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Simple, efficient, and cost-effective multiplex genotyping with matrix assisted laser desorption/ionization time-of-flight mass spectrometry of hemoglobin beta gene mutations

dc.contributor.authorWanna Thongnoppakhunen_US
dc.contributor.authorSurasak Jiemsupen_US
dc.contributor.authorSuganya Yongkiettrakulen_US
dc.contributor.authorChompunut Kanjanakornen_US
dc.contributor.authorChanin Limwongseen_US
dc.contributor.authorPrapon Wilairaten_US
dc.contributor.authorAnusorn Vanasanten_US
dc.contributor.authorNanyawan Rungrojen_US
dc.contributor.authorPa Thai Yenchitsomanusen_US
dc.contributor.otherMahidol Universityen_US
dc.contributor.otherThailand National Center for Genetic Engineering and Biotechnologyen_US
dc.date.accessioned2018-09-13T06:28:14Z
dc.date.available2018-09-13T06:28:14Z
dc.date.issued2009-01-01en_US
dc.description.abstractA number of common mutations in the hemoglobin β (HBB) gene cause β-thalassemia, a monogenic disease with high prevalence in certain ethnic groups. As there are 30 HBB variants that cover more than 99.5% of HBB mutant alleles in the Thai population, an efficient and cost-effective screening method is required. Three panels of multiplex primer extensions, followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were developed. The first panel simultaneously detected 21 of the most common HBB mutations, while the second panel screened nine additional mutations, plus seven of the first panel for confirmation; the third panel was used to confirm three HBB mutations, yielding a 9-Da mass difference that could not be clearly distinguished by the previous two panels. The protocol was both standardized using 40 samples of known genotypes and subsequently validated in 162 blind samples with 27 different genotypes (including a normal control), comprising heterozygous, compound heterozygous, and homozygous β-thalassemia. Results were in complete agreement with those from the genotyping results , conducted using three different methods overall. The method developed here permitted the detection of mutations missed using a single genotyping procedure. The procedure should serve as the method of choice for HBB genotyping due to its accuracy, sensitivity, and cost-effectiveness, and can be applied to studies of other gene variants that are potential disease biomarkers. Copyright © American Society for Investigative Pathology and the Association for Molecular Pathology.en_US
dc.identifier.citationJournal of Molecular Diagnostics. Vol.11, No.4 (2009), 334-346en_US
dc.identifier.doi10.2353/jmoldx.2009.080151en_US
dc.identifier.issn15251578en_US
dc.identifier.other2-s2.0-69249138783en_US
dc.identifier.urihttps://repository.li.mahidol.ac.th/handle/123456789/27326
dc.rightsMahidol Universityen_US
dc.rights.holderSCOPUSen_US
dc.source.urihttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=69249138783&origin=inwarden_US
dc.subjectBiochemistry, Genetics and Molecular Biologyen_US
dc.subjectMedicineen_US
dc.titleSimple, efficient, and cost-effective multiplex genotyping with matrix assisted laser desorption/ionization time-of-flight mass spectrometry of hemoglobin beta gene mutationsen_US
dc.typeArticleen_US
dspace.entity.typePublication
mu.datasource.scopushttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=69249138783&origin=inwarden_US
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