Leiden Muscular Dystrophy pages^©

Molecular diagnosis for DMD / BMD

(last modified March 19, 2006)

Contents

• Diagnosis of other neuromusuclar diseases
• Molecular diagnostic tests for neuromuscular disorders in Leiden 
  
• Molecular diagnosis DMD/BMD
  • general information
  • protocols from Leiden
• DNA analysis
  • my opinion, techniques available
• RNA analysis
  • DMD PTT, cDNA sequencing, tissue-specific transcription, etc.
• Protein analysis
• Cell line analysis - MyoD-induced myo-differentiation
• Biopsy analysis

Diagnosis: general information

• diagnosis/epidemiology of neuromuscular disorders: European Neuro-Muscular Centre (ENMC)
  • diagnostic criteria - DMD/BMD (chapter 1)
• EMQN  -  The European Molecular Genetics Quality Network
• EuroGenTest  -  European network for genetic test development
• EDDNAL  -  European directory of laboratories performing DNA diagnosis
• CMGS  -  Clinical Molecular Genetics Society (UK)
  to advance the science of clinical molecular genetics and to further public education
• Concerted Action on Genetic Services in Europe (CAGSE): Genetic Enquiry Center Manchester

Diagnostic protocols from Leiden

In this section, links are provided to the certified protocols we use in Leiden in our DNA diagnostic department, headed by dr. Bert Bakker.

• General
  • buffers, solutions
  • DNA isolation
    • from blood
    • from chorion villi
    • phenol extraction
  • Southern blotting
  • hybridization (incl. probe labeling)
  • Protein Truncation Test (PTT)
• DMD specific
  • introduction / flow-chart
  • multiplex PCR: detection deletions
  • detection deletions and duplications
    • Southern blotting / cDNA hybridization
    • quantitative multiplex PCR
  • haplotype analysis in/around the DMD-gene
  • RT-PCR and Protein Truncation Test: detection point mutations

DNA-analysis

RNA analysis

Protein analysis

• Western blotting
  • multiplex Western blot  (coming soon)
• immunohistochemistry
• myoD-induced myodifferentiation
• antibodies
  • dystrophin antibodies and their epitopes
  • dystroglycan and sarcoglycan antibodies
  • antibodies to dystrophin related proteins

Cell line analysis

• MyoD-induced myo-differentiation - the technology
  • myoD-gene delivery - adenoviral / retroviral
  • examples / figures