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.
Calì, F., Ruggeri, G., Vinci, M., Meli C, Carducci C, Leuzzi V, et al. (2010). Exon deletions of the PAH gene in Italian hyperphenylalaninemics. EXPERIMENTAL AND MOLECULAR MEDICINE, 42(2) [10.3858/emm.2010.42.2.009].
Exon deletions of the PAH gene in Italian hyperphenylalaninemics
VINCI, Mirella;MICCICHE', Salvatore;ROMANO, Valentino
2010-01-01
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.File | Dimensione | Formato | |
---|---|---|---|
emm-42-81.pdf
Solo gestori archvio
Dimensione
553.25 kB
Formato
Adobe PDF
|
553.25 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.