Characterisation of bacteria in ascites - reporting the potential of culture-independent, molecular analysis.
Characterisation of bacteria in ascites - reporting the potential of culture-independent, molecular analysis.
Spontaneous bacterial peritonitis (SBP) is a severe complication of liver disease. A significant proportion of patients have culture-negative ascites, despite having similar signs, symptoms and mortality to those with SBP. Therefore, empirical antibiotic treatment for infection is often started without knowledge of the causative organisms.
Here, we investigated the potential of molecular techniques to provide rapid and accurate characterisation of the bacteria present in ascitic fluid. Ascites samples were obtained from 29 cirrhotic patients undergoing clinically indicated therapeutic paracentesis. Bacterial content was determined by terminal restriction fragment length polymorphism (T-RFLP) analysis, quantitative polymerase chain reaction (PCR) and 16S ribosomal clone sequence analysis. Bacterial signal was detected in all samples, compared to three out of ten using standard methods.
Bacterial loads ranged from 5.5 x 10(2) to 5.4 x 10(7) cfu/ml, with a mean value of 1.9 x 10(6) cfu/ml (standard deviation +/- 9.6 x 10(6) cfu/ml). In all but one instance, bacterial species identified by culture were also confirmed by molecular analyses. Preliminary data presented here suggests that culture-independent, molecular analyses could provide rapid characterisation of the bacterial content of ascites fluid, providing a basis for the investigation of SBP development and allowing early and targeted antibiotic intervention.
533-541
Rogers, G.B.
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Russell, L.E.
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Preston, P.G.
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Marsh, P.
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Collins, J.E.
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Saunders, J.
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Sutton, J.
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Fine, D.
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Bruce, K.D.
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Wright, M.
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May 2010
Rogers, G.B.
9a939cbd-f83a-41ce-8ba4-a71065e0b87e
Russell, L.E.
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Preston, P.G.
386b5b54-db3c-4062-9a9d-014c4fdfd5f9
Marsh, P.
28ca4a02-c63e-45bb-8288-52806daad572
Collins, J.E.
be0e66f1-3036-47fa-9d7e-914c48710ba4
Saunders, J.
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Sutton, J.
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Fine, D.
ce3d6a77-040e-4aec-a8f5-4c4c22431605
Bruce, K.D.
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Wright, M.
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Rogers, G.B., Russell, L.E., Preston, P.G., Marsh, P., Collins, J.E., Saunders, J., Sutton, J., Fine, D., Bruce, K.D. and Wright, M.
(2010)
Characterisation of bacteria in ascites - reporting the potential of culture-independent, molecular analysis.
European Journal of Clinical Microbiology & Infectious Diseases, 29 (5), .
(doi:10.1007/s10096-010-0891-5).
(PMID:20238135)
Abstract
Spontaneous bacterial peritonitis (SBP) is a severe complication of liver disease. A significant proportion of patients have culture-negative ascites, despite having similar signs, symptoms and mortality to those with SBP. Therefore, empirical antibiotic treatment for infection is often started without knowledge of the causative organisms.
Here, we investigated the potential of molecular techniques to provide rapid and accurate characterisation of the bacteria present in ascitic fluid. Ascites samples were obtained from 29 cirrhotic patients undergoing clinically indicated therapeutic paracentesis. Bacterial content was determined by terminal restriction fragment length polymorphism (T-RFLP) analysis, quantitative polymerase chain reaction (PCR) and 16S ribosomal clone sequence analysis. Bacterial signal was detected in all samples, compared to three out of ten using standard methods.
Bacterial loads ranged from 5.5 x 10(2) to 5.4 x 10(7) cfu/ml, with a mean value of 1.9 x 10(6) cfu/ml (standard deviation +/- 9.6 x 10(6) cfu/ml). In all but one instance, bacterial species identified by culture were also confirmed by molecular analyses. Preliminary data presented here suggests that culture-independent, molecular analyses could provide rapid characterisation of the bacterial content of ascites fluid, providing a basis for the investigation of SBP development and allowing early and targeted antibiotic intervention.
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Published date: May 2010
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Local EPrints ID: 155221
URI: http://eprints.soton.ac.uk/id/eprint/155221
ISSN: 0934-9723
PURE UUID: 63541218-af1e-4580-85f3-74c391dd3dd8
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Date deposited: 28 May 2010 10:06
Last modified: 14 Mar 2024 01:37
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Author:
G.B. Rogers
Author:
L.E. Russell
Author:
P.G. Preston
Author:
P. Marsh
Author:
J. Saunders
Author:
J. Sutton
Author:
D. Fine
Author:
K.D. Bruce
Author:
M. Wright
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