Rapid detection of hot spot mutations of FGFR3 gene with PCR-high resolution melting assay.
- Author:
Shan LI
1
;
Han WANG
;
Hua SU
;
Jinsong GAO
;
Xiuli ZHAO
Author Information
- Publication Type:Journal Article
- MeSH: DNA Mutational Analysis; methods; Female; Humans; Male; Mutation; genetics; Polymerase Chain Reaction; methods; Receptor, Fibroblast Growth Factor, Type 3; genetics; Transition Temperature
- From: Chinese Journal of Medical Genetics 2017;34(4):494-498
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo identify the causative mutations in five individuals affected with dyschondroplasia and develop an efficient procedure for detecting hot spot mutations of the FGFR3 gene.
METHODSGenomic 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.
RESULTSA 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.
CONCLUSIONThe 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.
