CAJ | 학술논문

目的联合应用高分辨率熔解曲线(high resolution melting,HRM)和多重连接依赖探针扩增(multiplex ligation-dependent probe amplification,MLPA)技术检测苯丙酮尿症(phenylketonuria,PKU)患者基因突变,并探讨两种技术联合运用对于突变筛查的价值.方法采用HRM和MLPA技术对26例临床诊断为PKU的患者苯丙氨酸羟化酶基因(phenylalanine hydroxylase,PAH)进行检测,并通过测序验证；针对未报道的基因改变进行聚合酶链反应-限制性片段长度多态性分析.结果在52个等位基因中检测到44个突变等位基因(共21种不同突变),包括第4外显子拷贝数的增加,总检出率达84.62％(44/52),其中c.584_585insA和IVS10+ 1G＞T突变在国内外未见报道.此外,R243Q(25％)突变为中国最常见的突变种类.结论应用HRM和MLPA技术相对提高了PAH基因突变筛查的检出率,检测结果丰富了基因突变数据库,并为临床诊断与产前诊断提供实验室依据.
목적 연합응용고분변솔용해곡선(high resolution melting,HRM)화다중련접의뢰탐침확증(multiplex ligation-dependent probe amplification,MLPA)기술검측분병동뇨증(phenylketonuria,PKU)환자기인돌변,병탐토량충기술연합운용대우돌변사사적개치.방법 채용HRM화MLPA기술대26례림상진단위PKU적환자분병안산간화매기인(phenylalanine hydroxylase,PAH)진행검측,병통과측서험증；침대미보도적기인개변진행취합매련반응-한제성편단장도다태성분석.결과 재52개등위기인중검측도44개돌변등위기인(공21충불동돌변),포괄제4외현자고패수적증가,총검출솔체84.62％(44/52),기중c.584_585insA화IVS10+ 1G＞T돌변재국내외미견보도.차외,R243Q(25％)돌변위중국최상견적돌변충류.결론 응용HRM화MLPA기술상대제고료PAH기인돌변사사적검출솔,검측결과봉부료기인돌변수거고,병위림상진단여산전진단제공실험실의거.
Objective To explore the value of combining high resolution melting (HRM) with multiplex ligation-dependent probe amplification ( MLPA ) for detecting mutations underlying phenylketonuria.Methods HRM was used for detecting small mutations in phenylalanine hydroxylase gene (PAH) of 26 phenylketonuria patients.The results were verified with DNA sequencing.MLPA was used for detecting potential deletions/duplications in the PAH gene. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was performed for additional potential mutations.Results A total of 21 mutations were found in 44/52 alleles (84.62％),which included a dupEx4.Among the 21 types of mutation,19 were reported previously,and the remaining two were novel mutations:c.584_585insA and IVS10+1G＞T.In addition,the mutation of R243Q (25％) was the most common type in China.Conclusion The study showed that the combination of HRM and MLPA could increase the detection rate for mutation in PKU.The study has added new information to the human mutation database of PAH and provided a basis for clinical diagnosis and prenatal counseling.