NF1 gene analysis based on DHPLC.

Alessandro De Luc, Anna Buccino, Debora Gianni, Massimo Mangino, Sandra Giustini, Antonio Richetta, Luigina Divona, Stefano Calvieri, Rita Mingarelli, Bruno Dallapiccola

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The high mutation rate at the NF1 locus results in a wide range of molecular abnormalities. The majority of these mutations are private and rare, generating elevated allelic diversity with a restricted number of recurrent mutations. In this study, we have assessed the efficacy of denaturing high-performance liquid chromatography (DHPLC), for detecting mutation in the NF1 gene. DHPLC is a fast and highly sensitive technique based on the detection of heteroduplexes in PCR products by ion pair reverse-phase HPLC under partially denaturing conditions. We established theoretical conditions for DHPLC analysis of all coding exons and splice junctions of the NF1 gene using the WAVEmaker software version 4.1.40 and screened for mutations a panel of 40 unrelated NF1 patients (25 sporadic and 15 familial), genetically uncharacterized. Disruptive mutations were identified in 29 individuals with an overall mutation detection rate of 72.5%. The mutations included eight deletions (exons 4b, 7, 10a, 14, 26, and 31), one insertion (exon 8), nine nonsense mutation (exons 10a, 13, 23.1, 27a, 29, 31, and 36), six missense mutations (exons 15, 16, 17, 24, and 31), four splice errors (exons 11, 14, 36, and 40) and a complex rearrangement within exon 16. Eighteen (62%) of the identified disruptive mutations are novel. Seven unclassified and three previously reported polymorphisms were also detected. None of the missense mutations identified in this study were found after screening of 150 controls. Our results suggest that DHPLC provides an accurate method for the rapid identification of NF1 mutations.

Original languageEnglish
Pages (from-to)171-172
Number of pages2
JournalHuman Mutation
Volume21
Issue number2
Publication statusPublished - Feb 2003

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Exons
High Pressure Liquid Chromatography
Mutation
Genes
Mutation Rate
Missense Mutation
Nonsense Codon
Software
Ions
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

De Luc, A., Buccino, A., Gianni, D., Mangino, M., Giustini, S., Richetta, A., ... Dallapiccola, B. (2003). NF1 gene analysis based on DHPLC. Human Mutation, 21(2), 171-172.

NF1 gene analysis based on DHPLC. / De Luc, Alessandro; Buccino, Anna; Gianni, Debora; Mangino, Massimo; Giustini, Sandra; Richetta, Antonio; Divona, Luigina; Calvieri, Stefano; Mingarelli, Rita; Dallapiccola, Bruno.

In: Human Mutation, Vol. 21, No. 2, 02.2003, p. 171-172.

Research output: Contribution to journalArticle

De Luc, A, Buccino, A, Gianni, D, Mangino, M, Giustini, S, Richetta, A, Divona, L, Calvieri, S, Mingarelli, R & Dallapiccola, B 2003, 'NF1 gene analysis based on DHPLC.', Human Mutation, vol. 21, no. 2, pp. 171-172.
De Luc A, Buccino A, Gianni D, Mangino M, Giustini S, Richetta A et al. NF1 gene analysis based on DHPLC. Human Mutation. 2003 Feb;21(2):171-172.
De Luc, Alessandro ; Buccino, Anna ; Gianni, Debora ; Mangino, Massimo ; Giustini, Sandra ; Richetta, Antonio ; Divona, Luigina ; Calvieri, Stefano ; Mingarelli, Rita ; Dallapiccola, Bruno. / NF1 gene analysis based on DHPLC. In: Human Mutation. 2003 ; Vol. 21, No. 2. pp. 171-172.
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