Simultaneous detection of precore/basal core promoter mutations in hepatitis B virus using arrayed primer extension.

2.50
Hdl Handle:
http://hdl.handle.net/2436/15153
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.
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.
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
Submitted date:
2007-02-08
Type:
Article
Language:
en
Description:
Metadata only
ISSN:
1177-1062
Appears in Collections:
Molecular Pharmacology Research Group

Full metadata record

DC FieldValue Language
dc.contributor.authorHa, Wai-Yan-
dc.contributor.authorLau, Chi-Chiu-
dc.contributor.authorYue, Patrick Y. K.-
dc.contributor.authorHung, Kaman K. M.-
dc.contributor.authorChan, Kelvin C.-
dc.contributor.authorChui, Siu-Hon-
dc.contributor.authorChui, Albert K. K.-
dc.contributor.authorYam, Wing-Cheong-
dc.contributor.authorWong, Ricky N. S.-
dc.date.accessioned2007-12-12T10:57:57Z-
dc.date.available2007-12-12T10:57:57Z-
dc.date.issued2006-
dc.date.submitted2007-02-08-
dc.identifier.citationMolecular Diagnosis & Therapy, 10(2): 125-134en
dc.identifier.issn1177-1062-
dc.identifier.pmid16669611-
dc.identifier.urihttp://hdl.handle.net/2436/15153-
dc.descriptionMetadata onlyen
dc.description.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.en
dc.language.isoenen
dc.publisherAdisen
dc.relation.urlhttp://www.ingentaconnect.com/content/adis/mdt/2006/00000010/00000002/art00007en
dc.relation.urlhttp://direct.bl.uk/bld/PlaceOrder.do?UIN=188017433&ETOC=RN&from=searchengine-
dc.subjectHepatitis Ben
dc.subjectViral infectionsen
dc.subjectGenomicsen
dc.subjectDiagnosticsen
dc.titleSimultaneous detection of precore/basal core promoter mutations in hepatitis B virus using arrayed primer extension.en
dc.typeArticleen
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