NF2 mutation screening by denaturing high-performance liquid chromatography and high-resolution melting analysis
| Autor: | Maurizio Genuardi, Costanza Bacci, Claudio Orlando, Aldesia Provenzano, Laura Papi, Roberta Sestini |
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| Rok vydání: | 2008 |
| Předmět: |
Neurofibromatosis 2
TRANSMISSION DNA Mutational Analysis MOSAICISM Biology Settore MED/03 - GENETICA MEDICA Nucleic Acid Denaturation Polymerase Chain Reaction Sensitivity and Specificity High Resolution Melt Denaturing high performance liquid chromatography CONDUCTANCE REGULATOR GENE NEUROFIBROMATOSIS 2 DHPLC CANCER TYPE-2 IDENTIFICATION POLYMORPHISMS PREVALENCE Exon Germline mutation Genes Neurofibromatosis 2 otorhinolaryngologic diseases Humans Point Mutation Transition Temperature Multiplex NF2 mutation screening high-resolution melting analysis Gene Genetics (clinical) Chromatography High Pressure Liquid DNA Primers Genetics Point mutation Exons Neuroma Acoustic |
| Zdroj: | Genetic testing. 12(2) |
| ISSN: | 1090-6576 |
| Popis: | Neurofibromatosis type 2 (NF2) is an autosomal-dominant disorder caused by mutations in the NF2 gene and predisposing to the development of nervous system. Identification of germline mutations is essential to provide appropriate genetic counseling in NF2 patients, but it represents an extremely challenging task because the vast majority of mutations are unique and spread over the entire coding sequence. Moreover, about 30% of de novo patients are indeed mosaic, and direct sequencing can undetect mutated alleles present in a minority of cells. As most screening techniques do not meet the requirements for efficient NF2 testing, we have developed a semi-automated denaturing high-performance liquid chromatography (DHPLC) method for point mutation detection combined with a multiplex ligation-dependent probe amplification approach to screen for gene rearrangements. In addition, we have evaluated high-resolution melting analysis (HRMA) as an exon scanning procedure to identify point mutations in the NF2 gene. The results obtained in 92 NF2 patients expand the NF2 mutational spectrum and indicate DHPLC and HRMA as good systems to screen for point mutations in diseases with a heterogeneous spectrum of alterations. |
| Databáze: | OpenAIRE |
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