Tolve Manuela, Artiola Cristiana, Pasquali Amelia, Giovanniello Teresa, D'Amici Sirio, Angeloni Antonio, Pizzuti Antonio, Carducci Claudia, Leuzzi Vincenzo, Carducci Carla
Department of Experimental Medicine, Sapienza University, 00185 Rome, Italy.
Department of Human Neuroscience, Child Neurology and Psychiatry Sapienza University, 00185 Rome, Italy.
Methods Protoc. 2018 Aug 16;1(3):30. doi: 10.3390/mps1030030.
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 (, 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.
新生儿苯丙酮尿症（PKU，OMIM：261600）筛查始于20世纪60年代末。我们针对最常见的苯丙氨酸羟化酶（基因ID：5053）突变开发了一种快速简便的分子检测方法。使用该方法检测18种最常见的突变，在意大利人群中检测率可达75%。所选变体在其他人群中也有较高的检测率，例如在德国南部为70%，在德国西部为68%，在丹麦为76%，在瑞典为68%，在波兰为63%，在保加利亚为60%。我们成功地将这种确证检测应用于使用干血斑进行的高苯丙氨酸血症新生儿筛查，并在大约48小时内获得基因型。该方法被认为适合作为新生儿高苯丙氨酸血症筛查阳性新生儿的二线检测方法。该检测对于携带者筛查也很有用，因为它可以绕过整个编码序列和内含子-外显子边界测序，从而克服这种方法所带来的问题，如新变体解释等。
