Up-conversion fluorescent aptamer-based side stream test paper detection method

A detection method and aptamer technology, which is applied in the field of environmental detection and food safety detection, can solve the problems of high price, instability, and expensive antibody preparation, and achieve the effect of low equipment requirements, high sensitivity, and strong specificity

Active Publication Date: 2018-06-22
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its biggest limitation is: antibody preparation is expensive, expensive and unstable, powerless for toxic or non-immune targets
Relevant workers have conducted a lot of research based on aptamers, but a fast, sensitive and portable multi-target detection method based on aptamers has not yet appeared.

Method used

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  • Up-conversion fluorescent aptamer-based side stream test paper detection method
  • Up-conversion fluorescent aptamer-based side stream test paper detection method
  • Up-conversion fluorescent aptamer-based side stream test paper detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Below in Hg 2+ The detection of is example, illustrates the effect of the inventive method, concrete experiment process, comprises the following steps:

[0048] 1) YCl 3 ·6H 2 O (267.0mg, 0.88mmol), TmCl 3 ·6H 2 O (7.7mg, 0.02mmol), ErCl 3 Dissolve 6H2O (38.2 mg, 0.1 mmol) in 2 mL of deionized water, add 7.5 mL of oleic acid and 15 mL of octadecene, stir at room temperature for 30 minutes, then slowly heat to 120 ° C, keep the temperature for 1 hour and then heat to 156 ° C for 1 hours, dehydrated under the protection of argon and cooled to room temperature. Add dissolved NH 4 F (148.15 mg, 4 mmol) and NaOH (100 mg, 2.5 mmol) in methanol 10 mL were stirred at room temperature for 2 hours. After methanol volatilized, the solution was heated to 280°C for 1.5 hours and then cooled to room temperature. Wash with ethanol and cyclohexane for 3 to 5 times, store in cyclohexane to obtain β-NaYF with a particle size of 35nm 4 :Er / Tm;

[0049] 2) 14.5 μL of polypropylen...

Embodiment 2

[0064] Taking the detection of ochratoxin A as an example below, the effect of the inventive method is illustrated, and the specific experimental process includes the following steps:

[0065] 1) YCl 3 ·6H 2 O (242.69mg, 0.8mmol), ErCl 3 ·6H 2 O (7.64mg, 0.02mmol) and YbCl 3 Dissolve 6H2O (69.75mg, 0.18mmol) in 2mL deionized water, add 7.5mL oleic acid and 15mL octadecene, stir at room temperature for 30 minutes, then slowly heat to 120°C for 1 hour, then heat to 156°C for 1 hour , dehydrated under argon protection and cooled to room temperature. Add dissolved NH 4 F (148.15 mg, 4 mmol) and NaOH (100 mg, 2.5 mmol) in methanol 10 mL were stirred at room temperature for 2 hours. After methanol volatilized, the solution was heated to 280°C for 1.5 hours and then cooled to room temperature. Wash with ethanol and cyclohexane for 3-5 times, store in cyclohexane to obtain β-NaYF with a particle size of 45nm 4 :Yb / Er;

[0066] 2) 14.5 μL of polypropylene, 1 mL of ethanol and ...

Embodiment 3

[0081] Take the detection of Salmonella as an example below to illustrate the effect of the inventive method, and the concrete experimental process comprises the following steps:

[0082] 1) YCl 3 6H2O (210.8mg, 0.695mmol), YbCl 3 ·6H 2 O (116.2mg, 0.30mmol), and TmCl 3 ·6H 2 O (1.9mg, 0.005mmol). Dissolve in 2mL deionized water, add 7.5mL oleic acid and 15mL octadecene, stir at room temperature for 30 minutes, then slowly heat to 120°C for 1 hour, then heat to 156°C for 1 hour , dehydrated under argon protection and cooled to room temperature. Add dissolved NH 4 F (148.15 mg, 4 mmol) and NaOH (100 mg, 2.5 mmol) in methanol 10 mL were stirred at room temperature for 2 hours. After methanol volatilized, the solution was heated to 280°C for 1.5 hours and then cooled to room temperature. Wash with ethanol and cyclohexane for 3-5 times, store in cyclohexane to obtain β-NaYF with a particle size of 45nm 4 :Yb / Tm;

[0083] 2) 14.5 μL of polypropylene, 1 mL of ethanol and 1 ...

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Abstract

The invention discloses an up-conversion fluorescent aptamer-based side stream test paper detection method, and belongs to the technical fields of environment detection and food safety detection. A nucleic acid aptamer capable of specifically identifying target bacteria is modified on the surface of each nano-gold particle; cDNA which is in complementary pairing with the aptamer is modified on thesurface of each rare earth up-conversion fluorescent particle; and after the two kinds of nanoparticles are mixed, the basic groups of the two segments of nucleic acid sequences are in complementarypairing and the distance of the two kinds of particles are pulled closely, so that fluorescence of the upper-conversion fluorescent particles is quenched. After a target object is added, the target object can be bound with the aptamer with cDNA competitiveness, so that fluorescence recovery of the up-conversion nanoparticles is caused. According to different fluorescence recovery intensity, quantitative measurement of the target bacteria can be realized. The method provided by the invention is high in sensitivity, high in specificity and simple and convenient in operation, various target objects can be measured through change of the aptamer sequence, and very import significance in the aspects of environment monitoring, food analysis and the like is achieved.

Description

technical field [0001] The invention belongs to the technical field of environmental detection and food safety detection, and in particular relates to a lateral flow test paper detection method based on an up-conversion fluorescent aptamer. Background technique [0002] Rapid, sensitive, and portable detection methods have received more and more attention in the fields of disease diagnosis, environmental monitoring, and food safety testing. For example, toxic substances such as pathogens and heavy metal ions are important indicators of water quality evaluation; in addition, as one of the most concerned issues of global public health, foodborne diseases are often caused by different types of pollutants (such as bacteria, antibiotics, illegal additives and Pesticide residues) pollute food, which causes one tenth of the population to get sick every year; in addition, in disease detection, in addition, doctors usually need to evaluate the content of target substances such as bac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533
CPCG01N33/533
Inventor 林敏徐峰金碧瑞杨叶欣卢天健
Owner XI AN JIAOTONG UNIV
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