Wang Sihua, Ding Mingcui, Duan Xiaoran, Wang Tuanwei, Feng Xiaolei, Wang Pengpeng, Yao Wu, Wu Yongjun, Yan Zhen, Feng Feifei, Yu Songcheng, Wang Wei
Department of Occupational Health, College of Public Health, Zhengzhou University, Zhengzhou, Henan, China.
Department of Occupational Health, Henan Institute of Occupational Health, Zhengzhou, Henan, China.
Ann Clin Lab Sci. 2017 Sep;47(5):546-550.
It has been shown that the single nucleotide polymorphism (SNP) of the rs2735940 site in the human telomerase reverse transcriptase () gene is associated with increased cancer risk. The traditional method to detect SNP genotypes is polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). However, there is a limitation to utilizing PCR-RFLP due to a lack of proper restriction enzyme sites at many polymorphic loci. This study used an improved PCR-RFLP method with a mismatched base for detection of the SNP rs2735940. A new restriction enzyme cutting site was created by created restriction site PCR (CRS-PCR), and in addition, the restriction enzyme I for CRS-PCR was cheaper than other enzymes. We used this novel assay to determine the allele frequencies in 552 healthy Chinese Han individuals, and found the allele frequencies to be 63% for allele and 37% for allele In summary, the modified PCR-RFLP can be used to detect the SNP of rs2735940 with low cost and high efficiency.
研究表明，人类端粒酶逆转录酶（）基因中rs2735940位点的单核苷酸多态性（SNP）与癌症风险增加有关。检测SNP基因型的传统方法是聚合酶链反应-限制性片段长度多态性（PCR-RFLP）。然而，由于许多多态性位点缺乏合适的限制性酶切位点，利用PCR-RFLP存在局限性。本研究采用一种改进的带有错配碱基的PCR-RFLP方法来检测SNP rs2735940。通过创建限制性位点PCR（CRS-PCR）创建了一个新的限制性酶切位点，此外，用于CRS-PCR的限制性酶I比其他酶更便宜。我们使用这种新方法测定了552名健康中国汉族个体的等位基因频率，发现等位基因频率为63%，等位基因频率为37%。总之，改良的PCR-RFLP可用于低成本、高效率地检测rs2735940的SNP。
