Τίτλος:
A simplified approach for FSHD molecular testing
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
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.
Συγγραφείς:
Papanikos, F.
Skoulatou, C.
Sakellariou, P.
Kekou, K.
Christopoulos, T.K.
Kanavakis, E.
Traeger-Synodinos, J.
Ioannou, P.C.
Περιοδικό:
Clinica Chimica Acta
Λέξεις-κλειδιά:
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
DOI:
10.1016/j.cca.2013.11.032
