Study of hepatitis B virus gene mutations with enzymatic colorimetry-based DNA microarray

  • Mao H
  • Wang H
  • Zhang D
 et al. 
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OBJECTIVES: To establish a modified microarray method for detecting HBV gene mutations in the clinic. DESIGN AND METHODS: Site-specific oligonucleotide probes were immobilized to microarray slides and hybridized to biotin-labeled HBV gene fragments amplified from two-step PCR. Hybridized targets were transferred to nitrocellulose membranes, followed by intensity measurement using BCIP/NBT colorimetry. RESULTS: HBV genes from 99 Hepatitis B patients and 40 healthy blood donors were analyzed. Mutation frequencies of HBV pre-core/core and basic core promoter (BCP) regions were found to be significantly higher in the patient group (42%, 40% versus 2.5%, 5%, P < 0.01). Compared with a traditional fluorescence method, the colorimetry method exhibited the same level of sensitivity and reproducibility. CONCLUSIONS: An enzymatic colorimetry-based DNA microarray assay was successfully established to monitor HBV mutations. Pre-core/core and BCP mutations of HBV genes could be major causes of HBV infection in HBeAg-negative patients and could also be relevant to chronicity and aggravation of hepatitis B.

Author-supplied keywords

  • *Colorimetry
  • *Oligonucleotide Array Sequence Analysis
  • Electrophoresis
  • Hepatitis B virus/*genetics
  • Hepatitis B/virology
  • Humans
  • Indoles
  • Mutation
  • Nitroblue Tetrazolium
  • Polymerase Chain Reaction
  • Reproducibility of Results
  • Sequence Analysis, DNA

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  • H Mao

  • H Wang

  • D Zhang

  • J Zhao

  • J Shi

  • Z Cui

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