title: Molecular analysis using DHPLC of cystic fibrosis: increase of the mutation detection rate among the affected population in Central Italy.
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).
  
  
  CONCLUSIONS
  Using this approach, we were able to increase our standard power rate of mutation detection of about 11% (77% vs. 88%).
authors.affiliation: Dipartimento di Biopatologia, Università di Roma Tor Vergata, Roma, Italy. d.apice@med.uniroma2.it
authors.forename: Maria Rosaria
authors.initials: MR
authors.lastname: D'Apice
authors.forename: Stefano
authors.initials: S
authors.lastname: Gambardella
authors.forename: Mario
authors.initials: M
authors.lastname: Bengala
authors.forename: Silvia
authors.initials: S
authors.lastname: Russo
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Nardone
authors.forename: Vincenzina
authors.initials: V
authors.lastname: Lucidi
authors.forename: Federica
authors.initials: F
authors.lastname: Sangiuolo
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Novelli
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D002648:Child; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015331:Cohort Studies; D003550:Cystic Fibrosis / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004247:DNA / Q000097:blood / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods* / Q000639:trends*; D005091:Exons / Q000235:genetics; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D007438:Introns / Q000235:genetics; D007558:Italy; D007962:Leukocytes / Q000737:chemistry; D009154:Mutation / Q000235:genetics*; D020125:Mutation, Missense / Q000235:genetics; D009691:Nucleic Acid Denaturation; D011110:Polymorphism, Genetic / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics
chemical_list: D020121:5' Untranslated Regions; C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA