Study population and criteria for inclusion and exclusionAll study participants were 18 years or older, had a body mass index < 40, and provided written informed consent. Patients were recruited as being likely to enter the Sepsis cohort if they met the ACCP/SCCM Consensus Statement [6], and had a clinical selleckchem Pazopanib suspicion of systemic infection based on microbiological diagnoses. A definitive diagnosis of sepsis was unlikely to be known at the time patients were enrolled in the study; thus confirmation of sepsis and assignment of patients to the sepsis cohort was made retrospectively.Potential sepsis participants admitted to the ICU and, patients admitted for planned major open surgery, were excluded from the study if they had any systemic immunological disorders including Systemic Lupus Erythromatosus, Crohn’s disease and Insulin-Dependent Diabetes Mellitus (Type 1 diabetes); were transplant recipients or were currently receiving chemotherapy treatment for cancer.

Twenty-seven blood culture positive sepsis patients with community-acquired infections were enrolled into this clinical trial as soon as practicable, on admission to the ICU. Specifically, sepsis patients’ were enrolled within 24 hours of admission and on average, were in the ICU for five days. Participants in the post-surgical (PS) cohort were recruited pre-operatively and blood samples were collected within 24 hours following surgery (n = 38). Furthermore, 20 healthy adult control (HC) participants were recruited within the Mater Adult Hospital staff, on the basis that they had no concurrent illnesses at the time of blood collection or any past history of immunological dysfunction.

All participants or their surrogate decision-maker provided written informed consent prior to the collection of any study data or biological samples.Collection of dataDemography, vital signs measurements (blood pressure, heart rate, respiratory rate, tympanic temperature), haematology (full blood count), clinical chemistry (urea, electrolytes, liver function enzymes, blood glucose) as well as microbial status were recorded. Blood was drawn into minimally two PAXgene (PreAnalytix, Feldbachstrasse, Hombrechtikon, Switzerland) tubes (5 ml total) for gene expression analyses using the SeptiCyte Lab test.Gene expression assaysRNA isolation was performed using PAXgene Blood RNA kits (PreAnalytix, a Qiagen/BD Company, Feldbachstrasse, Switzerland) and following standard instructions recommended by the manufacturer.

RNA quality was determined using an automated electrophoresis station (Experion, BioRad Batimastat (Gladesville, New South Wales, Australia) and BioAnalyser, Agilent (Forest Hill, Victoria, Australia). The 260 nm/280 nm ratios for all samples were > 1.9. Once total RNA was extracted, gene expression was assessed using the Affymetrix HGU133 Plus 2.