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Quantification of hepatitis B virus (HBV) DNA in serum is important for monitoring treatment. A rapid and cost effective alternative to the methods available currently was developed based on a real-time quantitative polymerase chain reaction (PCR) done in the LightCycler apparatus. Primers and a probe for sequences of the surface gene of HBV were designed and quantification achieved by reference to standards containing known concentrations of the target sequence. A single copy of the HBV genome could be detected if present in the reaction mixture. The quantitative range of the assay was from 4x10 2 to 1.3x10 10 surface gene copies/ml serum. Nested PCR was required for quantification in the lower part of this range (<10 5 copies). The real-time PCR and Amplicor Monitor (Roche) tests performed comparably at virus concentrations below 10 6 copies/ml. The commercial test underestimated higher concentrations of virus.
Molecular Biology Unit, Sexually Transmitted and Blood Borne Virus Laboratory, Central Public Health Laboratory, 61 Colindale Avenue, London NW9 5HT, UK
Molecular Biology Unit, Sexually Transmitted and Blood Borne Virus Laboratory, Central Public Health Laboratory, 61 Colindale Avenue, London NW9 5HT, UK