Elecsys hbsag 2 quant reagent kits

HBsAg, anti‐HBsAg, HBeAg, and HBeAb were tested using commercial kits (Abbott Laboratory, North Chicago, IL). qHBsAg was measured using Elecsys HBsAg II Quant reagent kits (Roche Diagnostics, Indianapolis, IN). HBcAb levels were semiquantified with a chemiluminescence immunoassay (Roche Diagnostics). Serum HBV DNA levels were measured by Roche COBAS AmpliPrep/COBAS TaqMan HBV Test v2.0 (range from 20 to 1.7E+08 IU/mL; Roche Molecular Diagnostics, Branchburg, NJ). HBV genotypes and C/PreC/basal core promoter (BCP) mutations were determined by direct sequencing. Healthy controls had no history of liver or other diseases, did not have HBV infection, and had normal liver functions.

The quantitative values of the following indices were tested by Elecsys (Roche Diagnostics GmbH, Mannheim, Germany) at the noted reference ranges: HBsAb, 0 - 10 IU/L; HBeAg, <1.0 cut-off index (COI); HBeAb, >1.0 COI; and HBcAb, >1.0 COI. The HBsAg titers were quantified using Elecsys HBsAg II Quant reagent kits (Roche Diagnostics, Indianapolis, IN, USA). The detection limit of the kit was 20 IU/mL. The HBV-DNA levels were quantitated by performing real-time quantitative polymerase chain reaction (Daan GENE, Guangzhou, China). The detection limit of the assay was 100 IU/mL. The biochemical indices were detected using an autobiochemical analyzer (HITACHI 7180, Tokyo, Japan). ALT and AST were within the reference ranges of 3-35 U/L and 13-35 U/L, respectively.

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.

Blood routine indexes were assessed using an automated blood cell analyzer (Sysmex XN-3000, Kobe, Japan). Biochemical indexes of liver function were detected by an automatic biochemical analyzer (HITACHI 7600, Japan). HBeAg, HBeAb, HBcAb, and HBsAb level testing were performed using Roche Cobas E601 automatic electrochemiluminescence assay (Roche Diagnostics, IN, Germany). HBsAg was measured with Elecsys HBsAg II Quant reagent kits (Roche Diagnostics, Indianapolis, IN, USA). Serum HBV DNA was assayed with a Roche COBAS AmpliPrep/COBAS TaqMan HBV Test v2.0 (Roche Molecular Diagnostics, Branchburg, NJ, USA). These analyses were performed at the laboratory department of the third Affiliated Hospital of Sun Yat-Sen University and all procedures were performed following the manufacturer’s instructions.

Upon recruitment, patient serum was tested for HBsAb, HBeAb, HBeAg using commercial kits (Abbott Laboratories, North Chicago, IL). Quantitative HBsAg (qHBsAg) (dynamic range from 0.05 to 52,000 IU/ml) and HBsAb levels were measured with the Elecsys HBsAg II Quant reagent kits (Roche Diagnostics, Indianapolis, IN) according to the manufacturer’s instructions. Serum HBV DNA level was measured by Roche COBAS Ampliprep/COBAS TaqMan HBV Test v2.0 (dynamic range from 20 to 1.7E + 08 IU/mL, Roche Molecular Diagnostics, Branchburg, NJ). Level of fibrosis was defined by liver stiffness measurement (Fibroscan, Echosens, Paris, France). Genotyping of HBV was carried out by polymerase chain reaction-restriction fragment length polymorphism of the surface gene of HBV as previously described [11 (link), 30 (link)]. Briefly, the extracted DNA was amplified for the fragment of the HBV genome between nucleotide positions 256 and 796. The polymerase chain reaction products were subsequently treated with restriction enzymes. After incubation, the samples were run on a 3% agarose gel and stained by ethidium bromide. Six genotypes (A-F) of HBV were identified by the restriction patterns of DNA fragments. Unclassified genotype was defined as an unpredictable or atypical restriction pattern.

Upon recruitment, patient serum was tested for hepatitis B surface antibody (HBsAb), HBeAg and HBeAb, using commercial kits (Abbott Laboratories, North Chicago, IL). Quantitative hepatitis B surface antigen (qHBsAg) was measured by Elecsys HBsAg II Quant reagent kits (Roche Diagnostics, Indianapolis, IN). Serum HBV DNA levels were measured by Roche COBAS Ampliprep/COBAS TaqMan HBV Test v2.0 (dynamic range from 20 to 1.7E + 08 IU mL-1, Roche Molecular Diagnostics, Branchburg, NJ).). Six HBV genotypes (a-f) were assed by direct sequencing. The levels of fibrosis were defined by liver stiffness measurement (Fibroscan, Echosens, Paris, France).