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Article: Rapid diagnosis of Wilson disease by a 28-mutation panel: Real-time amplification refractory mutation system in diagnosing acute Wilsonian liver failure

TitleRapid diagnosis of Wilson disease by a 28-mutation panel: Real-time amplification refractory mutation system in diagnosing acute Wilsonian liver failure
Authors
Issue Date2008
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/cca
Citation
Clinica Chimica Acta, 2008, v. 398 n. 1-2, p. 39-42 How to Cite?
AbstractBackground: Wilson disease is one of the commonest inherited and potentially fatal yet treatable liver disorders. About 5-27% patients present with acute liver failure and require prompt chelation therapy and life-saving liver transplantation. Diagnosis during acute liver failure is particularly difficult with short time allowance. Direct molecular diagnosis remains the most decisive tool but is often hindered by demanding techniques and numerous mutations. We developed a one-step, 3-h, reproducible, and accurate real-time amplification refractory mutation system which can simultaneously detect 28 ATP7B mutations. Methods: Primers were designed to complement the mutant sequence at the 3′ end. The mutations were p.S105X, p.Q511X, p.R616Q, p.S693P, p.S693C, p.R778L, p.A874V, p.T888P, p.R919G, p.T935M, p.P992L, p.M1025R, p.D1047V, p.I1148T, p.R1156H, p.T1178A, p.V1216M, p.P1273Q, p.G1281C, p.R1320S, p.V1334D, p.V176SfsX28, p.G869EfsX4, IVS3+1G>T, IVS4-1G>C, IVS4-5T>G, IVS6+9A>G, and IVS9+5G>T. Reaction was performed using QuantiTect SYBR Green PCR Master Mix on an Applied Biosystems StepOne thermal cycler. Values of the threshold cycle were compared between normal and mutant alleles. Results: Primers of all mutations were highly specific with absence of wild-type amplification. All the results were validated by direct DNA sequencing. Conclusions: This rapid and cost-efficient method allows wide mutation coverage, rendering the SYBR-green assay feasible and attractive for large-scale routine application. © 2008 Elsevier B.V. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/148586
ISSN
2015 Impact Factor: 2.799
2015 SCImago Journal Rankings: 1.040
ISI Accession Number ID
Funding AgencyGrant Number
Research Grants Council of the Hong Kong Special Administrative Region, ChinaCUHK4084/02M
Chan Woon Cheung Education and Research Fund
Funding Information:

The work described in this paper was partially supported by a grant from the Research Grants Council of the Hong Kong Special Administrative Region, China (Project no. CUHK4084/02M) and Chan Woon Cheung Education and Research Fund.

References

 

DC FieldValueLanguage
dc.contributor.authorMak, CMen_US
dc.contributor.authorLam, CWen_US
dc.contributor.authorLai, STen_US
dc.contributor.authorHui, Yen_US
dc.contributor.authorTam, Sen_US
dc.date.accessioned2012-05-29T06:13:55Z-
dc.date.available2012-05-29T06:13:55Z-
dc.date.issued2008en_US
dc.identifier.citationClinica Chimica Acta, 2008, v. 398 n. 1-2, p. 39-42en_US
dc.identifier.issn0009-8981en_US
dc.identifier.urihttp://hdl.handle.net/10722/148586-
dc.description.abstractBackground: Wilson disease is one of the commonest inherited and potentially fatal yet treatable liver disorders. About 5-27% patients present with acute liver failure and require prompt chelation therapy and life-saving liver transplantation. Diagnosis during acute liver failure is particularly difficult with short time allowance. Direct molecular diagnosis remains the most decisive tool but is often hindered by demanding techniques and numerous mutations. We developed a one-step, 3-h, reproducible, and accurate real-time amplification refractory mutation system which can simultaneously detect 28 ATP7B mutations. Methods: Primers were designed to complement the mutant sequence at the 3′ end. The mutations were p.S105X, p.Q511X, p.R616Q, p.S693P, p.S693C, p.R778L, p.A874V, p.T888P, p.R919G, p.T935M, p.P992L, p.M1025R, p.D1047V, p.I1148T, p.R1156H, p.T1178A, p.V1216M, p.P1273Q, p.G1281C, p.R1320S, p.V1334D, p.V176SfsX28, p.G869EfsX4, IVS3+1G>T, IVS4-1G>C, IVS4-5T>G, IVS6+9A>G, and IVS9+5G>T. Reaction was performed using QuantiTect SYBR Green PCR Master Mix on an Applied Biosystems StepOne thermal cycler. Values of the threshold cycle were compared between normal and mutant alleles. Results: Primers of all mutations were highly specific with absence of wild-type amplification. All the results were validated by direct DNA sequencing. Conclusions: This rapid and cost-efficient method allows wide mutation coverage, rendering the SYBR-green assay feasible and attractive for large-scale routine application. © 2008 Elsevier B.V. All rights reserved.en_US
dc.languageengen_US
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/ccaen_US
dc.relation.ispartofClinica Chimica Actaen_US
dc.subject.meshAdulten_US
dc.subject.meshAllelesen_US
dc.subject.meshCost-Benefit Analysisen_US
dc.subject.meshDna - Geneticsen_US
dc.subject.meshDna Mutational Analysis - Economics - Methodsen_US
dc.subject.meshDna Primersen_US
dc.subject.meshFemaleen_US
dc.subject.meshFluorescent Dyesen_US
dc.subject.meshHepatolenticular Degeneration - Complications - Diagnosis - Geneticsen_US
dc.subject.meshHumansen_US
dc.subject.meshLiver Failure, Acute - Diagnosis - Etiology - Geneticsen_US
dc.subject.meshMutation - Geneticsen_US
dc.subject.meshOrganic Chemicalsen_US
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen_US
dc.titleRapid diagnosis of Wilson disease by a 28-mutation panel: Real-time amplification refractory mutation system in diagnosing acute Wilsonian liver failureen_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.doi10.1016/j.cca.2008.08.002en_US
dc.identifier.pmid18760268-
dc.identifier.scopuseid_2-s2.0-54049104289en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-54049104289&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume398en_US
dc.identifier.issue1-2en_US
dc.identifier.spage39en_US
dc.identifier.epage42en_US
dc.identifier.isiWOS:000261074200008-
dc.publisher.placeNetherlandsen_US

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