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Method for preparing up-conversion aptamer test strip for rapid detection of ochratoxin A

A technology of ochratoxin and aptamer, which is applied in the field of preparation of up-conversion aptamer test strips, can solve problems such as complex process, long antibody preparation cycle, time-consuming and laborious preparation of test strips, etc., to reduce production costs, Effect of reducing background noise and improving sensitivity

Active Publication Date: 2018-11-30
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the preparation period of the antibody itself is long and the process is complicated, so the preparation of the test strip is time-consuming and laborious, and only qualitative detection can be realized.
Chinese patent 201710641207.0, colloidal gold chromatography rapid diagnostic test strip for detecting acetylcholinesterase and its preparation method, this technology uses colloidal gold as a marker and antibody as a recognition object, which can realize the rapid detection of acetylcholinesterase, but Unable to achieve quantitative goals
However, the recognition substance used is still an antibody, which increases the difficulty and cost of its preparation

Method used

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  • Method for preparing up-conversion aptamer test strip for rapid detection of ochratoxin A
  • Method for preparing up-conversion aptamer test strip for rapid detection of ochratoxin A
  • Method for preparing up-conversion aptamer test strip for rapid detection of ochratoxin A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Synthesis of Upconversion Luminescent Nanomaterials UCNPs 1 : Weigh YbCl 3 ·6H 2 O, YCl 3 ·6H 2 O and ErCl 3 (78%Y 3+ , 20%Yb 3+ , 2% Er 3+ ) was added to a 100mL three-necked flask, and 6mL oleic acid and 15mL octadecene were added. Under stirring and nitrogen atmosphere, gradually raise the temperature to 160°C to make the reagents form a uniform solution, and cool naturally to room temperature. Weigh 4mmol NH 4 F and 2.5mmol NaOH were dissolved in 10mL methanol solution. The above solution was added dropwise into a three-necked flask, heated up to 60° C. and magnetically stirred for 30 minutes, and then gradually heated up to remove methanol by evaporation. Continue to flow argon, add a condensing reflux device, raise the temperature to 300°C, and continue the reaction for 1h. After the reaction was over, the device was closed and cooled to room temperature naturally. The material was centrifuged and washed three times with water and ethanol, and dried at...

Embodiment 2

[0041] Synthesis of Upconversion Luminescent Nanomaterials UCNPs 2 , the specific method is as follows: add 0.3g NaOH, 1.5mL deionized water, 5mL oleic acid and 10mL ethanol into a 100mL beaker in sequence, stir and mix quickly to form a transparent and uniform solution. Weigh 1.6mL Y(NO 3 ) 3 ·6H 2 O(0.5mol / L), 0.9mL Yb(NO 3 ) 3 ·5H 2 O (0.2mol / L), 0.1mL Yb (NO 3 ) 3 ·5H 2 O (0.2mol / L) was added dropwise to the above solution, and stirred vigorously to mix. Weigh 0.168g NaF, dissolve it in 4mL deionized water, and add it dropwise to the above mixed solution. Stir vigorously for 15 min, then transfer the mixture to a 100 mL polytetrafluoroethylene-lined reactor, and react at 200 °C for 8 h. After the reaction, the reactor was taken out to cool down to room temperature naturally, and the bottom precipitate was collected, washed three times with ethanol centrifugation, and dried at 70°C for 8 hours to obtain a white solid powder, which was stored for future use.

[00...

Embodiment 3

[0044] Synthesis of Upconversion Luminescent Nanomaterials UCNPs 3 , the specific method is as follows: take 0.18g Y 2 o 3 (28%), 0.788gYb 2 o 3 (70%) and 0.022g Er 2 o 3 (2%), respectively added into the nitric acid solution, heated to dissolve, and evaporate excess nitric acid to obtain the nitrate powder of the above rare earth element. After dissolving in 8 mL of deionized water, add 2.1273 g of ethylenediaminetetraacetic acid (EDTA) to adjust the pH of the solution to be weakly alkaline and fully dissolve. Measure 25mL of ethylene glycol solution, add 0.4g of cetyltrimethylammonium bromide (CTAB) and the above mixture, and stir rapidly. 1.5 mL HF was added dropwise. After the solution turns into a white milky colloid, add 3.5mL of concentrated nitric acid, stir evenly, then transfer to a 100mL reaction kettle with a polytetrafluoroethylene liner, and react at 195°C for 24h. After the reaction was finished, the device was closed, and the reactor was taken out, and ...

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Abstract

The invention discloses a method for preparing an up-conversion aptamer test strip for rapid detection of ochratoxin A. The method comprises the following steps: modifying an up-conversion luminescentnano material, to be more specific, adding oleic acid and octadecene to an inorganic salt containing Yb<3+>, Y<3+> and Er<3+>, stirring, introducing an inert gas, and heating to form a uniform solution; dissolving ammonium fluoride and sodium hydroxide in methanol, adding the ammonium fluoride and sodium hydroxide methanol solution in the uniform solution, removing the methanol by heating and evaporation, introducing an inert gas to carry out reaction, adding a reaction product into a condensing reflux device, and heating to carry out reaction; mixing polyacrylic acid and ethanol to obtain apolyacrylic acid and ethanol mixed solution, dispersing the up-conversion luminescent nano material in a chloroform solution, adding the up-conversion luminescent nano material into the polyacrylic acid and ethanol mixed solution, and stirring for reaction to obtain a modified up-conversion luminescent nanomaterial; and preparing an up-conversion aptamer probe. The invention provides a new chromatographic test strip technology which can effectively improve the sensitivity of the test strip, reduce the production cost, and achieve the purpose of quantitative detection.

Description

technical field [0001] The invention belongs to the technical field of mycotoxin detection, and in particular relates to a preparation method of an up-conversion aptamer test strip for rapid detection of ochratoxin A. Background technique [0002] Ochratoxin is another mycotoxin that has attracted worldwide attention after aflatoxin. It is a group of important food-contaminating mycotoxins produced by 7 species of Aspergillus of the genus Aspergillus and 6 species of Penicillium of the genus Penicillium, of which 4 are the most toxic, the most widely distributed, the highest toxin production, and the most harmful to agricultural products. Ochratoxin A is the most polluted and most closely related to human health. Chinese patent 201611111887.7, a potato virus rapid detection test strip and its preparation method and application, the test strip uses antibodies as recognition substances to achieve the purpose of target detection. However, the preparation cycle of the antibody...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/558
CPCG01N33/54306G01N33/54346G01N33/558
Inventor 吴世嘉刘丽红段诺王周平虞倩茹
Owner JIANGNAN UNIV
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