Genotyping of the G1138A mutation of the FGFR3 gene in patients with achondroplasia using high-resolution melting analysis.
OBJECTIVE
The fibroblast growth factor receptor 3 gene (FGFR3) plays a critical role in cartilage growth-plate differentiation and bony development. It has been shown that 97% of patients with achondroplasia have a G to A transition mutation at position 1138 (c.1138 G>A) of codon 380 of the FGFR3 gene.


METHODS
Exon 8 of the FGFR3 gene was analyzed in 40 patients with achondroplasia, as well as in 50 control individuals for the presence of the c.1138G>A variant using melting curve analysis with a high-resolution melting instrument (HR-1).


RESULTS
The high-resolution melting curve analysis successfully genotyped the c.1138G>A mutation in exon 8 of the FGFR3 gene in all 40 patients with achondroplasia without the need of further assays. The technique had a sensitivity and specificity of 100%.


CONCLUSIONS
High-resolution melting analysis is a simple, rapid, and sensitive one tube assay for genotyping the FGFR3 gene. The technique is a low cost high-throughput FGFR3 screening assay.
Hung
Chia-Cheng
CC
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei, Taiwan.
Lee
Chien-Nan
CN
Chang
Chien-Hui
CH
Jong
Yuh-Jyh
YJ
Chen
Chih-Ping
CP
Hsieh
Wu-Shiun
WS
Su
Yi-Ning
YN
Lin
Win-Li
WL
D000130:Achondroplasia / Q000235:genetics*; D002356:Cartilage / Q000254:growth & development; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis; D051518:Fibroblast Growth Factor 3 / Q000235:genetics; D020180:Heteroduplex Analysis / Q000379:methods; D006801:Humans; D009691:Nucleic Acid Denaturation / Q000235:genetics; D017354:Point Mutation / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D051498:Receptor, Fibroblast Growth Factor, Type 3 / Q000235:genetics*; D012680:Sensitivity and Specificity; D013624:Taiwan; D044366:Transition Temperature
D051518:Fibroblast Growth Factor 3; D051498:Receptor, Fibroblast Growth Factor, Type 3