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目的探讨高分辨熔解曲线分析法用于FGFR3基因p.R248C突变检测的可行性。方法选用经直接测序确诊基因型为p.R248C的致死性软骨发育不良Ⅰ型(type 1 thanatophoric dysplasia,TDⅠ)样本10例和30例正常对照样本,PCR扩增FGFR3基因第七外显子中包含c.742C>T(p.R248C)位点的区域。应用LightScanner 96(Idaho Technology)对PCR扩增产物进行高分辨熔解曲线分析。结果应用高分辨熔解曲线分析法成功检测出10例FGFR3基因p.R248C杂合突变。结论高分辨熔解曲线分析法是1种具有简单、高效、廉价和高通量等优点的新的分子遗传学分析法,能对FGFR3基因的p.R248C突变进行快速基因诊断。
Objective To investigate the feasibility of high-resolution melting curve analysis for the detection of p.R248C mutation in FGFR3 gene. Methods 10 cases of type Ⅰ thanatophoric dysplasia (TD Ⅰ) with genotype p.R248C and 30 normal controls were selected by direct sequencing. The seventh exon of FGFR3 gene was amplified by PCR c.742C> T (p.R248C) locus. The PCR amplification products were subjected to high resolution melting curve analysis using the LightScanner 96 (Idaho Technology). Results 10 cases of p.R248C heterozygous mutation of FGFR3 gene were successfully detected by high resolution melting curve analysis. Conclusion High-resolution melting curve analysis is a new molecular genetic analysis method which is simple, efficient, inexpensive and high-throughput. It can rapidly diagnose the p.R248C mutation of FGFR3 gene.