title: Bacterial and fungal DNA extraction from blood samples: automated protocols.
abstract: Automation in DNA isolation is a necessity for routine practice employing molecular diagnosis of infectious agents. To this end, the development of automated systems for the molecular diagnosis of microorganisms directly in blood samples is at its beginning. Important characteristics of systems demanded for routine use include high recovery of microbial DNA, DNA-free containment for the reduction of DNA contamination from exogenous sources, DNA-free reagents and consumables, ideally a walkaway system, and economical pricing of the equipment and consumables. Such full automation of DNA extraction evaluated and in use for sepsis diagnostics is yet not available. Here, we present protocols for the semiautomated isolation of microbial DNA from blood culture and low- and high-volume blood samples. The protocols include a manual pretreatment step followed by automated extraction and purification of microbial DNA.
authors.affiliation: Molzym GmbH & Co. KG, Mary-Astell-Straße 10, 28359, Bremen, Germany, lorenz@molzym.com.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Lorenz
authors.forename: Claudia
authors.initials: C
authors.lastname: Disqué
authors.forename: Helge
authors.initials: H
authors.lastname: Mühl
mesh_terms: D057205:Automation, Laboratory / Q000295:instrumentation / Q000379:methods*; D016470:Bacteremia / Q000175:diagnosis* / Q000382:microbiology / Q000473:pathology; D001419:Bacteria / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D001800:Blood Specimen Collection; D003470:Culture Media / Q000737:chemistry; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D016469:Fungemia / Q000175:diagnosis* / Q000382:microbiology / Q000473:pathology; D005658:Fungi / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000295:instrumentation / Q000379:methods
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial; D004271:DNA, Fungal