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Use of multiplex PCR and CE for gene dosage quantification and its biomedical applications for SMN , PMP22 , and α ‐globin genes
Author(s) -
Hung ChiaCheng,
Chien ShuChin,
Lin ChiaYun,
Chang ChienHui,
Chang YinFei,
Jong YuhJyh,
Hsieh SungTsang,
Hsieh WuShiun,
Liu Ming S.,
Lin WinLi,
Lee ChienNan,
Su YiNing
Publication year - 2007
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200600647
Subject(s) - gene dosage , gene duplication , biology , genetics , copy number variation , gene , trisomy , multiplex , multiplex polymerase chain reaction , computational biology , genome , point mutation , multiplex ligation dependent probe amplification , polymerase chain reaction , mutation , gene expression , exon
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.