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 language | English |
|---|---|
| Pages (from-to) | 412-417 |
| Number of pages | 6 |
| Journal | Journal of Cystic Fibrosis |
| Volume | 10 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Dec 2011 |
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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 journal › Article
}
TY - JOUR
T1 - A wide methodological approach to identify a large duplication in CFTR gene in a CF patient uncharacterised by sequencing analysis
AU - Costantino, Lucy
AU - Rusconi, Damiana
AU - Claut, Laura
AU - Colombo, Carla
AU - Novara, Francesca
AU - Paracchini, Valentina
AU - Porcaro, Luigi
AU - Capasso, Patrizia
AU - Zuffardi, Orsetta
AU - Seia, Manuela
PY - 2011/12
Y1 - 2011/12
N2 - 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.
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.
KW - Array-CGH
KW - CFTR
KW - Duplication
KW - MRNA
UR - http://www.scopus.com/inward/record.url?scp=81455141390&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=81455141390&partnerID=8YFLogxK
U2 - 10.1016/j.jcf.2011.06.007
DO - 10.1016/j.jcf.2011.06.007
M3 - Article
C2 - 21852204
AN - SCOPUS:81455141390
VL - 10
SP - 412
EP - 417
JO - Journal of Cystic Fibrosis
JF - Journal of Cystic Fibrosis
SN - 1569-1993
IS - 6
ER -