@article{3088867,
    title = "A simplified approach for FSHD molecular testing",
    author = "Papanikos, F. and Skoulatou, C. and Sakellariou, P. and Kekou, K. and Christopoulos, T.K. and Kanavakis, E. and Traeger-Synodinos, J. and Ioannou, P.C.",
    journal = "Clinica Chimica Acta",
    year = "2014",
    volume = "429",
    pages = "96-103",
    publisher = "Elsevier B.V.",
    issn = "0009-8981",
    doi = "10.1016/j.cca.2013.11.032",
    keywords = "biological marker;  genomic DNA, article;  chromosome 10q;  chromosome 4q;  clinical article;  controlled study;  DNA base composition;  DNA determination;  facioscapulohumeral muscular dystrophy;  female;  genetic screening;  Greece;  haplotype;  human;  male;  molecular diagnostics;  nucleic acid analysis;  nucleotide sequence;  polyadenylation;  polymerase chain reaction;  population genetics;  primer extension reaction dipstick assay;  priority journal;  single nucleotide polymorphism;  telomere, Biomarkers;  FSHD;  Molecular testing, Alleles;  Base Sequence;  Chromosomes, Human, Pair 10;  Chromosomes, Human, Pair 4;  Electrophoresis;  Genome, Human;  Haplotypes;  Humans;  Molecular Diagnostic Techniques;  Muscular Dystrophy, Facioscapulohumeral;  Polymerase Chain Reaction;  Polymorphism, Genetic;  Polymorphism, Single Nucleotide;  Regulatory Sequences, Ribonucleic Acid",
    abstract = "Background: Facioscapulohumeral muscular dystrophy (FSHD) is characterized by complex genetics linked to DNA rearrangements in a polymorphic genomic region of tandemly repeated D4Z4 segments. A panel of FSHD biomarkers including contracted D4Z4 array repeat combined with the 4qA(159/161/168)PAS haplotype has been proposed as molecular signature for defining alleles causally related to FSHD. The aim of the present study was to develop a simple approach for FSHD molecular testing in order to extend studies related to the applicability of FSHD molecular signature in Greek population. Methods and results: The method comprises: (i) visual genotyping of the common 4qA and 10qA subtelomeric haplotypes by a multiplex assay in a dipstick format. (ii) Detection of 4qA161 haplotype in D4Z4 contracted alleles by tri-primer PCR. (iii) Detection of PAS SNP in PLAM region and G>C SNP in the first proximal D4Z4 unit by tri-primer PCR. The method was evaluated by analysing DNA from monoallelic sources representing common 4q and 10q haplotypes, samples from 3 FSHD families, 36 unrelated probands and 38 control individuals of Greek origin. Conclusions: The proposed method could be a very useful tool for FSHD testing making it more accessible to clinical diagnostic laboratories and the wider FSHD community. © 2013 Elsevier B.V."
}