Use of multiplex PCR and CE for gene dosage quantification and its biomedical application for SMN, PMP22, and α-globin genes
Journal
Electrophoresis
Journal Volume
28
Journal Issue
16
Pages
2826-2834
Date Issued
2007
Author(s)
Hung C.-C.
Chien S.-C.
Lin C.-Y.
Chang C.-H.
Chang Y.-F.
Jong Y.-J.
Hsieh W.-S.
Liu M.S.
Su Y.-N.
Abstract
Many genetic diseases are caused by the presence of point mutations, small insertions, and deletions in respective genes, and the number of diseases known to be caused by deletions and duplications involving large DNA genomes is increasing. These changes lead to underexpression or overexpression of the gene, according to changes in gene dosage. The methods for the detection of point mutations, small insertions, and deletions are well established, but the detection of larger genomic deletions or duplications is more difficult. Due to the lack of efficient and technically feasible protocols for gene dosage quantification, we describe a diagnostic protocol employing a combination of available methods. The efficient and accurate gene dosage quantification platform is combined with multiplex PCR and CE, and applied to detect dosages of several genes, including SMN, PMP22, and α-globin genes. The reliability of this novel methodology shows that it is a relatively speedy and low-cost procedure and a significant tool for genetic diagnosis. Its sensitivity and specificity for identifying deletion and duplication genotypes approach 100%. Moreover, once we establish this powerful system, we will further apply this technique to the rapid detection of trisomy syndromes and microdeletion syndromes, including trisomy 13, Down syndrome, DiGeorge syndrome, and others. ? 2007 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
SDGs
Other Subjects
alpha globin; peripheral myelin protein 22; survival motor neuron protein; article; capillary electrophoresis; clinical article; controlled study; diagnostic accuracy; DiGeorge syndrome; Down syndrome; gene deletion; gene dosage; gene duplication; gene identification; gene insertion; genetic code; genetic screening; genome analysis; genotype; human; multiplex polymerase chain reaction; mutational analysis; point mutation; reliability; trisomy; trisomy 13; Base Sequence; Cyclic AMP Response Element-Binding Protein; DNA; DNA Primers; Electrophoresis, Capillary; Gene Dosage; Genomics; Globins; Humans; Myelin Proteins; Nerve Tissue Proteins; Polymerase Chain Reaction; RNA-Binding Proteins
Type
journal article