河北地区55例苯丙酮尿症患者苯丙氨酸羟化酶基因突变的检测与分析  

Mutation analysis of phenylalanine hydroxylase gene in 55 patients with phenylketonuria from Hebei Province

作　　者：卢超霞[1] 高峡 王金玮[3] 张梅 朱宏文[1] 孙婧[1] 肖继芳[1] 杨威[1] 赵秀丽[1] 齐展[1] 张学[1] 

机构地区：[1]中国医学科学院基础医学研究所北京协和医学院基础医学院医学遗传学系,北京100005 [2]河北省张家口市妇幼保健院 [3]河北省廊坊市妇幼保健中心

出　　处：《中华医学杂志》2011年第42期2971-2976,共6页National Medical Journal of China

基　　金：国家高技术研究发展“863”项目（2007AA022440）

摘　　要：目的了解河北地区苯丙酮尿症（PKU）患者苯丙氨酸羟化酶（PAH）基因突变情况，分析该地区PAH基因的突变谱及突变特点。方法联合应用PAH基因全部外显子及外显子一内含子交界区扩增测序和多重连接依赖式探针扩增（MLPA）方法，对2007年9月至2009年7月就诊的55例来自河北地区的PKU患者进行PAH基因点突变和拷贝数改变的检测。对发现大片段缺失的个体，用跨越断裂位点（Gap）-PCR-测序方法确定断裂点精确位置。结果110个PAH等位基因中检出突变等位基因108个，突变检出率为98．2％。共发现PAH基因突变41种，涉及11个外显子，包括错义突变24种、无义突变7种、剪接位点突变7种、小的缺失1种和大片段缺失2种，其中4种错义突变（P．Pr0147Leu，P．Gly289Arg，P．Phe392Ser，P．Ile421Thr）和2种大片段缺失突变（-4163_-406del和-1932一十3402del）为国际首次报道。55例患者中以下3种突变所占比例最高：P．Arg243Gln（12．7％）、e．611A〉G（11．8％）和c．1197A〉T（9．1％）。结论中国河北地区PKU患者中PAH基因的突变分布广泛；突变类型大部分为单碱基替换，但是大片段缺失也占一定比例。Objective To identify the pathogenic mutations of phenylalanine hydroxylase gene （PAH） in patients with phenylketonuria （PKU） from Hebei Province. Methods 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. Results 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% ）. Conclusion 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.

关 键 词：苯丙酮尿症 苯丙氨酸羟化酶 突变谱 多重连接依赖式探针扩增 

分 类 号：R725.8[医药卫生—儿科]