Assessment of a semi-automated protocol for multiplex analysis of sepsis-causing bacteria with spiked whole blood samples.

摘要: Sepsis is associated with high morbidity and mortality rates worldwide. Rapid reliable diagnostic methods are needed for efficient evidence-based treatment of septic patients. Recently, new molecular tools have emerged to complement the conventional culture-based methods. In this study, we used spiked whole blood samples evaluate together two ready-to-use solutions detection sepsis-causing bacteria. We bacterial species relevant in sepsis extracted DNA NorDiag Arrow device, using SelectNA Blood pathogen isolation kit. extracts were analyzed by polymerase chain reaction (PCR)- microarray-based Prove-it™ Bone Joint assay, resulting correctly identified limits 11–600 colony-forming unit/mL (CFU/mL). To understand recovery losses during sample preparation step capability PCR- platform respond sensitivity requirements, also determined analytical PCR microarray be 1–21 genome equivalents tested species. addition, inclusivity assay was demonstrated methicillin-resistant Staphylococcus aureus (MRSA) clones carrying SCCmec types I, II, IV, or V a nontypable type. The proof-of-concept accurate multiplex antibacterial resistance marker from selective extraction method combined high-throughput platform. Further investigations study promising potential concept sensitive, semi-automated identification pathogens directly blood.