The DNA was isolated from blood specimens using the QIAamp DNA Blood Midi Kit (Qiagen).
DNA was prepared from tissue and blood specimens using the conventional proteinase K digestion and phenol/chloroform extraction method.
DNA was extracted from whole blood specimens using the MagNA Pure LC 2.0 instrument and DNA Isolation kit III for bacteria (Roche, Mannheim, Germany) and were tested for the S. pneumoniae lytA gene by a quantitative real-time PCR [ 17].
We rapidly identified extended-spectrum β-lactamase (ESBL) producers prospectively among 245 gram-negative bacilli positive cultured blood specimens using the Rapid ESBL Nordmann/Dortet/Poirel test and direct bacterial identification using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
DNA was extracted from tissue or blood specimens using the QIAamp® DNA Mini Kit (QIAGEN®) following the manufacturer's recommendations, or by standard techniques of Proteinase K/SDS digestion followed by phenol chloroform extraction and ethanol precipitation, as described in [ 48].
Unlike our findings, Matsuda and co-workers reported excellent sensitivity of qPCR compared to culture for detection of these bacteria from blood specimens using these primers.
CD34+ cells, which constitute approximately 1% of nucleated blood cells in umbilical cord blood [28], were isolated from the cord blood specimen using an immunomagnetic separation technique, as previously described [35].
In this review, we examined four important differences in technique which require further investigation: the volume of blood sampled, type of blood specimen used, DNA extraction methods, and PCR primers.
DNA was isolated from blood specimens, using a modification of the Qiagen QIAmp 96 DNA Blood Kit (Qiagen GmbH, Hilden, Germany, http://www.qiagen.com).
For the SARI program a case of bacteremic pneumococcal pneumonia (BPP) was defined as the identification of S. pneumoniae in blood specimens using a single-target (lytA) quantitative real-time PCR assay adapted from Carvalho et al. [ 16]. lytA-positive specimens (cycle threshold (Ct -value < 40) were serotyped by real-time PCt -valuean adaption of the method described by Azzari et al. [ 17].
