Method for detecting enzymatic activity by using quantum dot fluorescence

A fluorescence detection and quantum dot technology, which is applied in the direction of fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of detection of enzyme disease markers that have not been reported, and achieve strong optical anti-interference ability, fast detection, sensitive qualitative detection Effect

Inactive Publication Date: 2012-05-30
TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, quantum dots have been applied to glucose (X.Y.Li, Y.L.Zhou, Z.Z.Zheng, X.L.Yue, Z.F.Dai, S.Q.Liu, Z.Y.Tang, Langmuir, 2009, 25, 6580-6586), galactose (R.Freeman, L. Bahshi, T.Finder, R.Gill, I.Willner, Chem.Commun., 2009, 764-766), nucleic acid (W.R.Algar, U.J.Krull, Anal.Chem., 2009, 81, 4113-4120) and other physiological indicators detection of enzymes, but studies on the detection of enzyme disease markers have not been reported

Method used

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  • Method for detecting enzymatic activity by using quantum dot fluorescence
  • Method for detecting enzymatic activity by using quantum dot fluorescence
  • Method for detecting enzymatic activity by using quantum dot fluorescence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Preparation of cadmium telluride quantum dots (prepared with reference to CN 200810101429.4)

[0031] (1) Add cadmium nitrate and thioglycolic acid in sequence in deionized water, so that the molar ratio of cadmium nitrate and thioglycolic acid is 1:50. After the complexation reaction is completed, add sodium hydride telluride equimolar to cadmium nitrate to obtain water-soluble quantum point precursor solution, the final concentration of cadmium ion is 10 -5 M.

[0032] (2) Add the precursor solution obtained in step (1) to an ultrasonic atomizer. After ultrasonic atomization, the precursor solution exists in the form of submicron droplets to obtain the precursor solution of water-soluble quantum dots. mist.

[0033] (3) Feed in nitrogen, the flow rate of nitrogen gas is 5L / min, and the droplets of the precursor solution obtained in step (2) are brought into the temperature-stabilized reaction device, and the droplets of the precursor solution are heated (the temp...

Embodiment 2

[0040] 1. The preparation of cadmium telluride quantum dots is the same as in Example 1.

[0041] 2. Detection of lactate dehydrogenase activity by cadmium telluride quantum dot fluorescence:

[0042] (1) Prepare the quantum dot solution: disperse the cadmium telluride quantum dots in 0.01 mol / L phosphate buffer (pH value is 6.8), the molar concentration of the cadmium telluride quantum dots is 8.9×10 -6 mol / L.

[0043] (2) preparation reaction substrate solution: n-butyl lactate is dissolved in the phosphate buffered saline (pH value is 6.8) of 0.01 mol / liter, and the molar concentration of n-butyl lactate is 5.2 * 10 -2 mol / L.

[0044] (3) Prepare the coenzyme solution: dissolve the oxidation state of nicotinamide adenine dinucleotide in 0.01 mol / L phosphate buffer (pH 6.8), the molar concentration of the oxidation state of nicotinamide adenine dinucleotide 3.6×10 -2 mol / L.

[0045] (4) Mix the solution prepared by step (1), step (2) and step (3), add deionized water to...

Embodiment 3

[0048] 1. The preparation of cadmium telluride quantum dots is the same as in Example 1.

[0049] 2. Detection of lactate dehydrogenase activity by cadmium telluride quantum dot fluorescence:

[0050] (1) Prepare the quantum dot solution: disperse the cadmium telluride quantum dots in 0.01 mol / L phosphate buffer (pH value is 6.8), the molar concentration of the cadmium telluride quantum dots is 2.6×10 -7 mol / L.

[0051] (2) Preparation of reaction substrate solution: n-butyl lactate is dissolved in 0.01 mol / liter of phosphate buffer (pH value is 6.8), and the molar concentration of n-butyl lactate is 6.2×10 -3 mol / L.

[0052] (3) Prepare the coenzyme solution: dissolve the oxidation state of nicotinamide adenine dinucleotide in 0.01 mol / L phosphate buffer (pH 6.8), the molar concentration of the oxidation state of nicotinamide adenine dinucleotide 5.8×10 -3 mol / L.

[0053] (4) Mix the solution prepared by step (1), step (2) and step (3), add deionized water to dilute, whe...

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Abstract

The invention belongs to the field of biological detection, in particular relates to a method for detecting enzymatic activity by using quantum dot fluorescence. The method provided by the invention is used for detecting the enzymatic activity through measuring the influence of a reaction system to water-soluble quantum dot fluorescence property in the enzyme reaction process. The method concretely comprises the following steps of: adding a certain quantity of enzymes to be detected in a reaction system containing water-soluble quantum dots or further in a reaction system containing coenzymes, enabling the fluorescence intensity of the water-soluble quantum dots to be varied due to the reaction of the enzymes to be detected, and realizing the high selective quantitative detection on the activity of the enzymes to be detected through the variable quantity of the fluorescence intensity of the water-soluble quantum dots. The method provided by the invention is simple in operation, rapid to detect and low in cost and can be used for detecting the activity of various physiological enzymes relevant to disease diagnosis.

Description

technical field [0001] The invention belongs to the field of biological detection, in particular to a method for quantum dot fluorescence detection of enzyme activity. Background technique [0002] Early and rapid detection of diseases is the key to the treatment of many major diseases. The earlier the correct diagnosis is made, the more conducive to the cure of the disease. Therefore, the development of early detection technology has received widespread attention from the medical community, and the familyization of disease detection and monitoring equipment has become the development of medicine. One of the important trends. In the latest early diagnosis methods, the detection of disease markers is a research hotspot. Taking cancer as an example, since the late 1970s, when Leon and Shapiro first confirmed that there is a high level of DNA in the peripheral blood circulation of tumor patients, and the level of tumor metastasis has a further increase trend, the discovery and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 唐芳琼任湘菱杨柳青
Owner TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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