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Wolverhampton Intellectual Repository and E-Theses > Research Institutes > Research Institute in Healthcare Science > Molecular Pharmacology Research Group > Simultaneous detection of precore/basal core promoter mutations in hepatitis B virus using arrayed primer extension.

Please use this identifier to cite or link to this item: http://hdl.handle.net/2436/15153
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Title: Simultaneous detection of precore/basal core promoter mutations in hepatitis B virus using arrayed primer extension.
Authors: Ha, Wai-Yan
Lau, Chi-Chiu
Yue, Patrick Y. K.
Hung, Kaman K. M.
Chan, Kelvin C.
Chui, Siu-Hon
Chui, Albert K. K.
Yam, Wing-Cheong
Wong, Ricky N. S.
Citation: Molecular Diagnosis & Therapy, 10(2): 125-134
Publisher: Adis
Issue Date: 2006
URI: http://hdl.handle.net/2436/15153
PubMed ID: 16669611
Additional Links: http://www.ingentaconnect.com/content/adis/mdt/2006/00000010/00000002/art00007
http://direct.bl.uk/bld/PlaceOrder.do?UIN=188017433&ETOC=RN&from=searchengine
Abstract: BACKGROUND: 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.
Type: Article
Language: en
Description: Metadata only
Keywords: Hepatitis B
Viral infections
Genomics
Diagnostics
ISSN: 1177-1062
Appears in Collections: Molecular Pharmacology Research Group

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