Establishment of a semi-automated pathogen DNA isolation from whole blood and comparison with commercially available kits

摘要: Abstract Molecular methods for bacterial pathogen identification are gaining increased importance in routine clinical diagnostic laboratories. Achieving reliable results using DNA based technologies is strongly dependent on pre-analytical processes including isolation of target cells and their high quality purity. In this study a fast semi-automated method was established from whole blood samples compared to different commercially available kits: Looxster, MolYsis kit, SeptiFast standard EasyMAG protocol. The newly established, utilises the device combined with pre-processing steps comprising human cell lysis, centrifugation pellet resuspension. Quality assessed by universal PCR targeting 16S rRNA gene subsequent microarray hybridisation. extractions were amplified two PCR-mastermixes, allow comparison commercial mastermix guaranteed free mastermix. modified protocol Looxster kit gave most sensitive results. After hybridisation detection limit 101 102 per mL achieved depending microbial species lysed. Human present isolated suspension did not interfere lead non-specific events.