摘要
目的:检测苯丙酮尿症(PKU)患者苯丙氨酸羟化酶(PAH)基因突变谱,探讨其突变率与临床严重度的相关性。方法:53例PKU患者根据其治疗前的血苯丙氨酸浓度进行临床分型,同时提取其基因组DNA,PCR扩增PAH基因外显子3、5、6、7、10、11和12,用单链构像多态分析、变性高效液相层析(DHPLC)和直接测序3种方法筛查PAH基因突变。用Kendall等级相关分析临床严重度和突变基因型的相关性。结果:4种突变(R176X、E280K、L367R和S349A)在中国PKU人群中未见报道,其中2种突变(L367R和S349A)为国际首次报道。基因型和临床严重度之间具有较好的相关性(r=0.696,P<0.01)。结论:新发现的PKU患者的4种基因新突变,丰富了PKU患者PAH基因突变谱。基因型和临床严重度间较好的相关性为临床上PKU的基因诊断提供了重要的理论依据。DHPLC法是一种快速、准确、经济的筛选PAH基因突变的新方法。
Objective：To determine the spectrum of 53 Chinese with phenylketonuria （PKU）phenylalanine hydroxylase（PAH）mutations and explore the con＇elation between the genotype and the clinical severity.Methods： The pretreatment blood phe concentration of 53 PKU patients was detected by high-performance liquid chro- matography.The clinical severity was classified to three types,the severe type （phe concentration〉1200μmol/ L）,the moderate type （600-1200μmol/ L） and the mild type （〈600μmol/ L）.Genomic DNA was isolated from perpheral blood samples of 53 patients.PAH gene mutation screening analyse were performed by polymerase chain reactions （PCR）of exon 3,5,6,7,10,11 and 12,followed by single strand conformation polymorphism （SSCP）or denaturing high-performance liquid chromatography （DHPLC）.The genotypes were determined by direct sequence at last.h＇s firstly reported in China that DHPLC was applied in PAH gene mutation screening analysis.Results：We totally detected 13 mutations.Four novel mutations （R176X,E280K,L367R and $349A） were firstly identified in Chinese PKU population and two of them （L367R and S349A） have not been yet described in international PAH mutation database.The most prevalent mutations were R243Q （19.8%）,Y204C （7.5%）,RlllX（3.8%） and V399V（2.8%）.There was a good correlation between clinical severity and genotype （r=0.696,P〈0.01）.Conelusion：We found 4 novel mutations which extend the spectrum of Chinese PAH mutations.The consistency between the clinical severity and the genotype supplies theoretic references for optimal dietary therapy and prognosis at clinic.DHPLC is a rapid,accurate and efficiency method for PAH gene mutation screening.
出处
《中日友好医院学报》
2005年第4期198-201,F0002,共5页
Journal of China-Japan Friendship Hospital
