Cobas mira plus

Systolic (SBP) and diastolic blood (DBP) pressure were measured using an automatic inflation blood pressure monitor (BP3AA1-1, G-Tech, OnboElectronicCo, Schenzen, China), registered at ANVISA (No. 80275310004), following the VI Brazilian Guidelines on Hypertension [28 ].
Blood samples were collected from the antecubital vein and the serum was separated by centrifugation at 2.225 g for 15 min at room temperature (Sigma 2–3, Sigma Laborzentrifuzen, OsterodeamHarz, Germany). Blood glucose was measured using the glucose oxidase method (Cobas Mira Plus, Roche Diagnostics, GmbH, Montclair, NJ, USA), and insulin was measured by electrochemiluminescence (Modular Analytics, E170, Roche Diagnostics, GmbH, Mannheim, Germany).
Serum total cholesterol, high-density lipoprotein (HDL-C) and triglycerides were determined by an enzymatic colorimetric method (Cobas Mira Plus, Roche Diagnostics GmbH, Montclair, NJ, USA). The atherogenic index was calculated as the total cholesterol to HDL-C ratio [29 (link)].

Blood samples (5 mL) were collected by venipuncture of the external jugular vein in Ethylenediaminetetraacetic acid 10% for the complete blood count analysis and in serum for the serum biochemistry analysis; CysC determination was performed using Cobas Mira Plus Roche® based on PETIA methodology and Cistatina C Turbiquest Plus^® (Labtest, Lagoa Santa, Brazil) reagents. The sCr value was determined using an enzymatic method and a commercial kit (Kovalent, Rio de Janeiro, Brazil). The sCr concentration is considered the gold standard for characterizing normal (<1.4 mg/dL) and altered (>1.4 mg/dL) renal function in dogs. An analysis was performed to determine the behavior of sCy.
Two canine serum samples and a normal sample of canine sCy (CysC canine Escherichia coli; RD472009100) with a concentration of 0.1 mg/L were used for the validation tests. Canine CysC values were measured in canine serum samples containing low (1.48 mg/L) and high (5.69 mg/L) concentrations of this analyte, which were used to determine the precision, accuracy, quantification limit, and stability.

Blood samples were collected from patients by using a standard aseptic technique. Native blood was incubated for 60 min at room temperature; serum fractions (separated by centrifugation at 1,500 g for 15 min) were stored at −20°C until further use.
Serum high sensitivity C reactive protein (hsCRP; normal: ≤ 5 mg/l) and IgM rheumatoid factor (RF; normal: ≤ 50 IU/ml) were measured by quantitative nephelometry (Cobas Mira Plus-Roche), using CRP and RF reagents (both Dialab). ACPA (anti-CCP; aCCP) autoantibodies were detected in serum samples using a second generation Immunoscan-RA CCP2 ELISA test (Euro Diagnostica; normal: ≤ 25 IU/ml). The assay was performed according to the manufacturer's instructions.
In order to exclude the possible interference effect of RF, we compared ACE concentration and ACE2 activity values in RF positive and negative patients at baseline. We did not find statistically significant differences between positive or negative patients, thus presence of RF in the sample may not interfere with the tests (data not shown in figure).