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.
|Title of host publication||PCR Detection of Microbial Pathogens|
|Publication status||Published - 1 Jan 2013|
|Series||Methods in Molecular Biology|
Hansen, W. L. J., Bruggeman, C. A., & Wolffs, P. F. (2013). Pre-analytical sample treatment and DNA extraction protocols for the detection of bacterial pathogens from whole blood. In M. Wilks (Ed.), PCR Detection of Microbial Pathogens (pp. 81-90). Springer. Methods in Molecular Biology, No. 943 https://doi.org/10.1007/978-1-60327-353-4_4