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Rapid and sensitive method for the detection of serum hepatitis B virus DNA using the polymerase chain reaction
S Kaneko1, S M Feinstone, R H Miller
1Hepatitis Viruses Section, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
Abstract:
We have developed a rapid procedure for the detection of serum hepatitis B virus (HBV) DNA using the polymerase chain reaction (PCR) technique. HBV DNA is released from virions by incubating serum with 0.1 M NaOH for 60 min at 37 degrees C. The mixture is brought to neutral pH with HCl, and the HBV DNA sequences are detected by agarose gel electrophoresis and ethidium bromide staining after PCR amplification with two successive sets of primer pairs. The detection limit of this method (i.e., 10(-5) pg of HBV DNA) is equivalent to that previously determined by one round of PCR amplification and Southern blot hybridization analysis. The advantages are that the assay can be completed in 1 day, is very sensitive, and does not require the use of radiolabeled reagents.