Study on an efficient method for amplification of full-length HBV genome with relaxed-circular serum DNA
ZHANG Chunyu
JIN Weirong
DONG Hui
Abstract：ObjectiveTo establish an efficient method for amplification of full-length HBV genome with relaxed-circular serum DNA (RC-DNA).MethodsHBV RC-DNA obtained from 60 patients with chronic hepatitis B was involved in this study. First, T4 DNA polymerase and T4 DNA ligase were used to circularize the genome of HBV RC-DNA so that it became a closed circular form. Second, Phi29 DNA polymerase was applied for the rolling circle replication of closed circular RC-DNA, the product of which was then used as the template for amplification of fulllength HBV genome.ResultsWe successfully established the method for amplification of full-length HBV genome with RC-DNA, which was capable to amplify the complete HBV genome from serum samples with viral load ranging between108 to 104 copies/ml. Compared with the one-step amplification method described previously, our method was more efficient for serum samples with viral load varied from 107 to 105 copies/ml.ConclusionThe method for amplification of full-length HBV genome based on rolling circle replication of RC-DNA was efficient, and it might be a new and powerful tool for the basic and clinical study of HBV genome.
Keywords：Relaxed-circular HBV DNAFull-length genomeAmplificationRolling circle replication
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 147-150 )