摘要
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 explore the feasibility of using high-resolution melting curve to detect the c.742C>T(p.R248C) mutation of the FGFR3 gene causing type 1 thanatophoric dysplasia. Methods A 96-well LightScanner (Idaho Technology) for high resolution melting analysis was used to genotype the c.742C>T mutation within the exon 7 of the FGFR3 gene . DNA samples including 10 prenatal samples with type 1 TD harboring the c.742 C>T variant identified previously by DNA sequencing and 30 control samples from healthy individuals were amplified with the double-stranded DNA binding dye LC Green Plus. Melting curves were analyzed as fluorescence difference plots. Results Using the high-resolution melting curve analysis,we successfully genotyped the c.742C>T (p.R248C) mutation in all of the 10 tested samples with type 1 TD. Conclusion High-resolution melting analysis is a kind of easy-to-use,rapid,and sensitive one tube assay for genotyping the c.742C>T (p.R248C)mutation which is the most common one associated with type 1 TD. The technology has the potential to reduce sequencing burden and would be suitable for mutations screening of FGFR3 gene that have been found to be associated with TD.
出处
《中国产前诊断杂志（电子版）》
2010年第2期19-23,共5页
Chinese Journal of Prenatal Diagnosis(Electronic Version)
基金
广东省科技厅一般项目(2008B080702011)
