Automation of Septifast？ for molecular diagnosis of infection in septic patients

Inclusion criteria were age ≥18 years and suspected sepsis. The Ethics Committees from our institution approved the study, and patient or relatives' consent was obtained for blood sampling for PCR. Patient population (ICU): 100 Septifast？ determinations and correlative blood cultures and microbiological cultures were performed in 64 patients with systemic inflammatory response syndrome and high clinical risk factors for bloodstream infection.We used a new multiplex PCR-based test, LightCycler？ SeptiFast？ Test MGRADE (Roche Diagnostics, Penzberg, Germany), including software for simultaneous detection of sepsis-relevant microorganisms directly from blood, according to the manufacturer's recommendations. We followed the standard manufacturer's recommended procedure for extraction and compared it with the procedure using the MagNa pure？ compact (Roche Applied Science, Penzberg, Germany) extraction column based on magnetic nanoparticles. Samples for blood culture analysis were drawn from a fresh venipuncture site according to the common guidelines (DGHM). Blood cultures were analyzed using a semi-automated blood system (BacT/ALERT？; bioMerieux, Marcy-Etoile, France). VITEK II (bioMerieux) was the system used to phenotypically identify pathogens growing from blood cultures and other microbiological tests (stool, urine, respiratory samples, catheter, etc.).The results were both retrospective and observational. The quality of PCR results versus other laboratory findings and clinical evaluation, using sensitivity and specificity analysis and kappa evaluation to compare the results obtained, was determined. One hundred determinations were studied comparing both methods for extraction. Table 1 presents comparative data for both methods. For sensitivity and specificity analysis, the blood culture data define the disease condition (infection).MagNa pure？ extraction versus the standard procedure for extraction used in the SeptiFast？ test reduces the total test time from 6 to 3 o