title: Protein truncation test (PTT) to rapidly screen the DMD gene for translation terminating mutations.
abstract: We have developed a rapid and sensitive method to screen the Duchenne muscular dystrophy (DMD) mRNA for translation terminating mutations by a combination of RT-PCR (Reverse Transcription and Polymerase Chain Reaction) and in vitro transcription/translation applied to white blood cell mRNA. This technique was termed the protein truncation test (PTT). Here we demonstrate the detection of a point mutation in a DMD patient and his mother, a carrier. The PTT can also be used for carrier detection when no patient material is available, or in the case of spontaneous mutations. We developed a protocol to screen the total coding region of the DMD gene in 5-10 PTT reactions. Furthermore, PTT could be of diagnostic value in any disease where premature terminations form a substantial part of the total mutation spectrum.
authors.affiliation: Department of Human Genetics, Sylvius Laboratory, Leiden University, The Netherlands.
authors.forename: P A
authors.initials: PA
authors.lastname: Roest
authors.forename: R G
authors.initials: RG
authors.lastname: Roberts
authors.forename: A C
authors.initials: AC
authors.lastname: van der Tuijn
authors.forename: J C
authors.initials: JC
authors.lastname: Heikoop
authors.forename: G J
authors.initials: GJ
authors.lastname: van Ommen
authors.forename: J T
authors.initials: JT
authors.lastname: den Dunnen
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D016189:Dystrophin / Q000096:biosynthesis / Q000235:genetics*; D005821:Genetic Techniques; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D008969:Molecular Sequence Data; D009136:Muscular Dystrophies / Q000175:diagnosis / Q000235:genetics*; D010443:Peptide Chain Termination, Translational; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D017931:DNA Primers; D016189:Dystrophin; D012333:RNA, Messenger