Denaturing HPLC-based approach for detection of COL7A1 gene mutations causing dystrophic epidermolysis bullosa

Patrizia Posteraro, Monica Pascucci, Marina Colombi, Sergio Barlati, Alberto Giannetti, Mauro Paradisi, Aki Mustonen, Giovanna Zambruno, Daniele Castiglia

Research output: Contribution to journalArticlepeer-review


Dystrophic epidermolysis bullosa (DEB) is a rare clinically heterogeneous genodermatosis due to genetic defects in type VII collagen gene (COL7A1). Identification of COL7A1 mutations is a challenge since this gene comprises 118 exons and more than 300 mutations scattered over the gene have been reported. Here, we describe for the first time the use of denaturing high performance liquid chromatography (DHPLC) for COL7A1 mutation detection. To validate the method, exon-specific DHPLC conditions were applied to screen DNA samples from patients carrying known COL7A1 mutations. Abnormal DHPLC profiles were obtained for all known mutations. Subsequent DHPLC analysis of 17 DEB families of unknown genotype allowed the identification of 21 distinct mutations, 9 of which were novel. The DHPLC mutation detection rate was significantly higher compared with our mutation scanning rate with conventional techniques (97% vs 86%), indicating DHPLC as the method of choice for COL7A1 molecular characterization in DEB patients.

Original languageEnglish
Pages (from-to)1391-1401
Number of pages11
JournalBiochemical and Biophysical Research Communications
Issue number3
Publication statusPublished - Dec 23 2005


  • COL7A1
  • Denaturing high performance liquid chromatography
  • Dystrophic epidermolysis bullosa
  • Mutation detection
  • Type VII collagen

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology


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