Xe 5000 automated hematology analyzer

THB concentrations were determined from dried blood spots (DBS). Five drops of capillary blood were collected from each participant on a filter paper card (Whatman^® 903) which remained at room temperature for 4 h to dry and then stored at −80 °C. Subsequently, the samples were sent on dry ice to the University of Washington Department of Laboratory Medicine and Pathology Biomarker Laboratory for THB analysis (Seattle, WA, USA).
The THB concentrations of each participant, as measured from the DBS, were determined by a modified version of the alkaline haematin D-575 method [18 (link)]. In brief, 25 µL of eluent from a 3.2 mm diameter DBS punch in 200 µL dd/diH2O was mixed with 100 µL of THB Reagent (2.5% Triton X-100, 97.5% 0.1 M NaOH; Sigma-Aldrich, St. Louis, MO, USA), incubated at room temperature for 15 min in the dark, and the absorbance of each well was then measured at 405 nm (Synergy HT, BioTek, Winooski, VT, USA). THB of quality control DBS-matched gold-standard venous blood samples were measured on a Sysmex America XE-5000 Automated Hematology Analyzer (Mundelein, IL, USA). The THB was converted in blood-equivalent THB using a simple linear regression considering the THB of 48 quality control blood samples as gold-standard [19 (link)]. The reportable range of DBS blood-equivalent THB was 5.0–18.4 g/dL.

sCD14-ST, PCT, CRP, and WBC were determined by PATHFAST Immunoanalyzer and Reagents (fluorescent enzyme immunoassay, Mitsubishi Chemical Medience Corporation, Tokyo, Japan), Immunochramato Reader HR 201 (Immuno-chromatographic assay, B.R.A.H.M.S. GmbH, Germany), Olympus AU2700 Automated Chemistry Analyzer (Immune scatter turbidimetry, Beckman Coulter, Inc., Osaka, Japan), and Sysmex XE-5000 Automated Hematology Analyzer (Electrical impedance method and optical method, SYSMEX Corporation, Kobe, Japan), respectively. BD BACTEC FX Instrument and BD Phoenix-100 (New York) were used for blood culture and bacteria identification. Blood culture bottles and bacteria identification panels were purchased from Becton, Dickinson and Company (New York).
Collection and processing of clinical data: The name, gender, age, medical history, and blood biochemical test results of patients were recorded. Acute physiology and chronic health evaluation II (APACHE-II) scores were obtained for hematosepsis group.^{[8 ,9 (link)]} Blood specimens were taken from neonates with hematosepsis and then cultured in bacteriology room. True or false-positive results were determined in combination with clinical symptoms. This study was approved by the Ethics Committee of Longyan First Affiliated Hospital of Fujian Medical University, Longyan, China. Written informed parental consent was obtained for all patients.

The blood tests were performed using standard laboratory procedures in the Division of Laboratory Medicine, UMMC. The levels or activities of serum creatinine, alanine transaminase (ALT), aspartate aminotransferase (AST), triglycerides, total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol and fasting glucose were measured in an ADVIA^® 2400 Clinical Chemistry System (Siemens Healthcare Diagnostics, Erlangen, Germany). Sodium lauryl sulfate method was used to measure hemoglobin concentration in a XE-5000 Automated Hematology Analyzer (Sysmex Corporation, Kobe, Japan). Ion-exchange high-performance liquid chromatography assay was used for the measurement of glycated haemoglobin (HbA1c) in VARIANT™ II TURBO Hemoglobin Testing System (Bio-Rad, Hercules, CA, USA). Modified 4-variable Modification of Diet in Renal Disease study equation (Levey et al., 2006 (link)) was used to calculate the eGFR of the study participants.