PCR-CTPP:一种基于错配技术的SNP分型方法的改良  

Improvement on PCR-CTPP:a SNP genotyping approach based on mismatch technique

作　　者：王柯[1,2] 章金涛[2,3] 运玉霞[4,1] 吴晓冰[4,1] 陈阿群[4,1] 王鹏[4,1] 王凯娟[4,1] 张建营[4,1] 代丽萍[4,1] 

机构地区：[1]河南省肿瘤流行病学重点实验室,郑州450052 [2]郑州大学生物工程系,郑州450001 [3]郑州大学实验动物中心,郑州450052 [4]郑州大学公共卫生学院流行病与卫生统计学系,郑州450001

出　　处：《遗传》2011年第2期182-188,共7页Hereditas（Beijing)

基　　金：国家自然科学基金项目(编号:30872181;81072360);河南省高校科技创新人才支持计划(编号:2010HASTIT027);河南医学科技攻关项目(编号:200903009)资助

摘　　要：为探讨两对交叉引物PCR(PCR-CTPP)技术的原理并提高SNP分型的准确性,以人类MTHFR基因1298位点突变为例,通过设计合理的引物、优化引物终浓度及复性温度等重建PCR-CTPP检测系统,对比常规PCR-CTPP和改良的PCR-CTPP检测系统的可靠性。结果表明,改良的PCR-CTPP检测系统更加准确,支持了常规方法存在理论缺陷的观点;批量鉴定结果进一步验证了改良方法的可靠性。该技术有望在医学和分子生物学等领域广泛应用。To explore the technique principle of PCR with confronting two-pair primers （PCR-CTPP） and improve the accuracy of SNP genotyping by taking the 1298 locus of human gene MTHFR as an example, the reliability between con- ventional PCR-CTPP and improved PCR-CTPP was compared using reconstructed PCR-CTPP detecting system in terms of designing appropriate primers and optimizing annealing temperature and the final concentration of primers. The improved PCR-CTPP detection system proved to be more accurate, which supported the viewpoint on the theoretical defects of con- ventional PCR-CTPP. The large-scale study verified the reliability of the improved method. It is expected that this improved technique would be widely used in the field of medicine and molecular biology.

关 键 词：两对交叉引物PCR 四引物扩增受阻突变体系PCR SNP分型 人为错配 亚甲基四氢叶酸还原酶 

分 类 号：Q78[生物学—分子生物学]