Wang Ke, Zhang Jin-Tao, Yun Yu-Xia, Wu Xiao-Bing, Chen A-Qun, Wang Peng, Wang Kai-Juan, Zhang Jian-Ying, Dai Li-Ping
Henan Key Laboratory of Tumor Epidemiology, Zhengzhou 450052, China.
Yi Chuan. 2011 Feb;33(2):182-8. doi: 10.3724/sp.j.1005.2011.00182.
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 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.
以人类基因MTHFR的1298位点为例，探讨两对引物对PCR（PCR-CTPP）的技术原理，并提高SNP基因分型的准确性。通过构建PCR-CTPP检测系统，在设计合适引物、优化退火温度和引物终浓度方面，比较了传统PCR-CTPP与改进型PCR-CTPP之间的可靠性。结果证明改进后的PCR-CTPP检测系统更加准确，这支持了关于传统PCR-CTPP理论缺陷的观点。大规模研究验证了改进方法的可靠性。预计这种改进技术将在医学和分子生物学领域得到广泛应用。
