Lu Chao-xia, Gao Xia, Wang Jin-wei, Zhang Mei, Zhu Hong-wen, Sun Jing, Xiao Ji-fang, Yang Wei, Zhao Xiu-li, Qi Zhan, Zhang Xue
McKusick-Zhang Center for Genetic Medicine, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China.
Zhonghua Yi Xue Za Zhi. 2011 Nov 15;91(42):2971-6.
To identify the pathogenic mutations of phenylalanine hydroxylase gene (PAH) in patients with phenylketonuria (PKU) from Hebei Province.
Genomic DNA was extracted from 55 unrelated PKU patients from September 2007 to July 2009. All PAH exons and exon-intron junctions were amplified by polymerase chain reaction (PCR) and sequenced. Multiplex ligation-dependent probe amplifications (MLPA) was performed to detect the deletions or duplications of PAH. Gap-PCR was used to determine the breakpoints of large deletions.
Among them, 108 mutant alleles (98.2%) were found. All PAH exons with the exceptions of exons 9 and 13 were affected. A total of 41 different mutations were detected, including missense (n = 24), nonsense (n = 7), splicing (n = 7), small deletion (n = 1) and large deletion (n = 2). Among them, 4 missense mutations (p.Pro147Leu, p.Gly289Arg, p.Phe392Ser, p.Ile421Thr) and 2 large deletions (-4163_-406del and -1932_+3402del) were novel. The most common mutations were p.Arg243Gln (12.7%), c.611A > G (11.8%) and c.1197A > T (9.1%).
The mutations of PKU patients with from Hebei Province are scattered throughout the PAH gene. Most of them are of single nucleotide substitutions, but large deletions are not rare.
鉴定河北省苯丙酮尿症（PKU）患者苯丙氨酸羟化酶基因（PAH）的致病突变。
2007年9月至2009年7月从55例无亲缘关系的PKU患者中提取基因组DNA。通过聚合酶链反应（PCR）扩增所有PAH外显子和外显子-内含子连接区并进行测序。采用多重连接依赖探针扩增（MLPA）检测PAH的缺失或重复。采用缺口PCR确定大片段缺失的断点。
其中发现108个突变等位基因（98.2%）。除9号和13号外显子外，所有PAH外显子均受影响。共检测到41种不同突变，包括错义突变（n = 24）、无义突变（n = 7）、剪接突变（n = 7）、小缺失（n = 1）和大缺失（n = 2）。其中，4种错义突变（p.Pro147Leu、p.Gly289Arg、p.Phe392Ser、p.Ile421Thr）和2种大缺失（-4163_-406del和-1932_+3402del）为新发现的突变。最常见的突变是p.Arg243Gln（12.7%）、c.611A > G（11.8%）和c.1197A > T（9.1%）。
河北省PKU患者的突变分散在整个PAH基因中。大多数为单核苷酸替换，但大片段缺失也不少见。
