中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2011年
42期
2971-2976
,共6页
卢超霞%高峡%王金玮%张梅%朱宏文%孙婧%肖继芳%杨威%赵秀丽%齐展%张学
盧超霞%高峽%王金瑋%張梅%硃宏文%孫婧%肖繼芳%楊威%趙秀麗%齊展%張學
로초하%고협%왕금위%장매%주굉문%손청%초계방%양위%조수려%제전%장학
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
苯丙酮尿癥%苯丙氨痠羥化酶%突變譜%多重連接依賴式探針擴增
분병동뇨증%분병안산간화매%돌변보%다중련접의뢰식탐침확증
Phenylketonuria%Phenylalanine hydroxylase%Mutation%Multiplex ligationdependent probe amplifications
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-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.Pro147ku,p.Gly289Arg,p.Phe392Ser,p.Ile421Thr)和2种大片段缺失突变(- 4163_ - 406del和- 1932_+ 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％)、c.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.
目的 瞭解河北地區苯丙酮尿癥(PKU)患者苯丙氨痠羥化酶(PAH)基因突變情況,分析該地區PAH基因的突變譜及突變特點.方法 聯閤應用PAH基因全部外顯子及外顯子-內含子交界區擴增測序和多重連接依賴式探針擴增(MLPA)方法,對2007年9月至2009年7月就診的55例來自河北地區的PKU患者進行PAH基因點突變和拷貝數改變的檢測.對髮現大片段缺失的箇體,用跨越斷裂位點(Gap) -PCR-測序方法確定斷裂點精確位置.結果 110箇PAH等位基因中檢齣突變等位基因108箇,突變檢齣率為98.2％.共髮現PAH基因突變41種,涉及11箇外顯子,包括錯義突變24種、無義突變7種、剪接位點突變7種、小的缺失1種和大片段缺失2種,其中4種錯義突變(p.Pro147ku,p.Gly289Arg,p.Phe392Ser,p.Ile421Thr)和2種大片段缺失突變(- 4163_ - 406del和- 1932_+ 3402del)為國際首次報道.55例患者中以下3種突變所佔比例最高:p.Arg243Gln(12.7％)、c.611A ＞G(11.8％)和c.1197A ＞T(9.1％).結論 中國河北地區PKU患者中PAH基因的突變分佈廣汎;突變類型大部分為單堿基替換,但是大片段缺失也佔一定比例.
목적 료해하북지구분병동뇨증(PKU)환자분병안산간화매(PAH)기인돌변정황,분석해지구PAH기인적돌변보급돌변특점.방법 연합응용PAH기인전부외현자급외현자-내함자교계구확증측서화다중련접의뢰식탐침확증(MLPA)방법,대2007년9월지2009년7월취진적55례래자하북지구적PKU환자진행PAH기인점돌변화고패수개변적검측.대발현대편단결실적개체,용과월단렬위점(Gap) -PCR-측서방법학정단렬점정학위치.결과 110개PAH등위기인중검출돌변등위기인108개,돌변검출솔위98.2％.공발현PAH기인돌변41충,섭급11개외현자,포괄착의돌변24충、무의돌변7충、전접위점돌변7충、소적결실1충화대편단결실2충,기중4충착의돌변(p.Pro147ku,p.Gly289Arg,p.Phe392Ser,p.Ile421Thr)화2충대편단결실돌변(- 4163_ - 406del화- 1932_+ 3402del)위국제수차보도.55례환자중이하3충돌변소점비례최고:p.Arg243Gln(12.7％)、c.611A ＞G(11.8％)화c.1197A ＞T(9.1％).결론 중국하북지구PKU환자중PAH기인적돌변분포엄범;돌변류형대부분위단감기체환,단시대편단결실야점일정비례.
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.11e421Thr) 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.
