title: Pre-analytical sample treatment and DNA extraction protocols for the detection of bacterial pathogens from whole blood.
abstract: Molecular diagnostics is an increasing popular approach for the direct detection and identification of pathogenic bacteria in clinical samples. Conventional culture techniques are time-consuming and therefore causing a delay in the diagnosis of the patient. Alternative techniques based on nucleic acid amplification offer a shorter turn-around-time and the ability to identify fastidious and non-cultivable organisms. However, molecular detection of bacteria in blood, by for example PCR, RT-PCR, or sequencing of the 16S rDNA genes is often complicated by the presence of PCR-inhibitory compounds. Here we describe several different methods for the extraction of bacterial DNA from whole blood samples. The methods differ regarding costs, hands-on time as well as regarding sensitivity. In combination with a model PCR the detection limits that can be reached using the different methods range from 1,000 to 50 cfu/ml.
authors.affiliation: Department of Medical Microbiology, Maastricht University Medical Center, Maastricht, The Netherlands.
authors.forename: Wendy L J
authors.initials: WL
authors.lastname: Hansen
authors.forename: Cathrien A
authors.initials: CA
authors.lastname: Bruggeman
authors.forename: Petra F G
authors.initials: PF
authors.lastname: Wolffs
mesh_terms: D001419:Bacteria / Q000235:genetics*; D001424:Bacterial Infections / Q000175:diagnosis*; D017848:Blood-Borne Pathogens; D004269:DNA, Bacterial / Q000302:isolation & purification*; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial