We enrolled all patients

with chronic hepatitis B (CHB) at Severance Hospital (Seoul, South Korea) or CHA Bundang Medical Center (Seongnam-Si, South Korea) who were started on ETV (0.5 mg once a day) between January 2007 and June 2008 and for whom stored serum was available. The inclusion criteria were the presence of serum HBsAg for 6 or more months, HBV genotype C, an age greater than 16 years, and a Afatinib mouse previous lack of treatment with a subsequent ETV treatment period of at least 24 months. ETV was commenced when the HBV DNA level was more than 10,000 copies/mL and when either the alanine aminotransferase (ALT) level was greater than 2 times the upper limit of normal or biopsy showed significant fibrosis/cirrhosis.25 The exclusion criteria were a coinfection with hepatitis C virus

or human immunodeficiency virus, a history of organ transplantation, decompensated liver cirrhosis (ascites, varices, encephalopathy, albumin level < 3 mg/dL, total bilirubin level > 2.5 mg/dL, or prothrombin time > 3 seconds longer than normal), and a concurrent use of immunomodulatory drugs or corticosteroids. Written, informed consent was obtained from all participating patients. This study Selleckchem Metformin was approved by the local institutional review board and was conducted in accordance with the principles set forth in the Declaration of Helsinki. Routine biochemical tests, including ALT, albumin, total bilirubin, and creatinine levels, were performed with a sequential multiple autoanalyzer. The Architect HBsAg QT immunoassay (Abbott Diagnostic, very Wiesbaden, Germany) was used to quantify qHBsAg according to the manufacturer’s instructions.5, 13

Briefly, the assay was carried out in two steps: HBsAg present in the sample was bound to antibody to hepatitis B surface antigen (anti-HBs)–coated microparticles, and an acridinium-labeled anti-HBs conjugate was added together with pretrigger and trigger solutions. The products of the resulting chemiluminescent reaction were measured in relative light units. The qHBsAg calibration curve ranged from 0.05 to 250 IU/mL, and the samples were diluted with a diluent (1:20 or 1:250) as needed to expand the detection range. The Architect platform (Abbott Diagnostic) was also used to quantify qHBeAg. Briefly, qHBeAg was measured with an automated microparticle chemiluminescent immunoassay based on a previously described method.