Simultaneous detection of precore/basal core promoter mutations in hepatitis B virus using arrayed primer extension.
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Yue, Patrick Y. K.
Hung, Kaman K. M.
Chan, Kelvin C.
Chui, Albert K. K.
Wong, Ricky N. S.
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AbstractBACKGROUND: Hepatitis B is a major disease that causes serious public health problems worldwide. The loss of HBeAg expression due to point mutations or single nucleotide polymorphisms (SNPs) in the precore/basal core promoter region of the hepatitis B virus (HBV) is associated with hepatocellular cirrhosis and carcinoma. Simultaneous screening for these mutations is strongly advocated for monitoring disease development in HBV-infected patients. The aim of this study is to apply arrayed primer extension (APEX) for the detection of HBV SNPs at the precore/basal core promoter. METHODS AND RESULTS: We optimized APEX for simultaneous detection of eight potential sites of SNPs in the precore/basal core promoter region of HBV. The precore/basal core promoter regions of HBV from 36 HBV-infected patients were amplified by PCR. After purification and DNA fragmentation, the short, single-stranded HBV DNA fragments were allowed to hybridize with the oligonucleotides corresponding to the sites of SNPs immobilized on glass slides, followed by incorporation of different fluorescently labeled dideoxynucleotides. This allows fast and unequivocal discrimination between wild-type and mutant genotypes with high dideoxy-nucleotide incorporation efficiency, sensitivity, and specificity. The coexistence of both genotypes was also detected; this was undetected by DNA sequencing. CONCLUSION: The simultaneous detection of SNPs in HBV precore/basal core promoter by APEX enables large-scale diagnostic analysis, which can be extended to the whole HBV genome.
CitationMolecular Diagnosis & Therapy, 10(2): 125-134
CollectionsMolecular Pharmacology Research Group
- Different hepatitis B virus genotypes are associated with different mutations in the core promoter and precore regions during hepatitis B e antigen seroconversion.
- Authors: Chan HL, Hussain M, Lok AS
- Issue date: 1999 Mar
- Hepatitis B genotypes and precore/basal core promoter mutants in HBeAg-negative chronic hepatitis B.
- Authors: Lin CL, Liao LY, Liu CJ, Chen PJ, Lai MY, Kao JH, Chen DS
- Issue date: 2002
- [The prevalence and clinical significance of precore and core promoter mutations in Korean patients with chronic hepatitis B virus infection].
- Authors: Kim HJ, Yoo BC
- Issue date: 2002 Jun
- Hepatitis B virus: prevalence of precore/core promoter mutants in different clinical categories of Indian patients.
- Authors: Gandhe SS, Chadha MS, Walimbe AM, Arankalle VA
- Issue date: 2003 Sep
- Analysis of the precore and core promoter DNA sequence in liver tissues from patients with hepatocellular carcinoma.
- Authors: Cho SW, Shin YJ, Hahm KB, Jin JH, Kim YS, Kim JH, Kim HJ
- Issue date: 1999 Aug