Abstract
Breast cancer is one of the most frequently diagnosed cancers in females worldwide and lacks specific biomarkers for early detection. In a previous study, we obtained a selective RNA-cleaving Fluorogenic DNAzyme (RFD) probe against MDA-MB-231 cells, typical breast cancer cells, through the systematic evolution of ligands by exponential process (SELEX). To improve the performance of this probe for actual application, we carried out a series of optimization experiments on the pH value of a reaction buffer, the type and concentration of cofactor ions, and sequence minimization. The length of the active domain of the probe reduced to 25 nt from 40 nt after optimization, which was synthesized more easily and economically. The detection limit of the optimized assay system was 2000 MDA-MB-231 cells in 30 min, which is more sensitive than the previous one (almost 5000 cells). The DNAzyme probe was also capable of distinguishing MDA-MB-231 cell specifically from 3 normal cells and 10 other tumor cells. This probe with high sensitivity, selectivity, and economic efficiency enhances the feasibility for further clinical application in breast cancer diagnosis. Herein, we developed an optimization system to produce a general strategy to establish an easy-to-use DNAzyme-based assay for other targets.
Highlights
Breast cancer, one of the most frequently diagnosed cancers, is the second leading cause of death by cancer in women [1,2]
252,710 new cases of invasive breast cancer will be diagnosed in women, and about 40,610 women will die from breast cancer in the United States in 2017
Many detection systems with high sensitivity and selectivity based on a RNA-cleaving Fluorogenic DNAzyme (RFD) probe have been reported in recent years, including detection of Escherichia coli (E. coli) [26,31], Clostridium difficile, [32], MDA-MB-231 cells [29], T47D cells [33], etc
Summary
One of the most frequently diagnosed cancers, is the second leading cause of death by cancer in women [1,2]. Many detection systems with high sensitivity and selectivity based on a RFD probe have been reported in recent years, including detection of Escherichia coli (E. coli) [26,31], Clostridium difficile, [32], MDA-MB-231 cells [29], T47D cells [33], etc. These special RFDs offer an excellent opportunity for designing biosensor-based DNAzymes, only a few studies optimized the RFD probes after in vitro selection [27,34,35,36,37]. The optimization promotes the development of a general strategy to establish an easy-to-use and DNAzyme-based assay for other targets
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