Weizmann Yossi, Patolsky Fernando, Katz Eugenii, Willner Itamar
Institute of Chemistry, The Hebrew University of Jerusalem, Jerusalem 91904, Israel.
Chembiochem. 2004 Jul 5;5(7):943-8. doi: 10.1002/cbic.200300820.
A highly sensitive telomerase detection method that involves amplified telomerase analysis and the use of rotating magnetic particles has been developed. Magnetic particles, functionalized with a primer (1) that is recognized by telomerase, are mixed with a nucleotide mixture that includes biotinylated-dUTP, and telomerase-induced elongation of the primers proceeds with simultaneous biotin incorporation. Avidin-Horseradish peroxidase conjugate, coupled to biotin labels, yields the biocatalytic functional particles. Mixing the resulting particles with naphthoquinone-modified magnetic particles enables the optoelectronic detection of telomerase. Attraction of the magnetic particles to an electrode, followed by rotation of the particles, causes the electrocatalytic reduction of O(2) to H(2)O(2) and HRP-catalyzed oxidation of luminol (3); this results in chemilumunescence. The intensity of the emitted light depends on the telomerase content of the sample and the rotation speed of the particles. A minimum number of 10 cancer cells could be detected.
已开发出一种高度灵敏的端粒酶检测方法,该方法涉及扩增端粒酶分析和旋转磁性颗粒的使用。用端粒酶识别的引物(1)功能化的磁性颗粒与包含生物素化-dUTP的核苷酸混合物混合,端粒酶诱导的引物延伸伴随着生物素的同时掺入。与生物素标签偶联的抗生物素蛋白-辣根过氧化物酶共轭物产生生物催化功能颗粒。将所得颗粒与萘醌修饰的磁性颗粒混合可实现端粒酶的光电检测。磁性颗粒被吸引到电极上,随后颗粒旋转,导致O(2)电催化还原为H(2)O(2)以及HRP催化鲁米诺(3)氧化;这会产生化学发光。发射光的强度取决于样品中端粒酶的含量和颗粒的旋转速度。最少可检测到10个癌细胞。