Protein truncation test (PTT) to rapidly screen the DMD gene for translation terminating mutations

Neuromuscul Disord. 1993 Sep-Nov;3(5-6):391-4. doi: 10.1016/0960-8966(93)90083-v.

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.

Publication types

  • Case Reports
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA Primers
  • Dystrophin / biosynthesis
  • Dystrophin / genetics*
  • Genetic Techniques
  • Humans
  • Infant, Newborn
  • Male
  • Molecular Sequence Data
  • Muscular Dystrophies / diagnosis
  • Muscular Dystrophies / genetics*
  • Peptide Chain Termination, Translational*
  • Point Mutation*
  • Polymerase Chain Reaction / methods
  • Protein Biosynthesis
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism*
  • Transcription, Genetic

Substances

  • DNA Primers
  • Dystrophin
  • RNA, Messenger