A rapid and reliable detection system for the analysis of PMP22 gene dosage by MP/DHPLC assay
Journal
Journal of Human Genetics
Journal Volume
51
Journal Issue
3
Pages
227-235
Date Issued
2006
Author(s)
Abstract
Charcot-Marie-Tooth disease type 1A (CMT1A) and hereditary neuropathy with liability to pressure palsies (HNPP) are caused by a 1.5-Mb duplication and a deletion at chromosome 17p11.2-12 encompassing the peripheral myelin protein 22 gene (PMP22), respectively. We developed a rapid and reliable detection system for duplications/deletions of the PMP22 gene based on measurement of gene copy number. The method involves amplification of a test locus with unknown copy number and a reference locus of known copy number by multiplex PCR (MP), followed by denaturing high-performance liquid chromatography (DHPLC) or capillary electrophoresis detection to identify single copy changes. Thirty-two patients with CMT1A, 17 patients with HNPP, and 61 unaffected individuals were analyzed. Using the same competitive MP protocol, the measured PMP22 gene dosage revealed concordant results between DHPLC and capillary electrophoresis analysis. The results of the MP/DHPLC or the MP/capillary electrophoresis assay were all confirmed by PCR-restriction fragment length polymorphism analysis. We concluded that the MP/ DHPLC assay is an efficient, accurate, and reliable technique for gene dosage determination of the PMP22 gene for CMT1A duplication and HNPP deletion. This technique further extends the application of DHPLC as an alternative method for the measurement of gene amplifications and heterozygous deletions in different genetic diseases. ? The Japan Society of Human Genetics and Springer-Verlag 2006.
SDGs
Other Subjects
gene product; peripheral myelin protein 22; analytic method; article; capillary electrophoresis; clinical article; comparative study; controlled study; CYBB gene; denaturing high performance liquid chromatography; diagnostic accuracy; diagnostic value; gene; gene amplification; gene deletion; gene dosage; gene duplication; genetic analysis; hereditary motor sensory neuropathy; hereditary neuropathy with liability to pressure palsies; heterozygosity; human; KRIT1 gene; multiplex polymerase chain reaction; neuropathy; nucleotide sequence; PMP22 gene; reliability; restriction fragment length polymorphism; Base Sequence; Chromatography, High Pressure Liquid; DNA Primers; Electrophoresis, Capillary; Gene Dosage; Humans; Myelin Proteins; Reproducibility of Results
Type
journal article