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Portable high-sensitivity ractopamine immunoassay determination kit and application thereof

A ractopamine and ractopamine nanotechnology, applied in the field of food safety detection immunoassay, can solve problems such as inability to apply ractopamine, and achieve the effects of ensuring sensitivity, good reproducibility, and ease of operation and production

Pending Publication Date: 2022-01-14
INST FOR AGRI APPL OF ATOMIC ENERGY SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The kit is detected through a competitive reaction system, which overcomes the defect that the "sandwich" structure model adopted by the current immunoassay method is basically not suitable for the detection of ractopamine

Method used

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  • Portable high-sensitivity ractopamine immunoassay determination kit and application thereof
  • Portable high-sensitivity ractopamine immunoassay determination kit and application thereof
  • Portable high-sensitivity ractopamine immunoassay determination kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Preparation method of ractopamine portable immunoassay assay kit

[0055] S13. Preparation of ractopamine nano-gold probes

[0056] (1) Synthesis method of gold nanoparticles

[0057] Add 98mL of deionized water to the siliconized beaker respectively, and then add 2mL of 50mM chloroauric acid stock solution respectively, so that the concentration of chloroauric acid is 1mM, stir on a magnetic stirrer at 1000rpm / min, heat at 250°C, continue to boil after the liquid boils Boil for 2 minutes; absorb 10mL of 38.8mM trisodium citrate, quickly add it to the beaker at one time, keep the stirring speed and heating temperature constant, and continue to boil for 6 minutes. It can be found that the color of the solution gradually changes from colorless to purple, and then into wine red. Depending on the amount of trisodium citrate added, it finally turns into a different color. After the liquid is cooled, the original volume is restored with deionized water, and it is stored in a...

Embodiment 2

[0084] Kit use method of the present invention

[0085] The specific operation of the ractopamine nano-gold composite probe prepared in the above implementation case 1 combined with the portable blood glucose meter immunoassay assay kit is as follows:

[0086] S19. Take out the kit from the refrigerator at 4°C, and equilibrate at room temperature for 10 minutes;

[0087] S20. Add ractopamine standards (10.0 ng L −1 - 40 µg L −1 ) and 20 μL of the sample obtained after pretreatment, set up a duplicate, then add 20 μL of magnetic nanoprobes to each well, shake and mix well with a micro shaker, and then place it at a temperature of 37°C and a relative humidity of 70%. Incubate for 1 h in a constant temperature and humidity incubator;

[0088] S23. Wash three times with PBST solution, add 0.5 μL (units / μL) Phi29 DNA polymerase and 0.5 μL (10 mmol / L) dNTP mixture;

[0089] S24, add 1 μL ssDNA-invertase polymer and incubate for 1 hour, add 200 μL (0.5mol / L) sucrose solution aft...

experiment example 3

[0093] The methodological examination result of kit of the present invention

[0094] The kit prepared in Example 1 was identified according to the common verification procedures in this field, and the results are shown in Table 1.

[0095] Table 1 The methodological examination result of kit of the present invention

[0096]

[0097] The above results show that the accuracy, specificity, precision, sensitivity and stability of the "Ractopamine Nano Gold Probe Combined with Portable Blood Glucose Meter Immunoassay Assay Kit" are fully qualified.

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Abstract

The invention relates to the technical field of food safety detection immunoassay, in particular to a portable high-sensitivity ractopamine immunoassay determination kit and application thereof. The kit comprises a ractopamine standard substance, a ractopamine magnetic nanoprobe, a ractopamine nanogold nanoprobe and an ssDNA-invertase conjugate. The kit can be used for specifically and quantitatively detecting the content of ractopamine. The kit has the advantages of simplicity, rapidness, sensitivity, specificity, stability and the like. Moreover, the detection system provided by the invention is operated in an open manner, is simple, convenient and rapid, is particularly suitable for being popularized and used by vast quality inspection institutions, and provides a very valuable detection means for food safety detection. The kit provided by the invention is combined with a portable glucometer for use, the sensitivity is greatly improved by detecting a glucose signal value corresponding to the rolling circle DNA, and a more sensitive, rapid and reliable basis can be provided for detection of ractopamine in food.

Description

technical field [0001] The invention relates to the technical field of immunoassay for food safety detection, in particular to a portable high-sensitivity immunoassay assay kit for ractopamine and its application. Background technique [0002] Pesticides and veterinary drugs are indispensable and important means of production for agricultural production, and play an important role in ensuring agricultural and food production and safety. In the foreseeable future, chemical pesticides and veterinary drugs will still be an indispensable means of ensuring the quality and safety of agricultural products. Residues of pesticides and veterinary drugs in food and their safety assessment are widely concerned by scientific researchers and the public. However, if it is used unreasonably for a long time, it will cause environmental pollution, ecological damage and harm to human health. Therefore, the establishment of efficient, rapid and low-cost veterinary drug residue detection has b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/577G01N33/66C12Q1/682C12Q1/6804
CPCG01N33/54326G01N33/54346G01N33/577G01N33/66G01N33/94C12Q1/6804C12Q1/682C12Q2531/125C12Q2563/155C12Q2521/50
Inventor 杜鹏飞贺锋王维婷柳尧波王守经李腾飞马艳丽胡鹏
Owner INST FOR AGRI APPL OF ATOMIC ENERGY SHANDONG ACADEMY OF AGRI SCI
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