College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, PR China.
Biosens Bioelectron. 2011 Jun 15;26(10):4095-8. doi: 10.1016/j.bios.2011.03.041. Epub 2011 Apr 13.
In the present study, a chemiluminescence method for sensitive detection of human telomerase activity was developed based on the formation of G-quadruplex-hemin DNAzyme. In the presence of telomerase, the telomerase substrate (TS) primer elongated and a long single-strand DNA containing the telomere repeat units (TTAGGG)n was formed. When K(+) was introduced, the telomere repeat units could form G-quadruplex and then combined with hemin to form DNAzymes which could stimulate the generation of chemiluminescence (CL) in the presence of luminol and H(2)O(2). The amount of telomerase elongation product was controlled by the content of telomerase extracted from HeLa cells, so the amount of DNAzymes and the intensity of chemiluminescence signal were all related to the number of HeLa cells. Using this simple method, the telomerase activity extracted from 100 cultured cancer cells could be detected without the polymerase chain reaction (PCR) amplification of telomerase elongated product.
在本研究中，基于 G-四链体-血红素 DNA zyme 的形成，开发了一种用于灵敏检测人端粒酶活性的化学发光法。在端粒酶存在的情况下，端粒酶底物（TS）引物延伸，形成含有端粒重复单元（TTAGGG）n 的长单链 DNA。当引入 K(+) 时，端粒重复单元可以形成 G-四链体，然后与血红素结合形成 DNAzymes，在鲁米诺和 H(2)O(2)存在下可以刺激化学发光（CL）的产生。端粒酶延伸产物的量受从 HeLa 细胞中提取的端粒酶含量的控制，因此 DNAzymes 的量和化学发光信号的强度都与 HeLa 细胞的数量有关。使用这种简单的方法，无需端粒酶延伸产物的聚合酶链反应（PCR）扩增，即可检测 100 个培养的癌细胞中提取的端粒酶活性。
