Architect hbsag reagent kit

Total DNA was extracted with a QIAamp DNA Blood Mini Kit (Qiagen) from 100 μL of the culture supernatant or 1.0 × 10⁷ cells, and 5 μL of the extracted DNA solution was subjected to RT‐PCR using a LightCycler 480 system (Roche Diagnostics) with SYBR Green I Master (Roche Diagnostics), primers HBXF11 (5′‐ATG GCT GCT AGG CTG TGC TG‐3′) and HBXR4 (5′‐GTC CGC GTA AAG AGA GGT GC‐3′). Total RNA was extracted from 1.0 × 10⁷ cells using Trizol (Invitrogen) and subjected to cDNA synthesis by SuperScript III (Invitrogen). RT‐PCR for quantifying total amount of pregenomic RNA and precore messenger RNA (mRNA) was conducted with SYBR Green I Master, primers HBCF7 (5′‐GCT ACC TGG GTG GGT GGT AAT TTG G‐3′) and HBCR5 (5′‐AGG GGA CCT GCC TCG TCG TCT AAC‐3′). The HBsAg and hepatitis B e antigen (HBeAg) levels in the culture supernatant were quantified with a chemiluminescence immunoassay (CLIA) using the ARCHITECT HBsAg reagent kit and ARCHITECT HBeAg reagent kit (Abbott Laboratories, Chicago, IL), respectively.

Venous blood samples were collected from infants at the age of 7 months (1 month after the last dose of the vaccine). The detection of HBsAg in infants was carried out through the chemiluminescent microparticle immunoassay (CMIA) with an Abbott ARCHITECT HBsAg Reagent Kit (Abbott Laboratories, North Chicago, Illinois, USA). An ARCHITECT anti-HBs Reagent Kit (Abbott Laboratories) from CMIA was used to determine the anti-HBs levels, with a range of 0–15000 mIU/mL after 15 dilutions. Infants with serum anti-HBs <100 mIU/mL and anti-HBs ≥100 mIU/mL at 7 months of age were classified as low responders (LRs) and high responders (HRs), respectively. Maternal HBV DNA was assessed by the Roche TaqMan HBV test (Roche Diagnostics, Grenzach, Germany).

The following were measured from blood serum samples collected before and at the end of DAA therapy: levels of HBsAg and anti-HBs, hepatitis B core antibodies (anti-HBc), and HBV-DNA. Methods used for measurements were as follows: chemiluminescent immunoassay (CLIA) for HBs antigens (ARCHITECT HBsAg Reagent Kit; Abbott, Chicago, IL); CLIA for HBs antibodies (ARCHITECT Anti-HBs Reagent Kit; Abbott); CLIA for HBc antibodies (ARCHITECT Anti-HBc II Reagent Kit; Abbott), and the TaqMan probe for HBV-DNA (COBAS Ampriprep/COBAS TaqMan HBV Test, v2.0; Roche, Basel, Switzerland).