Exon deletions of the PAH gene in Italian hyperphenylalaninemics

Valentino Romano, Mirella Vinci, Salvatore Micciche', Alda Ragalmuto, Valeria Chiavetta, Vincenzo Leuzzi, Concetta Meli, Pietro Schinocca, Carla Carducci, Simone Pozzessere, Valentino Romano, Giuseppa Ruggeri, Mirella Vinci, Francesco Calì

Risultato della ricerca: Article

16 Citazioni (Scopus)

Abstract

A consistent finding of many studies describing the spectrum of mutant phenylalanine hydroxylase alleles underlying hyperphenylalaninemia is the impossibility of achieving a 100% mutation ascertainment rate using conventional gene-scanning methods such as Denaturing Gradient Gel Electrophoresis (DGGE), Denaturing High Performance Liquid Chromatography (DHPLC), direct sequencing. In recent years, it has been shown that a significant proportion of these undetermined alleles consist of large deletions overlapping one or more exons. These deletions have been difficult to detect in compound heterozygotes by gene-scanning methods due to the masking effect by the non-deleted allele. To date no systematic search has been carried out for such exon deletions in Italian patients with phenylketonuria or Mild hyperphenylalaninemia. We have used Multiplex Ligation-dependent Probe Amplification (MLPA), Comparative Multiplex Dosage Analysis (CMDA) and Real- Time PCR to search for both large deletions and duplications of the phenylalanine hydroxylase gene in Italian HPA patients. Overall, 4 deletions removing different Phenylalanine Hydroxylase (PAH) gene exons were identified in 12 patients. Two of these deletions involving Exons 4-5-6-7-8 (systematic name c.353-?_912+?del) and Exon 6 (systematic name c.510-?_706+?del) have not been reported previously. In this study we show that exon deletion of the PAH gene account for 1.7% of all mutant PAH alleles in Italian hyperphenylalaninemics. A consistent finding of many studies describing the spectrum of mutant phenylalanine hydroxylase alleles underlying hyperphenylalaninemia is the impossibility of achieving a 100% mutation ascertainment rate using conventional gene-scanning methods such as Denaturing Gradient Gel Electrophoresis (DGGE), Denaturing High Performance Liquid Chromatography (DHPLC), direct sequencing. In recent years, it has been shown that a significant proportion of these undetermined alleles consist of large deletions overlapping one or more exons. These deletions have been difficult to detect in compound heterozygotes by gene-scanning methods due to the masking effect by the non-deleted allele. To date no systematic search has been carried out for such exon deletions in Italian patients with phenylketonuria or Mild hyperphenylalaninemia. We have used Multiplex Ligation-dependent Probe Amplification (MLPA), Comparative Multiplex Dosage Analysis (CMDA) and Real- Time PCR to search for both large deletions and duplications of the phenylalanine hydroxylase gene in Italian HPA patients. Overall, 4 deletions removing different Phenylalanine Hydroxylase (PAH) gene exons were identified in 12 patients. Two of these deletions involving Exons 4-5-6-7-8 (systematic name c.353-?_912+?del) and Exon 6 (systematic name c.510-?_706+?del) have not been reported previously. In this study we show that exon deletion of the PAH gene account for 1.7% of all mutant PAH alleles in Italian hyperphenylalaninemics.
Lingua originaleEnglish
pagine (da-a)-
Numero di pagine0
RivistaExperimental and Molecular Medicine
Volume42
Stato di pubblicazionePublished - 2010

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Phenylalanine Hydroxylase
Exons
Genes
Phenylketonurias
Alleles
Names
Scanning
Denaturing Gradient Gel Electrophoresis
Multiplex Polymerase Chain Reaction
High performance liquid chromatography
Mutation Rate
Heterozygote
Electrophoresis
Amplification
Real-Time Polymerase Chain Reaction
Gels
High Pressure Liquid Chromatography

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Clinical Biochemistry
  • Molecular Biology
  • Molecular Medicine

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Exon deletions of the PAH gene in Italian hyperphenylalaninemics. / Romano, Valentino; Vinci, Mirella; Micciche', Salvatore; Ragalmuto, Alda; Chiavetta, Valeria; Leuzzi, Vincenzo; Meli, Concetta; Schinocca, Pietro; Carducci, Carla; Pozzessere, Simone; Romano, Valentino; Ruggeri, Giuseppa; Vinci, Mirella; Calì, Francesco.

In: Experimental and Molecular Medicine, Vol. 42, 2010, pag. -.

Risultato della ricerca: Article

Romano, V, Vinci, M, Micciche', S, Ragalmuto, A, Chiavetta, V, Leuzzi, V, Meli, C, Schinocca, P, Carducci, C, Pozzessere, S, Romano, V, Ruggeri, G, Vinci, M & Calì, F 2010, 'Exon deletions of the PAH gene in Italian hyperphenylalaninemics', Experimental and Molecular Medicine, vol. 42, pagg. -.
Romano, Valentino ; Vinci, Mirella ; Micciche', Salvatore ; Ragalmuto, Alda ; Chiavetta, Valeria ; Leuzzi, Vincenzo ; Meli, Concetta ; Schinocca, Pietro ; Carducci, Carla ; Pozzessere, Simone ; Romano, Valentino ; Ruggeri, Giuseppa ; Vinci, Mirella ; Calì, Francesco. / Exon deletions of the PAH gene in Italian hyperphenylalaninemics. In: Experimental and Molecular Medicine. 2010 ; Vol. 42. pagg. -.
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abstract = "A consistent finding of many studies describing the spectrum of mutant phenylalanine hydroxylase alleles underlying hyperphenylalaninemia is the impossibility of achieving a 100{\%} mutation ascertainment rate using conventional gene-scanning methods such as Denaturing Gradient Gel Electrophoresis (DGGE), Denaturing High Performance Liquid Chromatography (DHPLC), direct sequencing. In recent years, it has been shown that a significant proportion of these undetermined alleles consist of large deletions overlapping one or more exons. These deletions have been difficult to detect in compound heterozygotes by gene-scanning methods due to the masking effect by the non-deleted allele. To date no systematic search has been carried out for such exon deletions in Italian patients with phenylketonuria or Mild hyperphenylalaninemia. We have used Multiplex Ligation-dependent Probe Amplification (MLPA), Comparative Multiplex Dosage Analysis (CMDA) and Real- Time PCR to search for both large deletions and duplications of the phenylalanine hydroxylase gene in Italian HPA patients. Overall, 4 deletions removing different Phenylalanine Hydroxylase (PAH) gene exons were identified in 12 patients. Two of these deletions involving Exons 4-5-6-7-8 (systematic name c.353-?_912+?del) and Exon 6 (systematic name c.510-?_706+?del) have not been reported previously. In this study we show that exon deletion of the PAH gene account for 1.7{\%} of all mutant PAH alleles in Italian hyperphenylalaninemics. A consistent finding of many studies describing the spectrum of mutant phenylalanine hydroxylase alleles underlying hyperphenylalaninemia is the impossibility of achieving a 100{\%} mutation ascertainment rate using conventional gene-scanning methods such as Denaturing Gradient Gel Electrophoresis (DGGE), Denaturing High Performance Liquid Chromatography (DHPLC), direct sequencing. In recent years, it has been shown that a significant proportion of these undetermined alleles consist of large deletions overlapping one or more exons. These deletions have been difficult to detect in compound heterozygotes by gene-scanning methods due to the masking effect by the non-deleted allele. To date no systematic search has been carried out for such exon deletions in Italian patients with phenylketonuria or Mild hyperphenylalaninemia. We have used Multiplex Ligation-dependent Probe Amplification (MLPA), Comparative Multiplex Dosage Analysis (CMDA) and Real- Time PCR to search for both large deletions and duplications of the phenylalanine hydroxylase gene in Italian HPA patients. Overall, 4 deletions removing different Phenylalanine Hydroxylase (PAH) gene exons were identified in 12 patients. Two of these deletions involving Exons 4-5-6-7-8 (systematic name c.353-?_912+?del) and Exon 6 (systematic name c.510-?_706+?del) have not been reported previously. In this study we show that exon deletion of the PAH gene account for 1.7{\%} of all mutant PAH alleles in Italian hyperphenylalaninemics.",
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T1 - Exon deletions of the PAH gene in Italian hyperphenylalaninemics

AU - Romano, Valentino

AU - Vinci, Mirella

AU - Micciche', Salvatore

AU - Ragalmuto, Alda

AU - Chiavetta, Valeria

AU - Leuzzi, Vincenzo

AU - Meli, Concetta

AU - Schinocca, Pietro

AU - Carducci, Carla

AU - Pozzessere, Simone

AU - Romano, Valentino

AU - Ruggeri, Giuseppa

AU - Vinci, Mirella

AU - Calì, Francesco

PY - 2010

Y1 - 2010

N2 - A consistent finding of many studies describing the spectrum of mutant phenylalanine hydroxylase alleles underlying hyperphenylalaninemia is the impossibility of achieving a 100% mutation ascertainment rate using conventional gene-scanning methods such as Denaturing Gradient Gel Electrophoresis (DGGE), Denaturing High Performance Liquid Chromatography (DHPLC), direct sequencing. In recent years, it has been shown that a significant proportion of these undetermined alleles consist of large deletions overlapping one or more exons. These deletions have been difficult to detect in compound heterozygotes by gene-scanning methods due to the masking effect by the non-deleted allele. To date no systematic search has been carried out for such exon deletions in Italian patients with phenylketonuria or Mild hyperphenylalaninemia. We have used Multiplex Ligation-dependent Probe Amplification (MLPA), Comparative Multiplex Dosage Analysis (CMDA) and Real- Time PCR to search for both large deletions and duplications of the phenylalanine hydroxylase gene in Italian HPA patients. Overall, 4 deletions removing different Phenylalanine Hydroxylase (PAH) gene exons were identified in 12 patients. Two of these deletions involving Exons 4-5-6-7-8 (systematic name c.353-?_912+?del) and Exon 6 (systematic name c.510-?_706+?del) have not been reported previously. In this study we show that exon deletion of the PAH gene account for 1.7% of all mutant PAH alleles in Italian hyperphenylalaninemics. A consistent finding of many studies describing the spectrum of mutant phenylalanine hydroxylase alleles underlying hyperphenylalaninemia is the impossibility of achieving a 100% mutation ascertainment rate using conventional gene-scanning methods such as Denaturing Gradient Gel Electrophoresis (DGGE), Denaturing High Performance Liquid Chromatography (DHPLC), direct sequencing. In recent years, it has been shown that a significant proportion of these undetermined alleles consist of large deletions overlapping one or more exons. These deletions have been difficult to detect in compound heterozygotes by gene-scanning methods due to the masking effect by the non-deleted allele. To date no systematic search has been carried out for such exon deletions in Italian patients with phenylketonuria or Mild hyperphenylalaninemia. We have used Multiplex Ligation-dependent Probe Amplification (MLPA), Comparative Multiplex Dosage Analysis (CMDA) and Real- Time PCR to search for both large deletions and duplications of the phenylalanine hydroxylase gene in Italian HPA patients. Overall, 4 deletions removing different Phenylalanine Hydroxylase (PAH) gene exons were identified in 12 patients. Two of these deletions involving Exons 4-5-6-7-8 (systematic name c.353-?_912+?del) and Exon 6 (systematic name c.510-?_706+?del) have not been reported previously. In this study we show that exon deletion of the PAH gene account for 1.7% of all mutant PAH alleles in Italian hyperphenylalaninemics.

AB - A consistent finding of many studies describing the spectrum of mutant phenylalanine hydroxylase alleles underlying hyperphenylalaninemia is the impossibility of achieving a 100% mutation ascertainment rate using conventional gene-scanning methods such as Denaturing Gradient Gel Electrophoresis (DGGE), Denaturing High Performance Liquid Chromatography (DHPLC), direct sequencing. In recent years, it has been shown that a significant proportion of these undetermined alleles consist of large deletions overlapping one or more exons. These deletions have been difficult to detect in compound heterozygotes by gene-scanning methods due to the masking effect by the non-deleted allele. To date no systematic search has been carried out for such exon deletions in Italian patients with phenylketonuria or Mild hyperphenylalaninemia. We have used Multiplex Ligation-dependent Probe Amplification (MLPA), Comparative Multiplex Dosage Analysis (CMDA) and Real- Time PCR to search for both large deletions and duplications of the phenylalanine hydroxylase gene in Italian HPA patients. Overall, 4 deletions removing different Phenylalanine Hydroxylase (PAH) gene exons were identified in 12 patients. Two of these deletions involving Exons 4-5-6-7-8 (systematic name c.353-?_912+?del) and Exon 6 (systematic name c.510-?_706+?del) have not been reported previously. In this study we show that exon deletion of the PAH gene account for 1.7% of all mutant PAH alleles in Italian hyperphenylalaninemics. A consistent finding of many studies describing the spectrum of mutant phenylalanine hydroxylase alleles underlying hyperphenylalaninemia is the impossibility of achieving a 100% mutation ascertainment rate using conventional gene-scanning methods such as Denaturing Gradient Gel Electrophoresis (DGGE), Denaturing High Performance Liquid Chromatography (DHPLC), direct sequencing. In recent years, it has been shown that a significant proportion of these undetermined alleles consist of large deletions overlapping one or more exons. These deletions have been difficult to detect in compound heterozygotes by gene-scanning methods due to the masking effect by the non-deleted allele. To date no systematic search has been carried out for such exon deletions in Italian patients with phenylketonuria or Mild hyperphenylalaninemia. We have used Multiplex Ligation-dependent Probe Amplification (MLPA), Comparative Multiplex Dosage Analysis (CMDA) and Real- Time PCR to search for both large deletions and duplications of the phenylalanine hydroxylase gene in Italian HPA patients. Overall, 4 deletions removing different Phenylalanine Hydroxylase (PAH) gene exons were identified in 12 patients. Two of these deletions involving Exons 4-5-6-7-8 (systematic name c.353-?_912+?del) and Exon 6 (systematic name c.510-?_706+?del) have not been reported previously. In this study we show that exon deletion of the PAH gene account for 1.7% of all mutant PAH alleles in Italian hyperphenylalaninemics.

KW - Comparative Multiplex Dosage Analysis

UR - http://hdl.handle.net/10447/44599

M3 - Article

VL - 42

SP - -

JO - Experimental and Molecular Medicine

JF - Experimental and Molecular Medicine

SN - 1226-3613

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