E170 analyzer

CEA and CYFRA 21-1 levels in the CSF and serum were measured with an immunochemiluminescent assay on a Roche MODULAR ANALYTICS E170 analyzer. Specifically, the serum or CSF sample, biotinylated monoclonal CEA-specific or CYFRA 21-1-specific antibody, and a monoclonal CEA-specific or CYFRA 21-1-specific antibody (Elecsys CEA and CYFRA 21-1 immunochemiluminescent assay kit) labeled with a ruthenium complex were reacted to form a sandwich complex. After addition of streptavidin-coated microparticles, the complex bound to the solid phase via the interaction of biotin and streptavidin. The microparticles were then magnetically captured onto the surface of the electrode, and unbound substances were removed. Application of voltage to the electrode then induced chemiluminescent emission, which was measured by a photomultiplier. The results were determined via reference to a calibration curve that was instrument-specifically generated. The lower detection limits for CEA and CYFRA 21-1 were 0.2 μg/mL and 0.1 ng/mL, respectively. Measurement of a panel of TMs, including CEA and CYFRA 21-1, is a routine aspect of the CSF analysis workup for ascertaining the diagnosis of intracranial malignant metastasis in our institution and had been approved by the institutional ethics committee.

The serum specimens were collected and assayed for HBsAg and HBeAg at 1, 4, 7 and 10 dpi. The levels of HBsAg, HBsAb, HBcAb and HBeAg in serum were detected by using either a commercial ELISA kit (Kehua, Shanghai, China) or an electrochemiluminescence immunoassay (ECLIA) on a modular analytics E170 analyzer (Roche, Indianapolis, IN, USA) according to the manufacturer’s instructions. 10-fold diluted serum samples were used for detection. Serum HBV DNA was extracted using a QIAamp MinElute Virus Spin kit (Qiagen, Hilden, Germany) and was quantitatively detected by real-time PCR (qPCR) using the SYBR green qPCR master mix (Qiagen, Hilden, Germany). Six mice were included per group. Melt curve analysis and agarose gel electrophoresis were used to verify the specificity of the qPCR. The following primers were used: forward primer: 5′-CTG CAT CCT GCT GCT ATG-3′ (nt 408–425), and reverse primer: 5′-CAC TGA ACA AAT GGC AC-3′ (nt 685–701) according to the reference sequence with GenBank accession number (AY220698.1). A serum sample containing a known concentration of HBV DNA was used as positive control.

Eighty-six patients received serum creatine kinase (CK) measurements. Thirty-two and forty-four of them further received measurements of serum aspartate aminotransferase (AST) and lactic dehydrogenase (LDH) respectively. Twenty-nine patients received blood lipid measurements (including blood total cholesterol (TC), blood low-density lipoprotein (LDL) and blood triglyceride (TG). The measurements of hormone level were performed with Roche cobas hormonal assay on Roche E170 analyzer. The values of CK, AST, LDH, lipids and hormone level were obtained before medication intervention. All the laboratory tests were performed by the clinical laboratory. Reference range of each test was reported according to the lab normative values of Huashan Hospital.