Comparison of three different commercial PCR assays for the detection of pathogens in critically ill sepsis patients
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eiber · A. Nierhaus · S.A. Braune · G. de Heer · S. Kluge
Introduction
enable an earlier commencement of specific antibiotic treatment compared to using blood cultures. In addition, this approach may overcome the insufficiencies of initial empiric antibiotic therapies regarding multi-resistant bacteria, which are a rapidly increasing problem in ICUs. A number of different PCR-based assays are now being used increasingly to identify the causative organism in sepsis. One approach for rapid identification of the causative microorganism out of a range of potential pathogens is the so-called multiplex PCR assay (VYOO®; SIRS-Lab, Jena, Germany and LightCycler® SeptiFast; Roche, Mannheim, Germany). These assays allow for parallel detection of species- or genus-specific targets in different microorganisms. Another approach is the broad-range identification of pathogens by amplifying parts of the rRNA and other bacterial or fungal genes, followed by sequence analysis of the species (SepsiTest®; Molzym, Bremen, Germany). The vast majority of studies have investigated the LightCycler® SeptiFast assay. Furthermore, no study to date has directly compared the three different PCR assays. Therefore, the aim of this pilot study was to compare the three commercially available PCR tests with each other and with blood cultures.
Sepsis is associated with a high mortality rate and remains a serious condition in intensive care medicine [1]. Estimates for the prevalence of sepsis (or severe sepsis) and septic shock in German intensive care units (ICU) are 12 and 11%, respectively. Transposing these figures to the German population would equate to 154,000 new cases of sepsis per year. Sepsis accounts for approximately 60,000 fatalities annually and is the third most common cause of death in Germany [2]. Timely commencement of an appropriate antibiotic treatment is directly related to survival rate [3, 4, 5, 6]. Rapid and reliable detection of the causative agent is therefore crucial to optimizing treatment and improving survival rates. Blood cultures still represent the gold standard in microbiological diagnostics, alongside other cultures from localized specimens like tracheal aspirates, urine and wound swabs. Unfortunately, the currently available diagnostic tests are unable to detect the causative organism or source of infection in a considerable number of cases, despite a high clinical probability of infection [7]. Furthermore, bacteremia can only be detected in approximately 30% of patients with severe sepsis or septic shock [8]. A promising new approach to identify relevant microorganisms and their resistance genes within 6–8 h of blood sampling are assays based on the polymerase chain reaction (PCR). Following extraction and purification, nucleic acid (NA) can be rapidly amplified by PCR, and specific NAs subsequently identified. These assays should
Department of Intensive Care Medicine, University Medical Center Hamburg-Eppendorf, Hamburg
Comparison of three different commercial PCR assays for the detection of pathogens in criti
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