Convenient ATP (adenosine triphosphate) content detecting method adopting pressure-sensitive paint as signal reading tool

A pressure-sensitive coating and signal readout technology, which is applied in the field of analytical chemistry, can solve the problems of inconvenient portability, complicated operation of instruments and equipment, and inability to realize on-site detection, and achieve the effects of reducing detection costs, simple preparation process, and accurate quantitative results

Inactive Publication Date: 2016-09-21
FUZHOU UNIV
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  • Abstract
  • Description
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Problems solved by technology

However, a common disadvantage of these sensors is that they need to use laboratory equipment, which are complex to operate, inconvenient to carry, and unable to achieve on-site detection

Method used

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  • Convenient ATP (adenosine triphosphate) content detecting method adopting pressure-sensitive paint as signal reading tool

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Embodiment 1

[0052] The following examples illustrate the operation process of applying the method of the present invention to the detection of ATP concentration in human serum samples in conjunction with the accompanying drawings:

[0053] 1. Apply 0.2 mL of PtTFPP toluene solution with a concentration of 0.05 mg / mL to the inner wall of a 2 mL PE centrifuge tube and dry to obtain the signal readout tool.

[0054] 2. Synthesis of DNA compounds modified with silica spheres. References (Hermanson, G. T. Bioconjugate Techniques; Elsevier: London, 2008). Specific steps are as follows:

[0055] A. Mix 30 μmol / L DNA modified with sulfhydryl groups and 60 μmol / L TCEP in phosphate buffer solution, and incubate at room temperature for 1 hour;

[0056] B. Mix 1.00 mg of silica spheres embedded with nano-platinum and 0.5 mg of Sulfo-SMCC in a buffer solution (0.01 mol / LPBS, PH=7.3, 0.1 mol / L NaCl) for 5 minutes, and place on a shaker at room temperature Shake for 1 hour, then centrifuge the mixtur...

Embodiment 2

[0064] 1. The detection of ATP concentration in human serum, the specific steps are as follows:

[0065] Human serum was added to the mixture of DNA compounds modified with silica spheres and DNA compounds decorated on magnetic beads. After incubating at room temperature for 1.5 hours, the mixture was separated with a magnet, washed three times with phosphate buffer solution, and added 10 μL of this buffer. Finally, add the mixed solution and 300 μL of 2.0 mol / L hydrogen peroxide solution into a centrifuge tube (500 μL) at the same time, and place the centrifuge tube in the centrifuge tube whose inner wall is coated with pressure-sensitive paint in step 1 React for 15 minutes, and finally put the centrifuge tube coated with pressure-sensitive paint into a fluorescence spectrophotometer for detection. Record the data and calculate the ATP content in the human serum sample.

[0066] 2, the specificity of the method of the present invention to ATP detection, concrete steps are ...

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Abstract

The invention discloses a convenient ATP (adenosine triphosphate) content detecting method adopting pressure-sensitive paint as a signal reading tool. Two fragments of DNA capable of forming an ATP aptamer are modified with a silica sphere with embedded nano platinum particles and a magnetic bead with embedded nano platinum particles respectively. In the presence of ATP, a sandwich structure can be formed due to the specific recognition and combination of the two fragments of DNA and ATP, and accordingly, nano platinum is fixed on the magnetic bead; the magnetic bead is separated from a solution, the cleaned magnetic bead is subjected to a reaction with a hydrogen peroxide solution in a closed container, oxygen partial pressure can be increased through accumulation of oxygen generated by catalysis of hydrogen peroxide, and the concentration of ATP is indirectly detected according to fluorescence quenching degrees of different oxygen partial pressure to the pressure-sensitive paint. The technology is successfully applied to the detection of ATP concentration in human body serum; according to the detection, a fluorospectrophotometer is adopted as a detecting means, and naked eyes can also be directly used for colorimetric detection; the method has the advantages of being low in detection cost, easy to operate, accurate in quantitative result, capable of realizing instant detection, convenient and the like.

Description

technical field [0001] The invention relates to a portable method for detecting ATP content by using pressure-sensitive paint as a signal readout, belonging to the field of analytical chemistry. technical background [0002] Portable point-of-care detectors have attracted widespread attention due to their small size, low detection cost, simple operation, accurate quantitative results, and real-time detection, and have been developed and applied accordingly. Compared with traditional analysis and detection methods, portable instant detectors can get rid of the constraints of time, space, equipment cost and operation skills in the detection and analysis process to a certain extent, and can complete low-cost, timely, simple and accurate analysis and detection. It is an analytical technique that does not have too high requirements for experimental testing conditions and operators. Common portable real-time detectors include handheld real-time detectors such as thermometers, hyg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/29G01N21/64
CPCG01N21/29G01N21/6486
Inventor 林振宇李荣杰杨伟强郭隆华邱彬
Owner FUZHOU UNIV
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