Molecular analysis of PKU-associated PAH mutations: a fast and simple genotyping test
Autor: | Amelia Pasquali, Antonio Angeloni, Vincenzo Leuzzi, Sirio D'Amici, Cristiana Artiola, Carla Carducci, Teresa Giovanniello, Claudia Carducci, Manuela Tolve, Antonio Pizzuti |
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Jazyk: | angličtina |
Rok vydání: | 2018 |
Předmět: |
Genetics
phenylalanine hydroxylase deficiency Phenylalanine hydroxylase biology PKU screening New variant Biochemistry Genetics and Molecular Biology (miscellaneous) Italian population Molecular analysis Structural Biology PKU mutation analysis Genotype biology.protein Technical Note Phenylketonuria Carrier screening Dried blood Genotyping Biotechnology |
Zdroj: | Methods and Protocols |
Popis: | Neonatal screening for phenylketonuria (PKU, OMIM: 261600) was introduced at the end of the 1960s. We developed a rapid and simple molecular test for the most frequent phenylalanine hydroxylase (PAH, Gene ID: 5053) mutations. Using this method to detect the 18 most frequent mutations, it is possible to achieve a 75% detection rate in Italian population. The variants selected also reach a high detection rate in other populations, for example, 70% in southern Germany, 68% in western Germany, 76% in Denmark, 68% in Sweden, 63% in Poland, and 60% in Bulgaria. We successfully applied this confirmation test in neonatal screening for hyperphenylalaninemias using dried blood spots and obtained the genotype in approximately 48 h. The method was found to be suitable as second tier test in neonatal screening for hyperphenylalaninemias in neonates with a positive screening test. This test can also be useful for carrier screening because it can bypass the entire coding sequence and intron–exon boundaries sequencing, thereby overcoming the questions that this approach implies, such as new variant interpretations. |
Databáze: | OpenAIRE |
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