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Poster (Scientific congresses and symposiums)
Detection of bacteria in sterile body fluids
CHRISTIAENS, Geneviève; HAYETTE, Marie-Pierre; De Mol, Patrick
1998Symposium national


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Keywords :
bacteria; sterile body fluids; PCR
Abstract :
[en] Purpose of the study: Development of a polymerase chain reaction (PCR) method for the detection of any bacterial DNA in synovial, cerebrospinal, pleural, and peritoneal fluids. Methods: Most of bacterial species possess highly conserved, multicopy 16S ribosomal RNA genes, which can be hybridized with a single set of oligonucleotide primers. Samples were processed by an extraction protocol using proteinase K, and subjected to PCR amplification using two universal bacterial primers: RW01 and DG74; then the PCR products were detected by ethidium bromide gel electrophoresis. Study: Synovial, cerebrospinal, pleural and peritoneal fluids, obtained from 32 patients were analyzed by Gram stain, culture and PCR. Results: 1. The limit of detection, determined by analyzing successive dilutions of Staphylococcus aureus and Escherichia coli cultures in sterile water, was: 1.105 cfu/100µl. 2. We obtained 9 positive samples by culture: - 7 synovial fluids: S. agalactiae, S. viridans, coagulase negative Staphylococcus (2), S. epidermidis (2), and S. aureus. - 2 pleural fluids: S. pyogenes and Enterobacter aerogenes. All were PCR positive. 3. We tested 23 culture negative samples. All were negative by PCR. Conclusion: PCR presents some interesting features: 1. Only small amount of sample is necessary (100µl). 2. The duration of the test is shorter than 8 hours. 3. PCR provides similar or eventually greater sensitivity than culture and an excellent specificity (no false-positive and no false-negative results actually observed).
Disciplines :
Immunology & infectious disease
Laboratory medicine & medical technology
Author, co-author :
CHRISTIAENS, Geneviève ;  Centre Hospitalier Universitaire de Liège - CHU > Direction médicale
HAYETTE, Marie-Pierre ;  Centre Hospitalier Universitaire de Liège - CHU > Microbiologie médicale
De Mol, Patrick ;  Centre Hospitalier Universitaire de Liège - CHU > Microbiologie médicale
Language :
Title :
Detection of bacteria in sterile body fluids
Alternative titles :
[en] Détection de bactéries dans les fluides corporels stériles
Publication date :
October 1998
Number of pages :
Event name :
Symposium national
Event organizer :
Société belge de biologie clinique & Société belge de chimie clinique
Event place :
Leuven, Belgium
Event date :
17 OCTOBRE 1998
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since 21 February 2011


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