title: Rapid detection of FGFR3 gene mutation in achondroplasia by DHPLC system-coupling heteroduplex and fluorescence-enhanced primer-extension analysis.
abstract: Achondroplasia is a common form of human dwarfism with characteristically rhizomelic shortening of extremities and relative macrocephaly. It is transmitted as an autosomally dominant inheritance, and about 80% of affected individuals result from sporadic mutations without positive family histories. Achondroplasia comes from the genetic point mutations in the fibroblastic growth factor receptor 3 gene (FGFR3), which enables abnormal cartilage growth-plate differentiation and insufficient bony development. The most common genetic mutations in this receptor are G to A at position 1138 (G1138A), which result in the substitution of glycine to arginine at codon 380. Based on genetic information, molecular genetic testing can provide an exact diagnosis comparing to radiological and prenatal ultrasound evaluations. Here we introduce denaturing high-performance liquid chromatography (DHPLC) for the detection of 17 cases of achondroplasia and 120 unaffected cases. After coupling heteroduplex and fluorescence-enhanced primer-extension analysis, all affected patients with G1138A were identified successfully. In conclusion, we demonstrated that DHPLC is an efficient, accurate, and sensitive technique to detect the single gene mutation of achondroplasia in clinical applications.
authors.affiliation: Department of Medical Genetics, National Taiwan University Hospital, National Taiwan University, Taipei, Taiwan, ROC.
authors.forename: Yi-Ning
authors.initials: YN
authors.lastname: Su
authors.affiliation: Department of Obstetrics and Gynecology, National Taiwan University Hospital, College of Medicine, National Taiwan University, No.7, Chung-Shan South Road, Taipei, Taiwan, ROC.
authors.forename: Chien-Nan
authors.initials: CN
authors.lastname: Lee
authors.affiliation: Department of Obstetrics and Gynecology, National Taiwan University Hospital, College of Medicine, National Taiwan University, No.7, Chung-Shan South Road, Taipei, Taiwan, ROC.
authors.forename: Shu-Chin
authors.initials: SC
authors.lastname: Chien
authors.affiliation: Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan, ROC.
authors.forename: Chia-Cheng
authors.initials: CC
authors.lastname: Hung
authors.affiliation: Department of Medical Genetics, National Taiwan University Hospital, National Taiwan University, Taipei, Taiwan, ROC.|Department of Pediatrics, National Taiwan University Hospital, College of Medicine, National Taiwan University, Taipei, Taiwan, ROC.
authors.forename: Yin-Hsiu
authors.initials: YH
authors.lastname: Chien
authors.affiliation: Department of Obstetrics and Gynecology, National Taiwan University Hospital, College of Medicine, National Taiwan University, No.7, Chung-Shan South Road, Taipei, Taiwan, ROC. cachen@ntumc.org.
authors.forename: Chi-An
authors.initials: CA
authors.lastname: Chen
mesh_terms: D000130:Achondroplasia / Q000175:diagnosis* / Q000235:genetics; D002356:Cartilage / Q000254:growth & development; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D005453:Fluorescence; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D009691:Nucleic Acid Denaturation / Q000235:genetics; D017354:Point Mutation / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D051498:Receptor, Fibroblast Growth Factor, Type 3; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics*
chemical_list: D017931:DNA Primers; D017468:Receptors, Fibroblast Growth Factor; C496365:FGFR3 protein, human; D011505:Protein-Tyrosine Kinases; D051498:Receptor, Fibroblast Growth Factor, Type 3