium monoazide sample treatment to characterise the viable bacterial composition of ascites. Twenty five cirrhotic patients undergoing paracentesis provided ascites. Samples were treated with propidium monoazide to exclude non-viable bacterial DNA. Total bacterial load was quantified by 16S rRNA Q-PCR with species identity and relative abundance determined by 16S rRNA gene pyrosequencing. Correlation of molecular microbiology data with clinical measures and diagnostic microbiology was performed. Viable bacterial signal was obtained in 84% of ascites samples, both by Q-PCR and pyrosequencing. Approximately 190,000 ribosomal pyrosequences were obtained, representing 236 species, including both gut and non gut-associated species. Substantial variation in the species detected was observed between patients. Statistically significant relationships were identified between the bacterial community similarity and clinical measures, including ascitic polymorphonuclear leukocyte count and Child-Pugh class. Viable bacteria are present in the ascites of a majority of patients with cirrhosis including those with no clinical signs of infection. Microbiota composition significantly correlates with clinical measures. Entry of bacteria into ascites is unlikely to be limited to translocation from the gut, raising fundamental questions about the processes that underlie the development of spontaneous bacterial peritonitis.
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Published date: 25 September 2013
Organisations:
Clinical & Experimental Sciences
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Local EPrints ID: 358215
URI: http://eprints.soton.ac.uk/id/eprint/358215
ISSN: 1932-6203
PURE UUID: d9911297-793c-45d9-8ce3-9f0443911b0c
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Date deposited: 09 Oct 2013 12:33
Last modified: 14 Mar 2024 15:02
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Author:
G.B. Rogers
Author:
C.J. van der Gast
Author:
K.D. Bruce
Author:
P. Marsh
Author:
J. Sutton
Author:
M. Wright
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