Portable method for detecting content of adenosine triphosphate (ATP)

An adenosine triphosphate, portable detection technology, applied in the field of portable detection of adenosine triphosphate content, can solve the problems of inconvenience to carry, complicated operation of instruments and equipment, inability to realize on-site detection, etc., and achieves the effects of good activity and stability, non-toxic cost, and low cost.

Inactive Publication Date: 2013-10-09
FUZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, a common disadvantage of these sensors is that they need to use laboratory equipment

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  • Portable method for detecting content of adenosine triphosphate (ATP)

Examples

Experimental program
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Example Embodiment

[0030] Example 1

[0031] The following embodiments are combined with the drawings to illustrate the operation process of applying the method of the present invention to the detection of ATP concentration in human serum samples:

[0032] 1. Synthesis of DNA compounds modified with sucrose invertase. References (Hermanson, G. T. Bioconjugate Techniques; Elsevier: London, 2008). Specific steps are as follows:

[0033] A. Mix 30 μM DNA with sulfhydryl groups and 60 μM TCEP in PBS buffer solution, and incubate at room temperature for 1 hour;

[0034] B. Mix 10.0 mg / mL sucrose invertase and 1 mg Sulfo-SMCC in a buffer solution (0.01 M PBS, pH=7.3, 0.1 M NaCl) for 5 minutes, place on a shaker at room temperature and shake for 1 hour, and then mix the mixture Centrifugal separation, take the supernatant;

[0035] C. Mix the substances prepared by A and B, and react for 48 hours at room temperature.

[0036] 2. Preparation of magnetic beads modified with sucrose invertase and detection of ATP...

Example Embodiment

[0041] Example 2

[0042] 1. To detect the concentration of ATP in human serum, the specific steps are as follows: add a dilution of 10 to 4.0 uM of two modified DNA mixtures 3 Incubate the human serum sample at room temperature for (1.5±0.05) hours, then separate the mixture with a magnet, wash the magnetic beads three times with 50 uL buffer solution and add 10 uL buffer solution, then add 20 uL 1 M sucrose solution, and react at 45℃ In 20 minutes, pipette 2.5 μL to measure with a blood glucose meter, and record the data. The test results show that the ATP concentration in human serum samples is approximately 1.81×10 -10 M.

[0043] 2. The specific steps of the method of the present invention for ATP detection are as follows: In order to detect the specificity of the method of the present invention, the ATP used in Examples 1-2 is replaced with other interfering substances, which are UTP respectively. , GTP and CTP, the concentration of ATP is 1×10 -12 The concentration of M, ...

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Abstract

The invention discloses a portable method for detecting content of adenosine triphosphate (ATP). The method comprises the following steps of: respectively modifying a magnetic bead and invertase on an ATP aptamer which is split into two sections, performing specific identification and combination on the ATP aptamer which is split into two sections and a target ATP so as to form a sandwich structure, so that the aptamer modified with the invertase is fixed on the magnetic bead; separating the magnetic bead from the solution, dissolving the cleaned magnetic bead in a sucrose solution for carrying out an enzymatic hydrolysis reaction, measuring the reaction solution subjected to enzymatic hydrolysis by employing a glucometer, and indirectly detecting the concentration of ATP. The method is successfully applied to detection of the ATP concentration in a human serum sample, and the method has the advantages of simple operation, low cost, high sensitivity, high specificity and the like.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and in particular relates to a portable method for detecting the content of adenosine triphosphate. Background technique [0002] Adenosine triphosphate (ATP) is the direct and main source of energy required for all life activities of tissue cells in the body. It can store and transfer chemical energy, participate in the synthesis of proteins, fats, sugars and nucleotides, and can promote the repair of various cells in the body and regeneration, enhance cell metabolism activity, and have strong pertinence for the treatment of various diseases. Therefore, quantitative detection of ATP content has important practical significance for scientific research and clinical diagnosis. [0003] Aptamers are a class of single-stranded DNA or RNA oligonuclei that can bind to target molecules with high affinity and recognize target molecules with high specificity through in vitro screening by exponential e...

Claims

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Application Information

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IPC IPC(8): G01N27/26
Inventor 林振宇周津郭隆华邱彬陈国南
Owner FUZHOU UNIVERSITY
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