飞行时间质谱法检测干扰素调节因子6单核苷酸多态性的研究

Fast and High-throughput Genotyping of SNPs of Cleft Related Gene IRF6 Using MALDI-TOF

目的研究基质辅助激光解吸电离飞行时间质谱(matrix assisted laser desorption ionization time of flightmass spectrometry,MALDI-TOF-MS)技术检测唇腭裂相关基因干扰素调节因子6(interferon regulatory factor 6,IRF6)单核苷酸多态性(single nucleotide polymorphisms,SNPs)的效果。方法样本来自201名健康志愿者,利用PCR扩增出IRF6基因含rs2235371和rs642961位点片段作为模板,并以位点特异引物进行单碱基延伸反应,产物纯化后利用MALDI-TOF-MS技术对rs2235371和rs642961的多态性进行检测。结果 201名健康志愿者IRF6基因rs2235371和rs642961位点的基因型和等位基因的分布频率与HAPMAP基因库中中国人群同一基因位点的分布频率相近。结论 MALDI-TOF-MS技术是一种高通量快速检测基因SNPs的方法。

Objective To establish a high throughput,fast and accurate method for genotyping of SNPs of cleft related gene IRF6.Methods IRF6 gene fragment including rs2235371 and rs642961 was amplified and purified as the template of the following primer extension reaction.The single base extension reaction was carried out using SNP specific extension primer.The products were purified and analyzed by MALDI-TOF.Results The method for fast and high throughput genotyping rs2235371 and rs642961 has been set up.The frequencies of rs2235371 and rs642961 in our 201 unrelated healthy Han Chinese using genotyping by MALDI-TOF are as same as reported frequencies of Han Chinese in HapMap.Conclusion MALDI-TOF is a fast,high throughput and accurate method for genotyping,and is suitable for large sample cleft related genes screening.