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

TitleSimultaneous detection of precore/basal core promoter mutations in hepatitis B virus using arrayed primer extension
Authors
Issue Date2006
PublisherAdis International Ltd. The Journal's web site is located at http://moleculardiagnosistherapy.adisonline.com/
Citation
Molecular Diagnosis And Therapy, 2006, v. 10 n. 2, p. 125-134 How to Cite?
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 dideoxynucleotide 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. © 2006 Adis Data Information BV. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/78870
ISSN
2015 Impact Factor: 2.602
2015 SCImago Journal Rankings: 0.944
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorHa, WYen_HK
dc.contributor.authorLau, CCen_HK
dc.contributor.authorYue, PYKen_HK
dc.contributor.authorHung, KKMen_HK
dc.contributor.authorChan, Ken_HK
dc.contributor.authorChui, SHen_HK
dc.contributor.authorChui, AKKen_HK
dc.contributor.authorYam, WCen_HK
dc.contributor.authorWong, RNSen_HK
dc.date.accessioned2010-09-06T07:47:49Z-
dc.date.available2010-09-06T07:47:49Z-
dc.date.issued2006en_HK
dc.identifier.citationMolecular Diagnosis And Therapy, 2006, v. 10 n. 2, p. 125-134en_HK
dc.identifier.issn1177-1062en_HK
dc.identifier.urihttp://hdl.handle.net/10722/78870-
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 dideoxynucleotide 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. © 2006 Adis Data Information BV. All rights reserved.en_HK
dc.languageengen_HK
dc.publisherAdis International Ltd. The Journal's web site is located at http://moleculardiagnosistherapy.adisonline.com/en_HK
dc.relation.ispartofMolecular Diagnosis and Therapyen_HK
dc.subject.meshBase Sequenceen_HK
dc.subject.meshCarcinoma, Hepatocellular - diagnosisen_HK
dc.subject.meshHepatitis B e Antigens - analysis - geneticsen_HK
dc.subject.meshHepatitis B virus - genetics - isolation & purificationen_HK
dc.subject.meshHumansen_HK
dc.subject.meshLiver Cirrhosis - diagnosisen_HK
dc.subject.meshLiver Neoplasms - diagnosisen_HK
dc.subject.meshMolecular Sequence Dataen_HK
dc.subject.meshMutationen_HK
dc.subject.meshOligonucleotide Array Sequence Analysis - methodsen_HK
dc.subject.meshPolymorphism, Single Nucleotideen_HK
dc.subject.meshPromoter Regions, Genetic - geneticsen_HK
dc.titleSimultaneous detection of precore/basal core promoter mutations in hepatitis B virus using arrayed primer extensionen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1177-1062&volume=10&issue=2&spage=125&epage=134&date=2006&atitle=Simultaneous+detection+of+precore/basal+core+promoter+mutations+in+hepatitis+B+virus+using+arrayed+primer+extensionen_HK
dc.identifier.emailYam, WC:wcyam@hkucc.hku.hken_HK
dc.identifier.authorityYam, WC=rp00313en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.pmid16669611-
dc.identifier.scopuseid_2-s2.0-33646233432en_HK
dc.identifier.hkuros123191en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-33646233432&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume10en_HK
dc.identifier.issue2en_HK
dc.identifier.spage125en_HK
dc.identifier.epage134en_HK
dc.identifier.isiWOS:000238184700007-
dc.publisher.placeNew Zealanden_HK
dc.identifier.scopusauthoridHa, WY=7006802337en_HK
dc.identifier.scopusauthoridLau, CC=16316090100en_HK
dc.identifier.scopusauthoridYue, PYK=8570616200en_HK
dc.identifier.scopusauthoridHung, KKM=13204395900en_HK
dc.identifier.scopusauthoridChan, K=25633775800en_HK
dc.identifier.scopusauthoridChui, SH=13204613800en_HK
dc.identifier.scopusauthoridChui, AKK=7005869071en_HK
dc.identifier.scopusauthoridYam, WC=7004281720en_HK
dc.identifier.scopusauthoridWong, RNS=7402126957en_HK

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