HRM-非标记探针法检测IL-15基因SNPs方法的建立

To establish the high resolution melting analysis with unlabeled probes for genotyping IL-15 gene SNPs

目的建立用于检测白细胞介素-15(IL-15)基因单核苷酸多态性(SNP)位点的高分辨率熔解曲线(HRM)-非标记探针的方法。方法采用HRM-非标记探针法及小片段扩增法对80名健康儿童IL-15基因4个SNPs位点进行基因分型,采用基因测序法进行验证并同时与小片段扩增法分型结果进行比较。结果 HRM-非标记探针法与基因测序分型结果一致,准确率为100%;未加入温度内标的小片段扩增法不能区分野生纯合子和突变纯和子。结论 HRM-非标记探针法是对已知SNP位点突变研究的一种廉价、简便、准确的基因分型技术,适合于对已知的SNP位点或基因突变进行分型和检测。

Objective We were going to establish the high resolution melting analysis with unlabeled probes for genotyping IL- 15 gene SNP. Methods Genotypes of the IL-15 gene SNPs were determined in 80 unrelated Han healthy children using High resolution melting curve analysis with unlabeled probes, small amplification method,genotype results were corroborated by gene sequencing. Results The genotype result from unlabeled probes method and gene sequencing were consistent, the accuracy rate was 100% ; small amplification method without temperature inter calibration could not differentiate the wild homozygous and mutant homozygous. Conclusion High resolution melting curve analysis with unlabeled probes is a cheap,convenient,fast and accurate genotype technology for known SNP sites.