title: Detection of hepatitis B virus DNA by real-time PCR using TaqMan-MGB probe technology.
abstract: OBJECTIVE
  To develop a real-time PCR for detecting hepatitis B virus (HBV) DNA based on TaqMan technology using a new MGB probe.
  
  
  METHODS
  Plasmid containing the sequence of X gene (1414-1744 nt) was constructed as HBV-DNA standard for quantitative analysis. A TaqMan-MGB probe between primers for amplification was designed to detect PCR products. The interested sequence contained in the plasmid and in clinical specimens was quantitatively measured.
  
  
  RESULTS
  The detection limit of the assay for HBV DNA was 1 genome equivalent per reaction. A linear standard curve was obtained between 10(0) and 10(9) DNA copies/reaction (r>0.990). None of the negative control samples showed false-positive reactions in duplicate. HBV DNA was detected in 100% (50/50) of HBV patients with HbeAg, and in 72.0% (36/50) with HBsAg, HBeAb and HBcAb. The coefficient of variation for both intra- and inter-experimental variability demonstrated high reproducibility and accuracy.
  
  
  CONCLUSIONS
  Real-time PCR based on TaqMan-MGB probe technology is an excellent method for detection of HBV DNA.
authors.affiliation: Institute of Genetic Diagnosis, Fourth Military Medical University, 17 Changle West Road, Xi'an 710032, Shaanxi Province, China. zhaojrr@fmmu.edu.cn
authors.forename: Jin-Rong
authors.initials: JR
authors.lastname: Zhao
authors.forename: Yu-Jie
authors.initials: YJ
authors.lastname: Bai
authors.forename: Qing-Hua
authors.initials: QH
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Wan
authors.forename: Ding
authors.initials: D
authors.lastname: Li
authors.forename: Xiao-Jun
authors.initials: XJ
authors.lastname: Yan
mesh_terms: D001483:Base Sequence; D015342:DNA Probes / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis; D006515:Hepatitis B virus / Q000235:genetics* / Q000302:isolation & purification*; D019694:Hepatitis B, Chronic / Q000175:diagnosis*; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D011994:Recombinant Proteins / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019914:Taq Polymerase
chemical_list: D015342:DNA Probes; D004279:DNA, Viral; D011994:Recombinant Proteins; D019914:Taq Polymerase