变性高效液相色谱法筛选鼻咽癌相关基因单核苷酸多态性

Screening of single nucleotide polymorphisms in nasopharyngeal carcinoma associated genes by denaturing high-performance liquid chromatography

目的 利用变性高效液相色谱法（DHPLC）结合测序筛选和鉴定鼻咽癌基因的单核苷酸多态性（SNPs）。方法 PCR扩增30例鼻咽癌患者和28例正常对照者6p21.3区域基因PPP1R11、PP1R10、FLOT1、KIAA0170的4个外显子与1个内含子片断,采用DHPLC技术对扩增片断进行基因变异检测,将不同的类型PCR片断进行全序列测定并与参考序列对照分析。结果 在5个片断中初步鉴定出4个未见报道的新SNP位点,验证了4个已知的SNPs位点和基因型。结论 DHPLC预测结合全序列测序是一种高效、经济、简便、可靠的SNP筛选方法。

Objective To screen single nucleotide polymorphisms (SNPs) of 4 human genes at 6p21.3 by way of denaturing high-performance liquid chromatography (DHPLC). Method Four exons and 1 intron fragments in the PPP1R11, PPP1R10, FLOTl and KIAA0170 genes at 6p21.3 isolated from the blood samples from 30 patients with nasopharyngeal carcinoma (NPC) and 28 healthy subjects were amplified by PCR, and the products analyzed by DHPLC. Some fragments of interest were sequenced and compared with the sequences available in National Center for Biotechnology Information (NCBI) database. Results In the 5 fragments, 4 new SNPs were identified and 4 known SNP loci and genotypes were confirmed. Conclusion DHPLC is an effective, economical, and simple method with reliability for SNPs screening.