|Reference : Detection of bacteria in sterile body fluids|
|Scientific congresses and symposiums : Poster|
|Human health sciences : Immunology & infectious disease|
Human health sciences : Laboratory medicine & medical technology
|Detection of bacteria in sterile body fluids|
|[en] Détection de bactéries dans les fluides corporels stériles|
|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 >]|
|17 OCTOBRE 1998|
|Société belge de biologie clinique & Société belge de chimie clinique|
|[en] bacteria ; sterile body fluids ; PCR|
|[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
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.
Synovial, cerebrospinal, pleural and peritoneal fluids, obtained from
32 patients were analyzed by Gram stain, culture and PCR.
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.
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).
|Researchers ; Professionals ; Students|
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