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

Publisher: BioMed Central Country of Publication: England NLM ID: 100968552 Publication Model: Electronic Cited Medium: Internet ISSN: 1471-2350 (Electronic) Linking ISSN: 14712350 NLM ISO Abbreviation: BMC Med Genet Subsets: MEDLINE

Original Publication: London : BioMed Central, [2000-

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

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%).

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OMIM 219700; 602421

0 (5' Untranslated Regions)
0 (CFTR protein, human)
126880-72-6 (Cystic Fibrosis Transmembrane Conductance Regulator)
9007-49-2 (DNA)

Date Created: 20040416 Date Completed: 20040527 Latest Revision: 20220317

20240104

PMC419352

10.1186/1471-2350-5-8

15084222