Mutation detection remains problematic for large genes, primarily because PCR-based methodology fails to detect heterozygous deletions and any duplication. In the ATM gene only a handful of multi-exon deletions have been described to date, and this type of mutation has been considered rare. To address this issue we tested a new MLPA (Multiplex Ligation Probe Amplification) kit that covers 33 of the 66 ATM exons, using for controls two previously characterized genomic deletions in addition to three A-T patients, taken from a survey of nine, who had missing four mutations unidentified after conventional mutation screening. We identified for the first time: 1) a approximately 41 kb genomic duplication spanning exons 4-20 (c.-30_2816dup41kb)(a.k.a., ATM dup 41 kb); 2) a novel genomic deletion including exon 31, and 3) in hemizygosis a point mutation in the non-deleted exon 31. In this study we extended mutation detection to nine new Italian A-T patients, using a combined approach of haplotype analysis, DHPLC and MLPA. Overall we achieved a mutation detection rate of >97%, and can now define a spectrum of ATM mutations based on twenty-one consecutive Italian families with A-T.

Large Genomic Mutations within the ATM Gene Detected by MLPA, Including a Duplication of 41 kb from Exon 4 to 20

CAVALIERI, Simona;FUNARO, Ada;MIGONE, Nicola;BRUSCO, Alfredo
2008-01-01

Abstract

Mutation detection remains problematic for large genes, primarily because PCR-based methodology fails to detect heterozygous deletions and any duplication. In the ATM gene only a handful of multi-exon deletions have been described to date, and this type of mutation has been considered rare. To address this issue we tested a new MLPA (Multiplex Ligation Probe Amplification) kit that covers 33 of the 66 ATM exons, using for controls two previously characterized genomic deletions in addition to three A-T patients, taken from a survey of nine, who had missing four mutations unidentified after conventional mutation screening. We identified for the first time: 1) a approximately 41 kb genomic duplication spanning exons 4-20 (c.-30_2816dup41kb)(a.k.a., ATM dup 41 kb); 2) a novel genomic deletion including exon 31, and 3) in hemizygosis a point mutation in the non-deleted exon 31. In this study we extended mutation detection to nine new Italian A-T patients, using a combined approach of haplotype analysis, DHPLC and MLPA. Overall we achieved a mutation detection rate of >97%, and can now define a spectrum of ATM mutations based on twenty-one consecutive Italian families with A-T.
2008
72
10
18
http://onlinelibrary.wiley.com/doi/10.1111/j.1469-1809.2007.00399.x/abstract;jsessionid=3FA9B4E0FD18CF2687D239DF31086EF6.f03t03
ATM; Ataxia-telangiectasia; Multiple Ligation Probe Amplification; large genomic mutations; ATM duplication; Italian mutation spectrum
CAVALIERI S; FUNARO A; PAPPI P; MIGONE N; GATTI RA; BRUSCO A
File in questo prodotto:
File Dimensione Formato  
49 Large Genomic Mutations within the ATM Gene Detected by MLPA_Ann Hum Genet 2008.pdf

Accesso riservato

Descrizione: Paper pubblicato
Tipo di file: PDF EDITORIALE
Dimensione 784.17 kB
Formato Adobe PDF
784.17 kB Adobe PDF   Visualizza/Apri   Richiedi una copia

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2318/34299
Citazioni
  • ???jsp.display-item.citation.pmc??? 8
  • Scopus 28
  • ???jsp.display-item.citation.isi??? 26
social impact