The Need for Establishing a Universal CTG Sizing Method in Myotonic Dystrophy Type 1

Genes

Alejandro Lucia¹  SarahA. Cumming²  DarrenG. Monckton²  NadiaM. Murillo-Melo³  JonathanJ. Magaña³  Alicia Martínez-Piñeiro⁴  Gisela Nogales-Gadea⁴  Emma Koehorst⁴  Judit Núñez-Manchón⁴  Guillem Pintos-Morell⁴  Ian Linares-Pardo⁴  Giuseppe Lucente⁴  Alfonsina Ballester-Lopez⁴  Alba Ramos-Fransi⁴  Miriam Almendrote⁴  Andrea Arbex⁴  Jaume Coll-Cantí⁴ 

[1] Faculty of Sport Sciences, Madrid, Spain, & Instituto de Investigación Hospital 12 de Octubre (imas12), Universidad Europea, 28041 Madrid, Spain;Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G128QQ, UK;Laboratory of Genomic Medicine, Department of Genetics, National Rehabilitation Institute (INR-LGII), 14389 Mexico City, Mexico;Neuromuscular and Neuropediatric Research Group, Institut d’Investigació en Ciències de la Salut Germans Trias i Pujol, Campus Can Ruti, Universitat Autònoma de Barcelona, 08916 Badalona, Barcelona, Spain;

关键词: CTG expansion size;    myotonic dystrophy type 1;    long PCR;    small pool-PCR;    age of disease onset;   

DOI  :  10.3390/genes11070757

来源: DOAJ

【 摘 要 】

The number of cytosine-thymine-guanine (CTG) repeats (‘CTG expansion size’) in the 3′untranslated region (UTR) region of the dystrophia myotonica-protein kinase (DMPK) gene is a hallmark of myotonic dystrophy type 1 (DM1), which has been related to age of disease onset and clinical severity. However, accurate determination of CTG expansion size is challenging due to its characteristic instability. We compared five different approaches (heat pulse extension polymerase chain reaction [PCR], long PCR-Southern blot [with three different primers sets—1, 2 and 3] and small pool [SP]-PCR) to estimate CTG expansion size in the progenitor allele as well as the most abundant CTG expansion size, in 15 patients with DM1. Our results indicated variability between the methods (although we found no overall differences between long PCR 1 and 2 and SP-PCR, respectively). While keeping in mind the limited sample size of our patient cohort, SP-PCR appeared as the most suitable technique, with an inverse significant correlation found between CTG expansion size of the progenitor allele, as determined by this method, and age of disease onset (r = −0.734, p = 0.016). Yet, in light of the variability of the results obtained with the different methods, we propose that an international agreement is needed to determine which is the most suitable method for assessing CTG expansion size in DM1.

【 授权许可】

Unknown