[Rapid detection of hot spot mutations of FGFR3 gene with PCR-high resolution melting assay]

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.
[Article in Chinese]

Abstract

Objective: 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.

Methods: 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.

Results: 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.

Conclusion: 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.

MeSH terms

  • DNA Mutational Analysis / methods
  • Female
  • Humans
  • Male
  • Mutation / genetics*
  • Polymerase Chain Reaction / methods
  • Receptor, Fibroblast Growth Factor, Type 3 / genetics*
  • Transition Temperature

Substances

  • FGFR3 protein, human
  • Receptor, Fibroblast Growth Factor, Type 3