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Research Article
Rapid and reliable detection of exon rearrangements in various movement disorders genes by multiplex ligation-dependent probe amplification

Published on-line Movement Disorders. 2 Aug 07

Ana Djarmati, PhD 1 2, Miodrag Guvi, MSc 1 2 3, Anne Grünewald, MSc 1 2, Anthony E. Lang, MD 4, Peter P. Pramstaller, MD 5, David K. Simon, MD, PhD 6, Angela M. Kaindl, MD 7 8, Peter Vieregge, MD 1 9, Anders O. H. Nygren, PhD 10, Christian Beetz, PhD 11, Katja Hedrich, PhD 1 2, Christine Klein, MD 1

* Keywords multiplex ligation-dependent probe amplification • Parkinson's disease • dopa-responsive dystonia • myoclonus-dystonia • exon rearrangements


Abstract

Because of the occurrence of different types of mutations, comprehensive genetic testing for Parkinson's disease (PD), dopa-responsive dystonia (DRD), and myoclonus-dystonia (M-D) should include screening for small sequence changes and for large exonic rearrangements in disease-associated genes. In diagnostic and research settings, the latter is frequently omitted or performed by laborious and expensive quantitative real-time PCR (qPCR). Our study aimed to evaluate the utility of a novel method, multiplex ligation-dependent probe amplification (MLPA), in molecular diagnostics of movement disorders. We have analyzed, by MLPA, genomic DNA from 21 patients affected with PD, DRD, or M-D, in which the presence of exon rearrangement(s) (n = 20) or of a specific point mutation (detectable by MLPA, n = 1) had been established previously by qPCR or sequencing. In parallel, we have studied, in a blinded fashion, DNA from 49 patients with an unknown mutational status. Exon rearrangements were evident in 20 samples with previously established mutations; in the 21st sample the known specific point mutation was detected. We conclude that MLPA represents a reliable method for large-scale and cost-effective gene dosage screening of various movement disorders genes. This finding reaches far beyond a simple technical advancement and has two major implications: (1) By improving the availability of comprehensive genetic testing, it supports clinicians in the establishment of a genetically defined diagnosis; (2) By enabling gene dosage testing of several genes simultaneously, it significantly facilitates the mutational analysis of large patient and control populations and thereby constitutes the prerequisite for meaningful phenotype-genotype correlations.

Published Online: 2 Aug 2007

© 2007 Movement Disorder Society