A wide methodological approach to identify a large duplication in CFTR gene in a CF patient uncharacterised by sequencing analysis

Lucy Costantino, Damiana Rusconi, Laura Claut, Carla Colombo, Francesca Novara, Valentina Paracchini, Luigi Porcaro, Patrizia Capasso, Orsetta Zuffardi, Manuela Seia

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Background: PCR-based diagnostic procedures are not able to characterise 6% of CF alleles. Recently, the application of array-CGH and of CFTR mRNA analysis has allowed the identification of new copy number mutations and splicing defects, that account for 2% and 13% of CF alleles, respectively, in the Italian population. Methods: Here, we report the characterisation of a large duplication in CFTR gene through different methods: MLPA assay, RT-PCR and high-resolution array-CGH. Results: We identified a large duplication, involving exons 6b-16, in a patient heterozygous for F508del mutation. This duplication produces an abnormal transcript with an out of frame addition of 2244 nucleotides and leads to the insertion of 8 amino-acid residues in the protein, followed by a stop codon. Conclusions: We propose a wide methodological approach based on MLPA assay, RT-PCR and high-resolution array-CGH to routinely analyse CF patients uncharacterised for one or both CFTR alleles.

Original languageEnglish
Pages (from-to)412-417
Number of pages6
JournalJournal of Cystic Fibrosis
Volume10
Issue number6
DOIs
Publication statusPublished - Dec 2011

Fingerprint

Alleles
Polymerase Chain Reaction
Genes
Mutation
Terminator Codon
Exons
Nucleotides
Amino Acids
Messenger RNA
Population
Proteins

Keywords

  • Array-CGH
  • CFTR
  • Duplication
  • MRNA

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Pediatrics, Perinatology, and Child Health

Cite this

A wide methodological approach to identify a large duplication in CFTR gene in a CF patient uncharacterised by sequencing analysis. / Costantino, Lucy; Rusconi, Damiana; Claut, Laura; Colombo, Carla; Novara, Francesca; Paracchini, Valentina; Porcaro, Luigi; Capasso, Patrizia; Zuffardi, Orsetta; Seia, Manuela.

In: Journal of Cystic Fibrosis, Vol. 10, No. 6, 12.2011, p. 412-417.

Research output: Contribution to journalArticle

Costantino, Lucy ; Rusconi, Damiana ; Claut, Laura ; Colombo, Carla ; Novara, Francesca ; Paracchini, Valentina ; Porcaro, Luigi ; Capasso, Patrizia ; Zuffardi, Orsetta ; Seia, Manuela. / A wide methodological approach to identify a large duplication in CFTR gene in a CF patient uncharacterised by sequencing analysis. In: Journal of Cystic Fibrosis. 2011 ; Vol. 10, No. 6. pp. 412-417.
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AU - Novara, Francesca

AU - Paracchini, Valentina

AU - Porcaro, Luigi

AU - Capasso, Patrizia

AU - Zuffardi, Orsetta

AU - Seia, Manuela

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AB - Background: PCR-based diagnostic procedures are not able to characterise 6% of CF alleles. Recently, the application of array-CGH and of CFTR mRNA analysis has allowed the identification of new copy number mutations and splicing defects, that account for 2% and 13% of CF alleles, respectively, in the Italian population. Methods: Here, we report the characterisation of a large duplication in CFTR gene through different methods: MLPA assay, RT-PCR and high-resolution array-CGH. Results: We identified a large duplication, involving exons 6b-16, in a patient heterozygous for F508del mutation. This duplication produces an abnormal transcript with an out of frame addition of 2244 nucleotides and leads to the insertion of 8 amino-acid residues in the protein, followed by a stop codon. Conclusions: We propose a wide methodological approach based on MLPA assay, RT-PCR and high-resolution array-CGH to routinely analyse CF patients uncharacterised for one or both CFTR alleles.

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