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Ligation-dependent rolling circle amplification method for ATP determination with high selectivity and sensitivity.

Mingjian Chen1,2,3, Yang Li4, Peng Li4

  • 1NHC Key Laboratory of Carcinogenesis and Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute and School of Basic Medical Science, Central South University, Changsha 410078, P. R. China. wuxu1028@csu.edu.cn.

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A new method reliably detects adenosine triphosphate (ATP) using a specific ligation reaction and dual signal amplification. This sensitive and selective approach offers a convenient alternative for ATP detection in various applications.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Adenosine triphosphate (ATP) is crucial in clinical diagnosis, biomedical engineering, and food chemistry.
  • Current ATP detection methods often require expensive instruments or exhibit limited sensitivity and selectivity.
  • There is a need for efficient, reliable, and cost-effective ATP sensing technologies.

Purpose of the Study:

  • To develop a highly sensitive and selective method for detecting adenosine triphosphate (ATP).
  • To overcome the limitations of existing ATP sensing techniques, such as high cost and low performance.
  • To provide a versatile platform for biomolecule detection applicable to real-world samples.

Main Methods:

  • Integration of an ATP-dependent ligation reaction with dual-stage signal amplification.
  • Utilizing rolling circle amplification (RCA) for initial signal amplification.
  • Employing DNAzymes for catalytic cleavage and fluorescence signal generation from molecular beacons (MBs).

Main Results:

  • Achieved a low detection limit of 35 pM for ATP.
  • Demonstrated a linear detection range from 0.05 nM to 200 nM.
  • Successfully discriminated ATP from its analogues and validated performance in spiked human serum albumin (HSA) samples.

Conclusions:

  • The proposed method offers high sensitivity and selectivity for ATP detection.
  • The strategy is wash-free, isothermal, and features a simple probe design.
  • This approach provides a novel and effective platform for determining ATP and other biomolecules.