Spinocerebellar Ataxia Tethering PCR: A Rapid Genetic Test for the Diagnosis of Spinocerebellar Ataxia Types 1, 2, 3, 6, and 7 by PCR and Capillary Electrophoresis.
The Journal of Molecular Diagnostics. 2018-05-01; 20(3): 289-297
DOI: 10.1016/j.jmoldx.2017.12.006
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1. J Mol Diagn. 2018 May;20(3):289-297. doi: 10.1016/j.jmoldx.2017.12.006. Epub 2018
Feb 17.
Spinocerebellar Ataxia Tethering PCR: A Rapid Genetic Test for the Diagnosis of
Spinocerebellar Ataxia Types 1, 2, 3, 6, and 7 by PCR and Capillary
Electrophoresis.
Cagnoli C(1), Brussino A(1), Mancini C(1), Ferrone M(2), Orsi L(3), Salmin P(4),
Pappi P(4), Giorgio E(1), Pozzi E(1), Cavalieri S(1), Di Gregorio E(1), Ferrero
M(1), Filla A(5), De Michele G(5), Gellera C(6), Mariotti C(6), Nethisinghe S(7),
Giunti P(7), Stevanin G(8), Brusco A(9).
Author information:
(1)Department of Medical Sciences, University of Turin, Turin, Italy.
(2)Department of Medical Sciences, University of Turin, Turin, Italy; Medical
Genetics Unit, Città della Salute e della Scienza University Hospital, Turin,
Italy.
(3)Department of Laboratory Medicine, and the Neurologic Division I, Department
of Neuroscience and Mental Health, Città della Salute e della Scienza University
Hospital, Turin, Italy.
(4)Medical Genetics Unit, Città della Salute e della Scienza University Hospital,
Turin, Italy.
(5)Department of Neurosciences, Odontostomatological and Reproductive Sciences,
University Federico II, Naples, Italy.
(6)Unit of Genetics of Neurodegenerative and Metabolic Diseases, Fondazione IRCCS
Carlo Besta Neurological Institute, Milan, Italy.
(7)Ataxia Centre, Department of Molecular Neuroscience, Institute of Neurology,
University College London, London, United Kingdom.
(8)INSERM, U 1127, Institut du Cerveau et de la Moelle epinière, Paris, France;
Centre National de la Recherche Scientifique UMR 7225, Paris, France; UMRS 1127,
Université Pierre et Marie Curie (Paris 06), Sorbonne Universités, Paris, France;
Ecole Pratique des Hautes Etudes, PSL Research University, Paris, France; Centre
de Référence de Neurogénétique, Hôpital de la Pitié-Salpêtrière, Assistance
Publique – Hôpitaux de Paris, Paris, France.
(9)Department of Medical Sciences, University of Turin, Turin, Italy; Medical
Genetics Unit, Città della Salute e della Scienza University Hospital, Turin,
Italy. Electronic address: .
Spinocerebellar ataxia (SCA) types 1, 2, 3, 6, and 7, associated with a (CAG)n
repeat expansion in coding sequences, are the most prevalent autosomal dominant
ataxias worldwide (approximately 60% of the cases). In addition, the phenotype of
SCA2 expansions has been now extended to Parkinson disease and amyotrophic
lateral sclerosis. Their diagnosis is currently based on a PCR to identify small
expanded alleles, followed by a second-level test whenever a false normal
homozygous or a CAT interruption in SCA1 needs to be verified. Next-generation
sequencing still does not allow efficient detection of these repeats. Here, we
show the efficacy of a novel, rapid, and cost-effective method to identify and
size pathogenic expansions in SCA1, 2, 3, 6, and 7 and recognize large alleles or
interruptions without a second-level test. Twenty-five healthy controls and 33
expansion carriers were analyzed: alleles migrated consistently in different PCRs
and capillary runs, and homozygous individuals were always distinguishable from
heterozygous carriers of both common and large (>100 repeats) pathogenic CAG
expansions. Repeat number could be calculated counting the number of peaks,
except for the largest SCA2 and SCA7 alleles. Interruptions in SCA1 were always
visible. Overall, our method allows a simpler, cost-effective, and sensibly
faster SCA diagnostic protocol compared with the standard technique and to the
still unadapted next-generation sequencing.
Copyright © 2018 American Society for Investigative Pathology and the Association
for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
DOI: 10.1016/j.jmoldx.2017.12.006
PMID: 29462666 [Indexed for MEDLINE]