Fluorescence sensor as well as preparation method and application thereof

A fluorescent sensor and hydrogen bonding technology, applied in the field of biosensors, can solve the problems of complex pretreatment, long detection cycle, poor reproducibility, etc., and achieve the effects of good water solubility, simple detection method, and low toxicity

CN107014791AActive Publication Date: 2017-08-04ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2017-08-04

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Abstract

The invention belongs to the technical field of a biosensor and specifically relates to a fluorescence sensor as well as a preparation method and application thereof. A carbon quantum dot is utilized as a fluorescent material, a monoclonal antibody and an aptamer are simultaneously combined with a target object and the fluorescence sensor with a sandwich type structure is lastly acquired; the aptamer of the target object is arranged on the upper part of the fluorescence sensor; the target object is arranged on the middle part; the monoclonal antibody of the target object is arranged on the lower part; a sulfydryl group is modified on the aptamer of the target object; the carbon quantum dot is combined with the aptamer of the target object under the effect of hydrogen bonds. The invention also discloses a preparation method of the fluorescence sensor. The fluorescence sensor prepared according to the invention can be used for quantitatively determining the concentration of toxic fungus and toxin, especially quantitative determining the concentration of aspergillus flavus B1, has high stability and specificity, is wide in detection scope and is more accurate.
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Description

technical field

[0001] The invention belongs to the technical field of biosensors, and in particular relates to a fluorescent sensor and its preparation method and application. Background technique

[0002] Aflatoxins (AFT) are highly toxic substances. In 1993, it was classified as a Class 1 carcinogen by the Cancer Research Institute of the World Health Organization (WHO). The danger of aflatoxin is that it can damage the liver tissue of humans and animals, and can lead to liver cancer or even death in severe cases. Aflatoxin B1 (AFB1 for short) is the most common in naturally contaminated food, and its toxicity and carcinogenicity are also the strongest. It can often be detected in corn, peanuts, cotton seeds and some dried fruits. At present, the development of highly sensitive aflatoxin B1 detection method has become the focus of international attention.

[0003] Nucleic acid aptamers can bind to specific target substances, have the characteristics of high specificity...

Examples

Embodiment 1

[0040] A fluorescent sensor, the fluorescent sensor is a sandwich structure, with carbon quantum dots as the fluorescent material; the upper part of the sandwich structure is the nucleic acid aptamer of the target, the middle part of the sandwich structure is the target, and the sandwich structure The lower part is the monoclonal antibody of the target object, the nucleic acid aptamer of the target object is modified with sulfhydryl groups, and the carbon quantum dots are bound to the nucleic acid aptamer of the target object through hydrogen bonding.

Embodiment 2

[0042] Taking the target object as aflatoxin B1 as an example, the target object of the present invention is not limited to aflatoxin B1.

[0043] A preparation method of a fluorescent sensor comprises the following steps:

[0044] Step 1: Preparation of carbon quantum dot-nucleic acid aptamer detection probe

[0045] 1) According to the volume ratio of glucose aqueous solution and NaOH solution as 1:1, mix 10mL, 1mol / L glucose aqueous solution with 10mL, 1.5mol / L NaOH solution, and sonicate for 4h under 400w ultrasonic power; adjust the pH of the mixture to 7. Add 100ml of absolute ethanol dropwise and stir, then add magnesium sulfate with a mass fraction of 12%, and stir for 20 minutes to obtain a carbon quantum dot solution, store it for 24 hours, and set aside. The particle size of the carbon quantum dots in the carbon quantum dot solution is 4~5nm.

[0046] Carbon quantum dots are an environmentally friendly fluorescent material. Compared with other fluorescent material...

Embodiment 3

[0057] An application of a fluorescent sensor in the quantitative detection of toxins, taking the detection of aflatoxin B1 as an example, but the present invention is not limited to the detection of aflatoxin B1, and the detection method includes the following steps:

[0058] Step 1: Prepare aflatoxin B1 test solution

[0059] 1) Preparation of aflatoxin B1 mother solution

[0060] First, dissolve 1 mg of aflatoxin B1 standard substance in 1 mL of methanol-PBS (where the mass fraction of methanol is 10%) solution to obtain solution A, and the concentration of solution A is C A =1mg / mL;

[0061] Take 1 μl of solution A and add methanol-PBS (the mass fraction of methanol is 10%) solution to dilute to 100mL of solution B, the concentration of solution B is C B =10ng / mL, 100mL, 10ng / mL of aflatoxin B1 mother solution.

[0062] 2) Dilute the aflatoxin B1 mother solution to obtain different concentrations of aflatoxin B1 test solution

[0063] Take 10 μL of aflatoxin B1 mother ...