title: A simplified approach for FSHD molecular testing.
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.
  
  
  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.
authors: Laboratory of Analytical Chemistry, Department of Chemistry, Athens University, Athens 15771, Greece.
authors: Frantzeskos
authors: F
authors: Papanikos
authors: Laboratory of Analytical Chemistry, Department of Chemistry, Athens University, Athens 15771, Greece.
authors: Christina
authors: C
authors: Skoulatou
authors: Laboratory of Medical Genetics, Athens University, Athens 11527, Greece.
authors: Paraskevi
authors: P
authors: Sakellariou
authors: Laboratory of Medical Genetics, Athens University, Athens 11527, Greece.
authors: Kyriaki
authors: K
authors: Kekou
authors: Department of Chemistry, University of Patras, Patras 26500, Greece; Foundation for Research and Technology Hellas, Institute of Chemical Engineering and High Temperature Chemical Processes (FORTH/ICE-HT), Patras 26504, Greece.
authors: Theodore K
authors: TK
authors: Christopoulos
authors: Laboratory of Medical Genetics, Athens University, Athens 11527, Greece.
authors: Emmanuel
authors: E
authors: Kanavakis
authors: Laboratory of Medical Genetics, Athens University, Athens 11527, Greece.
authors: Jan
authors: J
authors: Traeger-Synodinos
authors: Laboratory of Analytical Chemistry, Department of Chemistry, Athens University, Athens 15771, Greece. Electronic address: ioannou@chem.uoa.gr.
authors: Penelope C
authors: PC
authors: Ioannou
mesh_terms: D000483:Alleles; D001483:Base Sequence; D002879:Chromosomes, Human, Pair 10 / Q000235:genetics; D002894:Chromosomes, Human, Pair 4 / Q000235:genetics; D004586:Electrophoresis; D015894:Genome, Human / Q000235:genetics; D006239:Haplotypes; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D020391:Muscular Dystrophy, Facioscapulohumeral / Q000175:diagnosis* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D020641:Polymorphism, Single Nucleotide; D038621:Regulatory Sequences, Ribonucleic Acid / Q000235:genetics
chemical_list: D038621:Regulatory Sequences, Ribonucleic Acid
keywords: Biomarkers; FSHD; Molecular testing