Cobas 4800 analyzer

Cellular blood counts were delivered by the XN‐1000 analyzer (Sysmex), coagulation parameters released by STAR‐R Evolution Expert Series analyzer (Diagnostica Stago S.A.S.) and biochemical parameters from COBAS 4800 analyzer (Roche Diagnostics).

This study involved 17 patients with pulmonary adenocarcinoma from whom surgical and cytological samples were obtained from January 2015 to March 2017 at Osaka University Medical Hospital. Formalin-fixed paraffin-embedded (FFPE) tissue sections (5 µm) were prepared. DNA extraction from FFPE samples was performed according to the standard procedure of the Cobas DNA Sample Preparation kit (Roche Molecular Systems, Inc., Alameda, CA, USA). Briefly, the samples were incubated with a protease in chaotropic lysis/binding buffer to release nucleic acids and protect genomic DNA from degradation by DNase. The amount of genomic DNA was spectrophotometrically determined (Nanodrop ND-1000, Thermo Scientific, Wilmington, DE, USA) and adjusted to 2 ng/µl. DNA (150 ng) was obtained for the Cobas EGFR assay. Target DNA was amplified and detected using the Cobas 4800 Analyzer (Roche Molecular Systems, Inc.) according to the manufacturer's instructions.
The study protocol was approved by the Research Ethics Board of the Osaka University Research Committee and conducted according to Institutional Review Board guidelines (cat. no. 16293).