File Download

There are no files associated with this item.

  Links for fulltext
     (May Require Subscription)
Supplementary

Article: Allele dropout in PCR-based diagnosis of Wilson disease: Mechanisms and solutions

TitleAllele dropout in PCR-based diagnosis of Wilson disease: Mechanisms and solutions
Authors
Issue Date2006
PublisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.org
Citation
Clinical Chemistry, 2006, v. 52 n. 3, p. 517-520 How to Cite?
AbstractBackground: We investigated the mechanisms leading to allele dropout-the nonamplification of 1 of the alleles-in PCR-based diagnosis of Wilson disease (WD). Methods: We extracted genomic DNA from blood samples from 6 WD patients (P1-P6) with allele dropouts detected in a previous study of WD in a Hong Kong Chinese population. We amplified the ATP7B gene by PCR and performed direct DNA sequencing of all exons of the ATP7B gene. To support the proposed mechanism of allele dropout, we used proof leading DNA polymerase, primer design avoiding single-nucleotide polymorphism sites, and duplex PCR. Results: Patients P1-P4 were all apparently homozygous for a known disease-causing mutation, c.2975C>T (p.P992L) in exon 13. Patient P5 was apparently homozygous for a novel mutation, c.2524G>A, and patient P6 was apparently homozygous for another known mutation, c.522_523insA (p.K175K-fs). In all cases, we determined that the patients were actually heterozygous for these mutations. Conclusion: Our results confirm that allele dropout is the mechanism causing apparent homozygosity of heterozygous mutations in these WD patients. © 2006 American Association for Clinical Chemistry.
Persistent Identifierhttp://hdl.handle.net/10722/148450
ISSN
2015 Impact Factor: 7.457
2015 SCImago Journal Rankings: 2.472
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLam, CWen_US
dc.contributor.authorMak, CMen_US
dc.date.accessioned2012-05-29T06:13:03Z-
dc.date.available2012-05-29T06:13:03Z-
dc.date.issued2006en_US
dc.identifier.citationClinical Chemistry, 2006, v. 52 n. 3, p. 517-520en_US
dc.identifier.issn0009-9147en_US
dc.identifier.urihttp://hdl.handle.net/10722/148450-
dc.description.abstractBackground: We investigated the mechanisms leading to allele dropout-the nonamplification of 1 of the alleles-in PCR-based diagnosis of Wilson disease (WD). Methods: We extracted genomic DNA from blood samples from 6 WD patients (P1-P6) with allele dropouts detected in a previous study of WD in a Hong Kong Chinese population. We amplified the ATP7B gene by PCR and performed direct DNA sequencing of all exons of the ATP7B gene. To support the proposed mechanism of allele dropout, we used proof leading DNA polymerase, primer design avoiding single-nucleotide polymorphism sites, and duplex PCR. Results: Patients P1-P4 were all apparently homozygous for a known disease-causing mutation, c.2975C>T (p.P992L) in exon 13. Patient P5 was apparently homozygous for a novel mutation, c.2524G>A, and patient P6 was apparently homozygous for another known mutation, c.522_523insA (p.K175K-fs). In all cases, we determined that the patients were actually heterozygous for these mutations. Conclusion: Our results confirm that allele dropout is the mechanism causing apparent homozygosity of heterozygous mutations in these WD patients. © 2006 American Association for Clinical Chemistry.en_US
dc.languageengen_US
dc.publisherAmerican Association for Clinical Chemistry, Inc. The Journal's web site is located at http://www.clinchem.orgen_US
dc.relation.ispartofClinical Chemistryen_US
dc.subject.meshAdenosine Triphosphatases - Geneticsen_US
dc.subject.meshAllelesen_US
dc.subject.meshCation Transport Proteins - Geneticsen_US
dc.subject.meshHepatolenticular Degeneration - Diagnosis - Geneticsen_US
dc.subject.meshHeterozygoteen_US
dc.subject.meshHomozygoteen_US
dc.subject.meshHumansen_US
dc.subject.meshMutationen_US
dc.subject.meshPolymerase Chain Reactionen_US
dc.subject.meshPolymorphism, Single Nucleotideen_US
dc.titleAllele dropout in PCR-based diagnosis of Wilson disease: Mechanisms and solutionsen_US
dc.typeArticleen_US
dc.identifier.emailLam, CW:ching-wanlam@pathology.hku.hken_US
dc.identifier.authorityLam, CW=rp00260en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1373/clinchem.2005.060491en_US
dc.identifier.pmid16510432en_US
dc.identifier.scopuseid_2-s2.0-33644514003en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-33644514003&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume52en_US
dc.identifier.issue3en_US
dc.identifier.spage517en_US
dc.identifier.epage520en_US
dc.identifier.isiWOS:000235674900025-
dc.publisher.placeUnited Statesen_US

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats