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Methods Protoc. 2018, 1(3), 30; https://doi.org/10.3390/mps1030030

Molecular Analysis of PKU-Associated PAH Mutations: A Fast and Simple Genotyping Test

1
Department of Experimental Medicine, Sapienza University, 00185 Rome, Italy
2
Department of Human Neuroscience, Child Neurology and Psychiatry Sapienza University, 00185 Rome, Italy
*
Author to whom correspondence should be addressed.
Received: 20 June 2018 / Revised: 6 August 2018 / Accepted: 6 August 2018 / Published: 16 August 2018
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Abstract

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. View Full-Text
Keywords: phenylketonuria; phenylalanine hydroxylase deficiency; PKU mutation analysis; PKU screening phenylketonuria; phenylalanine hydroxylase deficiency; PKU mutation analysis; PKU screening
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Tolve, M.; Artiola, C.; Pasquali, A.; Giovanniello, T.; D’Amici, S.; Angeloni, A.; Pizzuti, A.; Carducci, C.; Leuzzi, V.; Carducci, C. Molecular Analysis of PKU-Associated PAH Mutations: A Fast and Simple Genotyping Test. Methods Protoc. 2018, 1, 30.

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