Cobas c702 analyzer

Blood samples for the measurement of routine clinical biochemistry parameters were collected in EDTA-lined vacuum tubes and in gel tubes containing lithium heparin. Plasma concentrations of total cholesterol, low-density lipoprotein (LDL) cholesterol (LDL-C), high-density lipoprotein (HDL)-C, and triglycerides were measured at the Department of Medical Biochemistry (Oslo University Hospital Rikshospitalet, Oslo, Norway) by colorimetric and/or enzymatic methods on a Cobas c702 analyzer (Roche Diagnostics International Ltd., Rotkreuz, Switzerland).

All expressed biomarkers were measured in the serum of patients’ blood. All samples were obtained by venipuncture into serum monovettes^® and centrifuged at 2000 g for 10 min at 20 °C. The aliquoted samples were cooled down with liquid nitrogen before being stored at − 80 °C until analysis. The complete processing was conducted within two hours after blood extraction. After thawing, the samples were mixed gently by inverting and centrifuged with 2500g for 10 min at 20 °C, respectively, 3000g for 30 min for hsTnI at 4 °C.
HsTnT was measured with the Troponin T hs STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The limit of blank (LoB) for this assay was 3 ng/L and the limit of detection (LoD) was 5 ng/L as described in the instructions for use [15 ]. HsTnI was measured with the STAT High sensitivity Troponin-I assay on an Architect i1000 analyzer (Abbott, Wiesbaden, Germany). The LoB was 0.7–1.3 ng/L and the LoD was 1.1–1.9 ng/L for this assay as described in the instructions for use [16 ]. NT-proBNP was measured with the proBNP II STAT assay on a cobas e 602 analyzer (Roche Diagnostics, Mannheim, Germany). The LoD for this assay was 5 ng/L [17 ]. Creatinine was measured with the Creatinine Jaffe Gen.2 assay on a cobas c 702 analyzer (Roche Diagnostics, Mannheim, Germany).

Serum AST and ALT concentrations were measured using the method recommended by the International Federation of Clinical Chemistry (IFCC) with pyridoxal phosphate activation at 37 °C with the Cobas c 702 Analyzer (Roche, Basel, Switzerland).
The normal range of values for AST in serum was 10–33 U/L in women and 10–35 U/L in men. In comparison, the normal range of values for ALT in serum was 6–41 U/L in women and 9–59 U/L in men. The AST to ALT ratio (AST/ALT ratio) is the ratio between the concentrations of AST and ALT enzymes.

Pleural samples were analysed for total differential cell counts, CRP, ADA, glucose, total protein, albumin, lactate dehydrogenase (LDH), pH. For all pleural specimens cytologic examination and bacterial cultures were obtained. Furthermore, samples were examined for mycobacteria using Ziehl-Neelsen stain and culture.
The supernatant of each sample was obtained by centrifugation at 300 rpm for 15 min and stored at 20 °C until being assayed for the CRP measurement. CRP measurements were performed by particle-enhanced immunoturbidimetric assay with the cobas c 702 analyzer, using the Tina-quant C-Reactive protein IV kit (Roche Diagnostics, Mannheim, Germany). The appropriate control was provided by the same company and assays were performed according to the manufacturer’s instructions by the Department of Microbiology, University Hospital of Larissa, Larissa, Greece. The measuring range was 0.6–350mg/dl.
Total pleural fluid ADA was determined by the Giusti method which is based on the measurement of ammonia released from adenosine when converted to inosine. Pleural fluid was centrifuged and the supernatant was incubated in adenosine buffer at 37 °C, followed by incubation with Berthelot reagent at 37 °C and subsequent photometric analysis at 405 nm using a Secomam Basic semi- automatic analyser.

Complete blood cell counts, including erythrocyte, platelet, and total and differential leukocyte (neutrophil, lymphocyte, monocyte, eosinophil, and basophil) counts were assessed using an automated hematology analyzer (Coulter Counter STKS, Beckman Coulter, Fullerton, CA). Routine chemistry tests, including liver and kidney function tests were performed using a Cobas c702 analyzer (Roche Diagnostics, Basel, Switzerland).