Detection of hepatitis B virus sequences in serum by using in vitro enzymatic amplification

摘要: Abstract In vitro enzymatic amplification was applied to detect hepatitis B virus (HBV) DNA sequences in serum. This technique, known as the polymerase chain reaction (PCR) used amplify a 128 bp fragment including 112 nucleotide long sequence complementary region S gene of HBV genome. Amplified samples were subjected spot-test hybridization and scintillation counting using 32P-labeled oligonucleotide probe. A kinetic study, performed for 4 32 PCR cycles with viral particle preparation, showed time-limited exponential accumulation specific amplified fragment. Amplification yield after at least × 106 detection limit equal 3 102 particles per ml As reliability technique greatest 24 cycles, these conditions develop quantitative test 104 Results this perfectly correlated those obtained from classical spot without amplification. Ethydium bromide stained agarose gel Southern blot analysis confirmed