A rapid real-time quantitative polymerase chain reaction for hepatitis B virus

Quantification of hepatitis B virus (HBV) DNA in serum is important for mon itoring treatment. A rapid and cost effective alternative to the methods av ailable currently was developed based on a real-time quantitative polymeras e chain reaction (PCR) done in the LightCycler(TM) apparatus. Primers and a probe For sequences of the surface gene of HBV were designed and quantific ation 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 fr om 4 x 10(2) to 1.3 x 10(10) surface gene copies/ml serum. Nested PCR was r equired For quantification in the lower part of this range ( < 10(5) copies ). The real-time PCR and Amplicor Monitor (Roche) tests performed comparabl y at virus concentrations below 10(6) copies/ml. The commercial test undere stimated higher concentrations of virus. (C) 2001 Elsevier Science B.V. All rights reserved.