Li Shan, Wang Han, Su Hua, Gao Jinsong, Zhao Xiuli
Department of Medical Genetics, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences-Peking Union Medical College School of Basic Medicine, Beijing 100005, China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2017 Aug 10;34(4):494-498. doi: 10.3760/cma.j.issn.1003-9406.2017.04.006.
To identify the causative mutations in five individuals affected with dyschondroplasia and develop an efficient procedure for detecting hot spot mutations of the FGFR3 gene.
Genomic DNA was extracted from peripheral blood samples with a standard phenol/chloroform method. PCR-Sanger sequencing was used to analyze the causative mutations in the five probands. PCR-high resolution melting (HRM) was developed to detect the identified mutations.
A c.1138G>A mutation in exon 8 was found in 4 probands, while a c.1620C>G mutation was found in exon 11 of proband 5 whom had a mild phenotype. All patients were successfully distinguished from healthy controls with the PCR-HRM method. The results of HRM analysis were highly consistent with that of Sanger sequencing.
The Gly380Arg and Asn540Lys are hot spot mutations of the FGFR3 gene among patients with ACH/HCH. PCR-HRM analysis is more efficient for detecting hot spot mutations of the FGFR3 gene.
鉴定5例软骨发育异常患者的致病突变，并开发一种检测FGFR3基因热点突变的有效方法。
采用标准酚/氯仿法从外周血样本中提取基因组DNA。使用PCR-Sanger测序分析5名先证者的致病突变。开发了PCR高分辨率熔解曲线分析（HRM）来检测已鉴定的突变。
在4名先证者中发现第8外显子的c.1138G>A突变，而在表型较轻的先证者5的第11外显子中发现c.1620C>G突变。所有患者均通过PCR-HRM方法成功与健康对照区分开来。HRM分析结果与Sanger测序结果高度一致。
Gly380Arg和Asn540Lys是ACH/HCH患者中FGFR3基因的热点突变。PCR-HRM分析在检测FGFR3基因热点突变方面更有效。
