26Purpose: The commercial PCR test SeptiFast is designed to identify DNA of individual 27 bacterial and fungal pathogens in whole blood. We aimed to evaluate the usefulness of the test 28 for detection of community onset bloodstream infections. 29 Methods:We prospectively included adult patients who were subjected to blood culture (BC) 30 at an infectious diseases department. For the evaluation one BC/PCR set (two BC bottles and 31 one PCR tube) per patient was used. When several sets were obtained and analyzed the first 32 set with any positive result was evaluated. were included in the reference standard, the PPVs for detection of these bacteria were 57%, 42 100%, 92%, 75% and 69%, respectively. 43Conclusions: Although the specificities were high, the low sensitivities and suboptimal PPVs 44 noted in the present study discourage routine use of the test in its present form for detection of 45 community onset bloodstream infections. 46 47

Background: Staphylococcus aureus bacteraemia is a disease with varying presentation, ranging from uncomplicated to life-threatening infections. In S. aureus bacteraemia, a high load of bacterial DNA in blood has been linked to mortality. We hypothesized that a high DNA load would also be linked to the presence of sepsis, and to high C-reactive protein (CRP) and lymphopaenia, indicating inflammation and immunosuppression. Methods: Twenty-seven patients with culture-proven S. aureus bacteraemia, 13 (48%) with sepsis and six (22%) nonsurvivors, were enrolled in a prospective study. Blood samples were collected on days 0, 1-2, 3-4, 6-8, 13-15 and 26-30, and subjected to droplet digital PCR targeting the nuc gene to determine the nuc DNA load. Results: nuc DNA was detected on days 0-2 in 22 patients (81%), and on days 6-8 in three patients (all non-survivors). The nuc DNA load on days 1-2 was significantly elevated in patients with sepsis (median 2.69 versus 1.32 log10 copies/mL; p ¼ .014) and in non-survivors (median 2.5 versus 1.0 log10 copies/mL; p ¼ .033). Patients with a high nuc DNA load (>3.0 log10 copies/mL) on days 1-2 had significantly elevated CRP levels at all timepoints, and significantly decreased lymphocyte counts on days 0, 1-2, 13-15 and 26-30. Conclusions: Our results indicate that a high initial load of S. aureus DNA in blood is associated with sepsis, mortality and persistent immune dysregulation in S. aureus bacteraemia patients. Further studies are needed to define the role of bacterial DNA load monitoring in the management of S. aureus bacteraemia.

We compared the performance of the BD Max enteric parasite panel to routine microscopy and an in-house PCR for the detection of Giardia intestinalis, Entamoeba histolytica, and Cryptosporidium spp. The enteric parasite panel showed good specificity for all targets and good sensitivity for E. histolytica and Cryptosporidium spp. Sensitivity for G. intestinalis with the BD Max enteric parasite panel was equivalent to that with microscopy. The World Health Organization (WHO) ranks diarrheal disease the second most common cause of morbidity and mortality in children and in the developing world (1, 2). The major etiological agents of parasitic diarrhea are considered to be Giardia intestinalis, Cryptosporidium spp., and Entamoeba histolytica (3-5).The detection of intestinal parasites can be improved, compared to microscopy, by the use of PCR-based methods (6-8). Recently, the BD Max enteric parasite panel (BD Diagnostics, Sparks, MD) was launched on the BD Max system (BD). The panel uses integrated DNA extraction and PCR to detect G. intestinalis, E. histolytica, and Cryptosporidium spp. (C. hominis and C. parvum). This study evaluated the enteric parasite panel on clinical samples and compared the performance to that of microscopy and an in-house PCR method.A total of 132 clinical samples were used for the evaluation. Overall, 39% of the samples had been stored frozen before the analyses on the BD Max system. Sixty-six samples (27 positive and 39 negative) were previously analyzed with a modified multiplexed in-house PCR for the presence of G. intestinalis, E. histolytica (9), and Cryptosporidium spp. (10) at the Department of Clinical Microbiology, Halland County Hospital, Halmstad, Sweden. Briefly, the in-house method included a prepreparation step by adding feces to lysis buffer, kept at Ϫ20°C overnight, followed by DNA extraction with a QIAsymphony DSP virus/pathogen kit (Qiagen GmbH, Germany) and a multiplex PCR run on RotorGene Q (Qiagen).The remaining samples (n ϭ 66) had previously been examined with microscopy for ova and cysts on concentrated fecal samples (32 positive and 34 negative) at the Department of Clinical Bacteriology, Sahlgrenska University Hospital, Gothenburg, Sweden. The samples analyzed with microscopy were all preserved in SAF transport medium (12.6 g/liter sodium acetate, 2% acetic acid, 4% formaldehyde, and 0.1% Triton X-100).A loop of 10 l of the fecal sample was added to the sample buffer tube (BD). The tube was vortexed and pretreated for 50 minutes on the BD prewarm heater prior to loading into the BD Max instrument along with the BD Max enteric parasite panel reagent strip. DNA extraction and real-time PCR were automatically performed by the instrument. Time from the start of sample preparation to the result, including automated data analysis, was ϳ3.5 h. The results were reported as negative or positive by the instrument. In the case of discordant results, the original sample was analyzed at a third laboratory (Ryhov County Hospital, Jönköping, Sweden) using an in-house PCR modifie...

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