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      Diagnostic performance and therapeutic impact of LightCycler SeptiFast assay in patients with suspected sepsis

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          Rapid and reliable identification of pathogens is very important in the management of septic patients. We retrospectively evaluated the diagnostic accuracy and clinical utility of a multiplex real-time polymerase chain reaction (PCR) assay (SeptiFast (SF)) in patients with suspected sepsis in a tertiary care hospital in Tallinn, Estonia. A total of 160 blood samples from 144 patients were included in the study. SF results were compared with corresponding blood culture (BC) results. The concordance between SF and BC was 78.8%. The rate of positive results was significantly higher in SF than in BC (33.7% vs. 21.2%, respectively; p < 0.001). A total of 27 samples were found positive by both SF and BC, 27 by SF only, and seven by BC only. Of a total of 83 microorganisms detected SF identified 71, and BC 42 ( p < 0.001). SF detected markedly more patients with candidemia: 11 patients were detected by SF compared to four patients by BC. Antimicrobial treatment was changed in 21 (38.9%) of 54 SF positive cases. In conclusion, our results demonstrated the high diagnostic accuracy of SF in detection of sepsis pathogens. In conjunction with its impact on therapeutic decisions, SF proved to be a useful adjunct to conventional blood culture in the diagnosis of sepsis etiology.

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          Most cited references 30

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          Impact of adequate empirical antibiotic therapy on the outcome of patients admitted to the intensive care unit with sepsis.

          Our primary goal was to evaluate the impact on in-hospital mortality rate of adequate empirical antibiotic therapy, after controlling for confounding variables, in a cohort of patients admitted to the intensive care unit (ICU) with sepsis. The impact of adequate empirical antibiotic therapy on early (<3 days), 28-day, and 60-day mortality rates also was assessed. We determined the risk factors for inadequate empirical antibiotic therapy. DESIGN Prospective cohort study. ICU of a tertiary hospital. All the patients meeting criteria for sepsis at admission to the ICU. None. Four hundred and six patients were included. Microbiological documentation of sepsis was obtained in 67% of the patients. At ICU admission, sepsis was present in 105 patients (25.9%), severe sepsis in 116 (28.6%), and septic shock in 185 (45.6%). By multivariate analysis, predictors of in-hospital mortality were Sepsis-related Organ Failure Assessment (SOFA) score at ICU admission (odds ratio [OR], 1.29; 95% confidence interval [CI], 1.19-1.40), the increase in SOFA score over the first 3 days in the ICU (OR, 1.40; 95% CI, 1.19-1.65), respiratory failure within the first 24 hrs in the ICU (OR, 3.12; 95% CI, 1.54-6.33), and inadequate empirical antimicrobial therapy in patients with "nonsurgical sepsis" (OR, 8.14; 95% CI, 1.98-33.5), whereas adequate empirical antimicrobial therapy in "surgical sepsis" (OR, 0.37; 95% CI, 0.18-0.77) and urologic sepsis (OR, 0.14; 95% CI, 0.05-0.41) was a protective factor. Regarding early mortality (<3 days), factors associated with fatality were immunosuppression (OR, 4.57; 95% CI, 1.69-13.87), chronic cardiac failure (OR, 9.83; 95% CI, 1.98-48.69) renal failure within the first 24 hrs in the unit (OR, 8.63; 95% CI, 3.31-22.46), and respiratory failure within the first 24 hrs in the ICU (OR, 12.35; 95% CI, 4.50-33.85). Fungal infection (OR, 47.32; 95% CI, 5.56-200.97) and previous antibiotic therapy within the last month (OR, 2.23; 95% CI, 1.1-5.45) were independent variables related to administration of inadequate antibiotic therapy. In patients admitted to the ICU for sepsis, the adequacy of initial empirical antimicrobial treatment is crucial in terms of outcome, although early mortality rate was unaffected by the appropriateness of empirical antibiotic therapy.
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            New developments in the diagnosis of bloodstream infections.

            New techniques have emerged for the detection of bacteria in blood, because the blood culture as gold standard is slow and insufficiently sensitive when the patient has previously received antibiotics or in the presence of fastidious organisms. DNA-based techniques, hybridisation probes, and PCR-based detection or protein-based detection by mass spectroscopy are aimed at rapid identification of bacteria and provide results within 2 h after the first signal of growth in conventional blood cultures. Also, detection of microorganisms directly in blood by pathogen-specific or broad-range PCR assays (eubacterial or panfungal) shows promising results. Interpretation is complex, however, because of detection of DNA rather than living pathogens, the risk of interfering contamination, the presence of background DNA in blood, and the lack of a gold standard. As these techniques are emerging, clinical value and cost-effectiveness have to be assessed. Nevertheless, molecular assays are expected eventually to replace the current conventional microbiological techniques for detection of bloodstream infections.
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              A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples.

              Early detection of bloodstream infections (BSI) is crucial in the clinical setting. Blood culture remains the gold standard for diagnosing BSI. Molecular diagnostic tools can contribute to a more rapid diagnosis in septic patients. Here, a multiplex real-time PCR-based assay for rapid detection of 25 clinically important pathogens directly from whole blood in <6 h is presented. Minimal analytical sensitivity was determined by hit rate analysis from 20 independent experiments. At a concentration of 3 CFU/ml a hit rate of 50% was obtained for E. aerogenes and 100% for S. marcescens, E. coli, P. mirabilis, P. aeruginosa, and A. fumigatus. The hit rate for C. glabrata was 75% at 30 CFU/ml. Comparing PCR identification results with conventional microbiology for 1,548 clinical isolates yielded an overall specificity of 98.8%. The analytical specificity in 102 healthy blood donors was 100%. Although further evaluation is warranted, our assay holds promise for more rapid pathogen identification in clinical sepsis.

                Author and article information

                European Journal of Microbiology and Immunology
                Akadémiai Kiadó, co-published with Springer Science+Business Media B.V., Formerly Kluwer Academic Publishers B.V.
                1 March 2013
                : 3
                : 1
                : 68-76
                [ 1 ] Central Laboratory, East-Tallinn Central Hospital, Tallinn, Estonia
                [ 2 ] Central Laboratory, East-Tallinn Central Hospital, Ravi 18, 10138, Tallinn, Estonia
                Author notes
                [* ] +372 6 207 493, +372 6 207 210, viive.herne@


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