Molecular analysis using DHPLC of cystic fibrosis: increase of the mutation detection rate among the affected population in Central Italy

BMC Med Genet. 2004 Apr 14:5:8. doi: 10.1186/1471-2350-5-8.

Abstract

Background: Cystic fibrosis (CF) is a multisystem disorder characterised by mutations of the CFTR gene, which encodes for an important component in the coordination of electrolyte movement across of epithelial cell membranes. Symptoms are pulmonary disease, pancreatic exocrine insufficiency, male infertility and elevated sweat concentrations. The CFTR gene has numerous mutations (>1000) and functionally important polymorphisms (>200). Early identification is important to provide appropriate therapeutic interventions, prognostic and genetic counselling and to ensure access to specialised medical services. However, molecular diagnosis by direct mutation screening has proved difficult in certain ethnic groups due to allelic heterogeneity and variable frequency of causative mutations.

Methods: We applied a gene scanning approach using DHPLC system for analysing specifically all CFTR exons and characterise sequence variations in a subgroup of CF Italian patients from the Lazio region (Central Italy) characterised by an extensive allelic heterogeneity.

Results: We have identified a total of 36 different mutations representing 88% of the CF chromosomes. Among these are two novel CFTR mutations, including one missense (H199R) and one microdeletion (4167delCTAAGCC).

Conclusion: Using this approach, we were able to increase our standard power rate of mutation detection of about 11% (77% vs. 88%).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Untranslated Regions / genetics
  • Child
  • Chromatography, High Pressure Liquid / methods*
  • Cohort Studies
  • Cystic Fibrosis / genetics*
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics
  • DNA / blood
  • DNA / genetics
  • DNA Mutational Analysis / methods*
  • DNA Mutational Analysis / trends*
  • Exons / genetics
  • Genetic Testing / methods
  • Humans
  • Introns / genetics
  • Italy
  • Leukocytes / chemistry
  • Mutation / genetics*
  • Mutation, Missense / genetics
  • Nucleic Acid Denaturation
  • Polymorphism, Genetic / genetics
  • Sequence Deletion / genetics

Substances

  • 5' Untranslated Regions
  • CFTR protein, human
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • DNA

Associated data

  • OMIM/219700
  • OMIM/602421