PCR-CTPP:一种基于错配技术的SNP分型方法的改良

摘要

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 AfTHFR as an example, the reliability between conventional 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 conventional 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.%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.