CAJ | 학술논문

目的探讨宁夏地区苯丙酮尿症(phenylketonuria,PKU)患儿苯丙氨酸羟化酶(phenylalanine hydroxylase,PAH)基因突变特点. 方法 2010年1月至2013年6月,在宁夏回族自治区妇幼保健院诊断为PKU的73例患儿纳入研究,在同期PKU筛查结果正常的新生儿中随机选取100例作为对照组.取静脉血,采用聚合酶链反应技术扩增PAH基因,并采用直接测序法进行基因测序.检索国内外相关文献及数据库判断发现的基因突变是否为新突变.采集患儿父母血标本进行突变测序,证实新突变的来源.同时对对照组进行基因测序,以证实该突变是否为新突变.采用x2检验进行统计学分析. 结果 73例患儿146个PAH等位基因中突变基因检出率为79.5％(116/146),包含37种突变类型,分布于除2和13以外的11个外显子及部分外显子的旁侧内含子上.其中错义突变22种(59.5％,22/37)、无义突变6种(16.2％,6/37)、剪切位点突变6种(16.2％,6/37)和缺失突变3种(8.1％,3/37).最常见的基因突变类型是p.R243Q,占17.1％(25/146),其次是EX6-96A ＞G(6.8％,10/146)、p.R241C(6.2％,9/146)、p.R413P(5.5％,8/146)、p.RlllX (4.8％,7/146)和IVS4-1G＞A(4.8％,7/146).回、汉族患儿最常见的基因突变类型均是p.R243Q[分别为15.9％(13/82)和18.8％ (12/64)].p.R241C检出率回族高于汉族[9.8％(8/82)与1.6％(1/64),x2=4.17,P=0.04].检索及对比发现,p.Q304K、p.H107R、p.F392I和p.N223I可能是未报道的PAH基因突变类型. 结论宁夏地区PKU患儿PAH基因突变具有民族地区基因突变的多样性和复杂性.
목적 탐토저하지구분병동뇨증(phenylketonuria,PKU)환인분병안산간화매(phenylalanine hydroxylase,PAH)기인돌변특점. 방법 2010년1월지2013년6월,재저하회족자치구부유보건원진단위PKU적73례환인납입연구,재동기PKU사사결과정상적신생인중수궤선취100례작위대조조.취정맥혈,채용취합매련반응기술확증PAH기인,병채용직접측서법진행기인측서.검색국내외상관문헌급수거고판단발현적기인돌변시부위신돌변.채집환인부모혈표본진행돌변측서,증실신돌변적래원.동시대대조조진행기인측서,이증실해돌변시부위신돌변.채용x2검험진행통계학분석. 결과 73례환인146개PAH등위기인중돌변기인검출솔위79.5％(116/146),포함37충돌변류형,분포우제2화13이외적11개외현자급부분외현자적방측내함자상.기중착의돌변22충(59.5％,22/37)、무의돌변6충(16.2％,6/37)、전절위점돌변6충(16.2％,6/37)화결실돌변3충(8.1％,3/37).최상견적기인돌변류형시p.R243Q,점17.1％(25/146),기차시EX6-96A ＞G(6.8％,10/146)、p.R241C(6.2％,9/146)、p.R413P(5.5％,8/146)、p.RlllX (4.8％,7/146)화IVS4-1G＞A(4.8％,7/146).회、한족환인최상견적기인돌변류형균시p.R243Q[분별위15.9％(13/82)화18.8％ (12/64)].p.R241C검출솔회족고우한족[9.8％(8/82)여1.6％(1/64),x2=4.17,P=0.04].검색급대비발현,p.Q304K、p.H107R、p.F392I화p.N223I가능시미보도적PAH기인돌변류형. 결론 저하지구PKU환인PAH기인돌변구유민족지구기인돌변적다양성화복잡성.
Objective To characterize the mutation of phenylalanine hydroxylase (PAH) gene in patients with phenylketonuria(PKU) in Ningxia area,China.Methods Seventy-three children diagnosed with PKU at the Child and Maternal Healthcare Hospital of Ningxia Hui Autonomous Region between January 2010 and June 2013,and 100 non-PKU children randomly chosen from children with normal results in PKU screening were enrolled in the study.Venous blood was collected and the PAH gene sequence was determined by direct DNA sequencing after amplification with the polymerase chain reaction technique.The new gene mutations were defined based on the national and international literature search and databases.The source of the newly discovered mutations was also measured by examining and sequencing the blood samples of their parents.The Chi-square test was used for statistical analysis.Results Among 146 alleles of the 73 PKU children,the detection rate of mutation of PAH gene was 79.5％ (116/146),including 37 types of mutations occurring in 11 exons other than exon 2 and exon 13.The 37 different mutations included 22 missense mutations (59.5％,22/37),six nonsense mutations(16.2％,6/37),six splice site mutations(16.2％,6/37) and three deletion mutations(8.1％,3/37).p.R243Q(17.1％,25/146),EX6-96A ＞ G (6.8％,10/146),p.R241C(6.2％,9/146),p.R413P (5.5％,8/146),p.Rl11X(4.8％,7/146) and IVS4-1G ＞ A(4.8％,7/146) were found to have a higher mutation frequency.Meanwhile,p.R243Q was the most common mutation among Han and Hui ethnic groups with a frequency of 18.8％(12/64) and 15.9％ (13/82),respectively.In contrast,p.R241C showed a significant higher frequency in the Hui group [9.8％(8/82) vs 1.6％(1/64),x2=4.17,P=0.04].Four new mutations of PAH genes,including p.Q304K,p.H107R,p.F392I and p.N223I,were discovered after literature search and comparative studies.Conclusions PAH gene mutations in children with PKU in Ningxia area are unique and are characterized by the diversity and complexity of mutation occurrence in this ethnic region.