A transformer of molecular beacon for sensitive and real-time detection of phosphatases with effective inhibition of the false positive signals

作者：Zhang, Chen; Su, Xin; Liang, Yuan; Zhu, Xiaocui; Song, Chen; Zhao, Meiping^*

来源：Biosensors and Bioelectronics, 2011, 28(1): 13-16.

DOI：10.1016/j.bios.2011.06.049

• Molecular beacon
• Phosphatase
• Activity
• Real-time detection
• Inhibition of false-positive signals

Molecular beacons (MBs) have shown great potential in measurement of enzyme activities. However, currently available methods for monitoring of phosphatases only use MBs as a signal reporter. Extra substrates for the phosphatases are needed to hybridize to the MB either as a primer or as a template. Moreover, few MB-based methods have been used to detect enzyme activities in real biological samples due to insufficient sensitivity or false positive interference signals caused by nonspecific nucleases. In this work, a novel type of fluorescent probe was designed and synthesized for monitoring of phosphatases by integrating the DNA substrate and the signaling structures into a single molecule. Such a new design not only significantly simplified the probing system and greatly enhanced the sensitivity, but also offered a practical way to guard against the false-positive signal problems in the application to real samples. The unique design of the assay format should be widely applicable to many other enzymatic assays using oligonucleotide fluorescent probes.

• 出版日期2011-10-15
• 单位北京大学