Multiplex ligation-dependent probe amplification assay for simultaneous detection of Parkinson's disease gene rearrangements

Authors

  • Oronzo Scarciolla MD,

    1. Aging Research Center, CeSi, University G. d'Annunzio, Chieti, Italy
    2. Department of Biomedical Sciences, University G. d'Annunzio, Chieti, Italy
    3. Department of Clinical Sciences and Bioimaging, University G. d'Annunzio, Chieti-Pescara, Italy
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  • Francesco Brancati MD,

    1. Aging Research Center, CeSi, University G. d'Annunzio, Chieti, Italy
    2. Department of Biomedical Sciences, University G. d'Annunzio, Chieti, Italy
    3. IRCCS CSS-Mendel Institute, Rome, Italy
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  • Enza Maria Valente MD, PhD,

    1. IRCCS CSS-Mendel Institute, Rome, Italy
    2. Operative Unit of Pediatric Genetics and Immunology, Department of Medical and Surgical Pediatric Sciences, University of Messina, Messina, Italy
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  • Alessandro Ferraris MD,

    1. IRCCS CSS-Mendel Institute, Rome, Italy
    2. Department of Experimental Medicine and Pathology, University La Sapienza, Rome, Italy
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  • Maria Vittoria De Angelis MD,

    1. Neurodegenerative Diseases Unit, Aging Research Center, CeSi, University G. d'Annunzio, Chieti, Italy
    2. Department of Human Motor Sciences, University G. d'Annunzio, Chieti, Italy
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  • Stefano Valbonesi BPh,

    1. Aging Research Center, CeSi, University G. d'Annunzio, Chieti, Italy
    2. Department of Biomedical Sciences, University G. d'Annunzio, Chieti, Italy
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  • Barbara Garavaglia PhD,

    1. Molecular Neurogenetics Unit, Foundation IRCCS Neurological Institute “C. Besta”, Milan, Italy
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  • Antonino Uncini MD,

    1. Neurodegenerative Diseases Unit, Aging Research Center, CeSi, University G. d'Annunzio, Chieti, Italy
    2. Department of Human Motor Sciences, University G. d'Annunzio, Chieti, Italy
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  • Giandomenico Palka MD,

    1. Aging Research Center, CeSi, University G. d'Annunzio, Chieti, Italy
    2. Department of Biomedical Sciences, University G. d'Annunzio, Chieti, Italy
    3. Human Genetics Division, Hospital of Pescara, Pescara, Italy
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  • Liborio Stuppia MD,

    Corresponding author
    1. Aging Research Center, CeSi, University G. d'Annunzio, Chieti, Italy
    2. Department of Biomedical Sciences, University G. d'Annunzio, Chieti, Italy
    3. IGM CNR, Bologna, Italy
    • Department of Biomedical Sciences, University Gabriele d'Annunzio, Via dei Vestini, 31, 66013 Chieti, Italy
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  • Bruno Dallapiccola MD

    1. IRCCS CSS-Mendel Institute, Rome, Italy
    2. Operative Unit of Pediatric Genetics and Immunology, Department of Medical and Surgical Pediatric Sciences, University of Messina, Messina, Italy
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Abstract

Parkinson's disease (PD) is a common disorder caused by degeneration of dopaminergic neurons in the substantia nigra and other brain areas. Mutations in several genes have been associated with both autosomal dominant PD and recessive early onset Parkinsonism (EOP). Genomic rearrangements such as deletions or multiplications of one or more exons represent a common mutational mechanism for most of these genes and are not detectable with routine mutation screening techniques. MLPA (Multiplex Ligation-dependent Probe Amplification), is a cheap, simple, rapid, and sensitive tool to detect exon dosage alterations and specific point mutations in selected genes. We tested the recently developed PD-MLPA assay by using 13 positive control samples carrying known mutations in SNCA, LRRK2, Parkin, PINK1, and DJ-1 genes. We then applied this technique to screen 16 EOP patients who were then cross-tested by quantitative PCR (qPCR). All the mutations present in the positive control samples were clearly detected by MLPA. Moreover, three novel Parkin rearrangements were identified among EOP patients and confirmed by qPCR. Only two samples generated false positive duplications of LRRK2 exon 1 and UCH-L1 exon 9, respectively. These results show that PD-MLPA assay can simultaneously and effectively detect rearrangements in most PD genes (SNCA, Parkin, PINK1, and DJ-1) as well as the LRRK2 G2019S common mutation. Thus, the use of this novel platform can improve the analysis of such mutations, facilitating comprehensive genetic testing in PD and EOP. © 2007 Movement Disorder Society

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