应用等位基因特异性扩增检测经典型苯丙酮尿症PAH基因第6外显子基因突变

Study on gene mutations in exon 6 of the phenylalanine hydroxylase(PAH) with classical phenylketonuria by the specific primers of allele specific amplification

目的建立快速简便、特异灵敏的鉴定PAH基因第6外显子c.611A>G突变热点的基因诊断方法。方法针对突变频率较高的PAH基因第6外显子的突变热点c.611A>G,设计等位基因特异性扩增(amplification refractory mutationsystem,ARMS)的特异引物,对山西省已经临床确诊的70例经典型PKU患儿、c.611A>G突变患儿的家长及50例正常儿童进行第6外显子扩增,扩增产物测序验证。结果在受检的山西省患儿中,PAH基因第6外显子c.611A>G突变位点ARMS检测结果和测序结果完全相符。结论 ARMS技术操作简便,重复性和稳定性好,可作为PAH基因热点突变的快速灵敏检测方法。

Objective： Establishment of the simple,rapid,specific and sensitive genetic diagnosis methods： for c.611AG of exon 6 of PAH.Methods： For the hot mutation spot c.611AG,we designed the specific primers of amplification refractory mutation system（ARMS）,70 patients with phynelketonuria,patients with their children mutationed in c.611AG,and 50 normal children were include in the study.Exon 6 amplified products were sequenced.Results： The result of mutation spot of c.611AG in exon 6 with the ARMS test are completely consistent with the DNA sequencing result.Conclusion： The method of ARMS is simple,repeatable and stable,and can be used as the method to identified the mutation hot spots of PAH.