title: A multiplexed ARMS-PCR approach for the detection of common MECP2 mutations.
abstract: Rett syndrome (RTT), an X-linked dominant neurodevelopmental disorder, is caused mainly by de novo mutations in the methyl-CpG-binding protein 2 gene (MECP2). Although more than 200 different MECP2 mutations have been identified throughout the gene, 7 of those (p.R133C, p.T158M, p.R168X, p.R255X, p.R270X, p.R294X, and p.R306C) account for up to two-thirds of pathogenic mutations in RTT patients. A rapid and efficient screening strategy for these mutations can be used as a preliminary step for genetic diagnosis of RTT. The current protocols used for this purpose are of high cost and require special equipment. We have designed a simpler multiplex amplification refractory mutation system (ARMS)-PCR strategy that allows identification of these common MECP2 mutant alleles in four PCR reactions. The assay was tested in 14 RTT patients who were previously genotyped using PCR-restriction fragment length polymorphism and DNA sequencing. A complete concordance was observed between the results of the two methods. The multiplex ARMS-PCR does not require any special equipment, and it provides rapid, reproducible, and cost-effective detection of common MECP2 mutations. The assay can be carried out efficiently in a standard molecular genetics laboratory and suitable as a preliminary screen for all patients with RTT diagnosis.
authors.affiliation: Department of Molecular Biology and Genetics, Bogazici University, Istanbul, Turkey. ibrahim.baris@boun.edu.tr
authors.forename: Ibrahim
authors.initials: I
authors.lastname: Baris
authors.forename: Esra
authors.initials: E
authors.lastname: Battaloglu
mesh_terms: D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D051783:Methyl-CpG-Binding Protein 2 / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015518:Rett Syndrome / Q000175:diagnosis / Q000235:genetics*
chemical_list: D017931:DNA Primers; C497392:MECP2 protein, human; D051783:Methyl-CpG-Binding Protein 2