Cobas taqman hbv test v2

All routine laboratory data were collected immediately before the first NA treatment and serially during the treatment. Serum M2BPGi level was measured using serum samples that had been stored at −20 °C. M2BPGi quantification was performed by an immunoassay using a commercially available kit (HISCL M2BPGi; Sysmex Co., Kobe, Japan) and a fully automatic immunoanalyzer (HISCL-5000; Sysmex Co.). Serum HBV viral load was determined using a COBAS TaqMan HBV test v2.0 (Roche Diagnostics, Branchburg, NJ), which has a dynamic range of 2.1–9.0 log copies (LC)/mL. Quantitative measurements of HBsAg, Hepatitis B e-antigen (HBeAg), and hepatitis B core-related antigen (HBcrAg) were conducted using commercial chemiluminescent enzyme immunoassay kits.

Blood was collected from the 51 subjects to measure biochemical values at week 0 (before the experiment), and the 4th, 12th, 24th, 48th weeks (in the experiment) and in the 4th and 12th weeks after the experiment. Heparinized blood was collected to evaluate liver and kidney function and genotype, and to quantify HBV-DNA concentration, HBsAg, and HBeAg. Serum HBV-DNA levels (copies/mL) were measured by polymerase chain reaction (PCR) using Cobas TaqMan HBV Test, v2.0 (Roche Molecular Systems, Pleasanton, CA, USA), according to the manufacturer’s instructions. Quantification of HBsAg was performed by an automated chemiluminescent micro-particle immunoassay (CMIA) using the Roche Cobas e602 analyzer with Elecsys HBsAg II Quant reagent kits (Roche Diagnostics, Santa Clara, CA, USA), according to the manufacturer’s instructions. Quantification of HBeAg was conducted by an automated chemiluminescent micro-particle immunoassay (CMIA) using the Roche Cobas e602 analyzer with Elecsys HBeAg Quant reagent kits (Roche Diagnostics, CA, USA), according to the manufacturer’s instructions. The remaining blood was segregated into serum and cells. Serum was stored as aliquots in liquid nitrogen until analysis and cells were analyzed to determine the T cell subtypes.

The biochemical, serological, and virological parameters were measured using standard laboratory procedures. HBV serology included HBsAg, anti-HBs, hepatitis B e-antigen (HBeAg), anti-HBe, and anti-HBc tests. Serum HBsAg test was performed using commercial kits (Abbott Laboratories; Lake Bluff, IL, USA). Serum HBV DNA levels were assessed via real-time polymerase chain reactions (COBAS TaqMan HBV Test v2.0, Roche Diagnostics, Branchburg, NJ, USA). A serum HBV DNA level at 20 IU/mL was set as the threshold for HBV-DNA positivity. HBeAg seroconversion was defined by the disappearance of HBeAg with or without detectable anti-HBe.

Nineteen HCC patients who underwent surgical resection at Kobe University Hospital were enrolled in this study. Of the 19 patients, 9 (age 69.7 ± 5.6 years, 8 males and 1 female) were serologically negative for HBsAg, while 10 (age 53.0 ± 11.5 years, 9 males and 1 female) were positive for HBsAg. Laboratory data including platelet counts, transaminase levels, haemoglobin A1c, hyaluronic acid and quantitative HBsAg levels (HBsAg-HQ: Fujirebio, Tokyo, Japan) at the time of surgery were retrieved from patients’ medical records. In addition, hepatitis B core antibody (anti-HBc; Architect AUSAB, Abbott Japan, Tokyo, Japan) and HBV DNA levels (Cobas® TaqMan HBV Test, v2.0, Roche Diagnostics, Basel, Switzerland) were also examined. The cut-off levels of HBsAg-HQ and HBV DNA were estimated 5 mIU/ml and 2.06 log copies/ml, respectively. Patients negative for anti-HBc or positive for hepatitis C virus antibodies were excluded from this study.

HBsAg was measured using the Architect HBsAg QT assay (Abbott Japan, Tokyo), and anti-HCV antibodies were measured using the Architect HCV assay (Abbott Japan, Tokyo). HBV-DNA was measured using COBAS TaqMan HBV Test v. 2.0 (Roche Diagnostics K.K., Tokyo, Japan), and HCV-RNA was measured using the COBAS TaqMan HCV Test v. 2.0 (Roche Diagnostics K.K.).