Hbsag ultra

Standard ELISA procedures were followed to detect HBV seromarkers using BIORAD kits (Monolisa HBsAg ULTRA, Monolisa Anti-HBc PLUS, Monolisa Anti-HBs PLUS, BIORAD, France). DNA extraction, amplification and detection were conducted from 200 μl plasma using ABBOTT m2000sp and m2000rt System ABBOTT RealTime PCR (Abbott Molecular Inc.). The target sequence for the Abbott RealTime HBV assay is in the Surface gene in the HBV genome. This region is specific for HBV and is highly conserved. The primers are designed to hybridize to this region with the fewest possible mismatches among HBV genotypes A through H. Target region is upstream of all Tyrosine-methionine aspartate-aspartate (YMDD), HBsAg, and drug resistant mutants. The amplification cycle at which fluorescent signal is detected by the Abbott m2000rt is inversely proportional to the log of the HBV DNA concentration present in the original sample³⁹ . In order to check for consistency, 10% of the samples were retested.

All serum samples were tested for HBsAg (Monolisa™ HBsAg ULTRA), hepatitis B e antigen (HBeAg) and hepatitis B e antibody (Monolisa™ HBe Ag-Ab PLUS) using an enzyme linked immunosorbent assay (ELISA) according to the manufacturer’s instructions (Monolisa, Bio-Rad Laboratories, Marnes La Coquette).