Characterisation of bacteria in ascites - reporting the potential of culture-independent, molecular analysis

Geraint Rogers, L. Russel, P. Preston, Peter Marsh, Jane Collins, J. Saunders, Julian Sutton, David Fine, Kenneth Bruce, Mark Wright

    Research output: Contribution to journalArticlepeer-review

    22 Citations (Scopus)

    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 × 102 to 5.4 × 107 cfu/ml, with a mean value of 1.9 × 106 cfu/ml (standard deviation ± 9.6 × 106 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.

    Original languageEnglish
    Pages (from-to)533-541
    Number of pages9
    JournalEuropean Journal of Clinical Microbiology and Infectious Diseases
    Volume29
    Issue number5
    DOIs
    Publication statusPublished - May 2010

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