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基于分子信标技术的CYP2C8基因139A/G单核苷酸多态性检测的初步研究

, PP. 1075-1079

Keywords: 分子信标,断接式发夹探针,单核苷酸多态性,CYPC基因

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Abstract:

目的基于分子信标技术设计一种新型的断接式发夹探针,利用该探针建立并优化检测CYP2C8基因139A/G单核苷酸多态性(singlenucleotidepolymorphism,SNP)的方法。方法首先运用生物信息学软件设计断接式发夹探针,合成目的探针和靶序列,再探索反应体系的最适温度和最适探针浓度比,最后利用该方法对CYP2C8139A/G的SNP进行检测。结果断接式发夹探针识别靶序列中SNP位点的最适杂交温度为40℃,反应体系中荧光探针与淬灭探针的最适浓度比为1∶4。在最适温度和最适探针浓度比的反应条件下,断接式发夹探针与CYP2C8基因原始靶序列反应体系的荧光强度为(7632.0±8.7)RFU,而与突变靶序列反应体系的荧光强度为(6425.7±6.1)RFU,两者差异有统计学意义(P<0.05)。结论新型的断接式发夹探针可以有效区分原始序列和突变序列,是一种可以用于检测CYP2C8基因139A/GSNP位点的新方法。

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