Ji Chun-yan, Sun Li-li, Cao Li-hua, Hu Yu, Huang Hong, Wang Shu-sen, Luo Yang
The Research Center for Medical Genomics, China Medical University, Shenyang, Liaoning, People's Republic of China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2011 Dec;28(6):649-53. doi: 10.3760/cma.j.issn.1003-9406.2011.06.011.
To explore the value of combining high resolution melting (HRM) with multiplex ligation-dependent probe amplification (MLPA) for detecting mutations underlying phenylketonuria.
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.
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.
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.
探讨高分辨率熔解曲线分析（HRM）与多重连接依赖探针扩增技术（MLPA）联合检测苯丙酮尿症相关基因突变的价值。
应用HRM技术检测26例苯丙酮尿症患者苯丙氨酸羟化酶基因（PAH）的小突变，结果经DNA测序验证；应用MLPA技术检测PAH基因的潜在缺失/重复；对其他潜在突变进行聚合酶链反应-限制性片段长度多态性（PCR-RFLP）分析。
共检测出44/52个等位基因（84.62%）存在21种突变，其中包括1个外显子4重复。21种突变类型中，19种为既往报道的突变，其余两种为新突变：c.584_585insA和IVS10+1G>T。此外，R243Q突变（25%）是中国最常见的突变类型。
本研究表明，HRM与MLPA联合应用可提高苯丙酮尿症基因突变的检出率，为PAH基因人类突变数据库增添了新信息，为临床诊断和产前咨询提供了依据。
