Furthermore, based on this polymerase/nicking endonuclease DNA machine and multiple primer-like RCAs, we designed a one-step rapid fluorescent biosensing method for ultrasensitive detection of the BCR-ABL1 fusion gene (Fig. 5D). In the strategy, the BCR-ABL1 fusion gene can be specifically identified by using dual probes to form a three-way junction structure, and then a large number of triggers are generated by the SDA reaction, which initiates the downstream nick-mediated RCA reaction. Introducing two nick endonuclease recognition sites in a circular DNA template, allowing RCA to occur in a multiple primer-like manner, the developed method yielded a broad linear response from 10fM to 1nM with a low detection limit of 5.52 fM.
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