Reference : Detection of bacteria in sterile body fluids

	Document type :	Scientific congresses and symposiums : Poster
	Discipline(s) :	Human health sciences : Immunology & infectious disease Human health sciences : Laboratory medicine & medical technology
	To cite this reference:	http://hdl.handle.net/2268/84112

Title :	Detection of bacteria in sterile body fluids
Language :	English
Alternative title :	[en] Détection de bactéries dans les fluides corporels stériles
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 >]
Publication date :	Oct-1998
Page number :	1x1.5
On invitation :	No
Audience :	National
Event name :	Symposium national
Event date :	17 OCTOBRE 1998
Event organizer :	Société belge de biologie clinique & Société belge de chimie clinique
Event place (city) :	Leuven
Event country :	BELGIUM
Keywords :	[en] 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).
Target :	Researchers ; Professionals ; Students
Permalink :	http://hdl.handle.net/2268/84112

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