A method for detecting microRNA based on thermophilic TtAgo, which activates mesophilic enzyme digestion activity using cofactors.
By activating the thermophilic TtAgo protein with mesophilic enzymatic cleavage activity through cofactor activation, and combining it with dCTP and TthSSB proteins, the ERCB-TtAgo method was developed. This method solves the problems of low sensitivity and complex operation in existing microRNA detection technologies, enabling rapid and simple multiplex detection, and is suitable for early tumor diagnosis.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- THE SECOND AFFILIATED HOSPITAL OF CHONGQING MEDICAL UNIV
- Filing Date
- 2026-05-27
- Publication Date
- 2026-06-30
AI Technical Summary
Existing microRNA detection technologies suffer from problems such as low sensitivity, complex operation, high cost, and insufficient multiplex detection capabilities, making it difficult to achieve rapid and convenient multiplex detection and limiting their application in early tumor diagnosis.
By activating the thermophilic TtAgo protein with mesophilic enzymatic cleavage activity through cofactors, and combining it with dCTP and TthSSB proteins, an ERCB-TtAgo method was developed to achieve isothermal multiplex analysis under mesophilic conditions. The synergistic effect of TtAgo and TthSSB is utilized to activate the cleavage activity of TtAgo, resulting in exponential accumulation of fluorescence signals.
TtAgo achieves highly sensitive and rapid multiplex microRNA detection under mesophilic conditions, with performance comparable to RT-qPCR. This expands the temperature range of TtAgo, simplifies the operation process, and reduces detection costs.
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