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Quantum dot immunochromatographic test strip for rapidly detecting pyrimethanil residues and preparation method

An immunochromatographic test paper and quantum dot technology, which is applied in the field of food safety immunology detection, can solve the problems that pyrimethanil has not been reported, and achieve the effects of shortening the detection time, high sensitivity, and protecting stability

Inactive Publication Date: 2020-11-03
烟台市疾病预防控制中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, quantum dots have been widely used as fluorescent markers to detect proteins and other small molecules, and are also widely used in the detection of pesticide residues, but there is no report on the detection of pyrimethanil residues

Method used

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  • Quantum dot immunochromatographic test strip for rapidly detecting pyrimethanil residues and preparation method
  • Quantum dot immunochromatographic test strip for rapidly detecting pyrimethanil residues and preparation method
  • Quantum dot immunochromatographic test strip for rapidly detecting pyrimethanil residues and preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1: A quantum dot immunochromatographic test strip for rapid detection of pyrimethanil residues, the structure of the test strip in this embodiment is as follows figure 1 As shown, it includes the bottom plate, and the sample pad 6, the conjugate release pad 5, the chromatographic membrane 2, and the water-absorbing pad 1 that are sequentially spliced ​​on the bottom plate; the conjugate release pad 5 is coated with pyrimethanil labeled with quantum dots antibody complex. The chromatographic membrane 2 is coated with a detection line 4 and a quality control line 3 parallel to the end surface of the bottom plate, the detection line is close to the sample pad, and the quality control line is close to the water-absorbing pad.

[0026] The quantum dots are quantum dot fluorescent microspheres, and the quantum dot fluorescent microspheres are core-shell nanocomposite particles wrapped with NHS-activated CdTe / ZnSe as the core and silica as the shell; and the surface of...

Embodiment 2

[0031] Embodiment 2: Preparation of immunochromatographic test strips

[0032] (1) Preparation of the sample pad. In this embodiment, the sample pad glass fiber membrane pretreatment solution is 1% BSA, 1% sucrose, 0.1% PEG, PBS buffer solution of 0.05% Tween-20, and the glass fiber membrane is soaked After 1 hour, dry it for later use.

[0033] (2) Prepare the conjugate release pad, that is, the quantum dot-labeled pyrimethanil monoclonal antibody fluorescent probe,

[0034] a. Centrifuge the water-soluble CdSe / ZnS core-shell quantum dots modified by surface carboxyl group at 12000rpm for 5min, b redissolve with 20mM Tris buffer, add activation reagent EDC ((1-(3-dimethylaminopropyl)-3 -ethylcarbodiimide) to 1.4mg / mL, N-hydroxysuccinimide to 2mg / mL. c, adding the anti-pyrimethanil monoclonal antibody with a final concentration of 40mg / L, stirring at room temperature for 3h, after the reaction , centrifuged at 12000rpm for 5min to obtain a precipitate, which was properly dil...

Embodiment 3

[0044] Embodiment 3: sample test and result judgment

[0045] (1) Sample processing

[0046] Choose one of the vegetable or fruit samples cabbage, cucumber, and tomato, and refer to (Yi Cao, Haixing Shi, Tao Le, Ran Tang & Yong Xie (2019) Development a monoclonal antibody-based enzyme-linked immunosorbent assay for screening pyrimethanil in fruits and vegetables, Food and Agricultural Immunology, 30:1, 548-563, DOI: 10.1080 / 09540105.2019.1608160). First, chop and homogenize the sample, weigh 5 g of the homogenized sample into a 50 mL polystyrene centrifuge tube, add 5 mL of acetonitrile, and shake evenly. Then add 2g NaCl, 4g anhydrous magnesium sulfate. The mixture was centrifuged at 10,000 rpm for 5 min, the supernatant was concentrated with a nitrogen blower, and finally dissolved to 1 mL with PBS buffer. For each test, take 50 μL of the sample extract and add it dropwise to the sample pad.

[0047] figure 2 It shows the result judgment chart of the test results of th...

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Abstract

The invention provides a quantum dot immunochromatographic test strip for rapidly detecting pyrimethanil residues and a preparation method. The immunochromatographic test strip comprises a bottom plate, and a sample pad, a conjugate release pad, a chromatographic membrane and a water absorption pad which are sequentially spliced on the bottom plate, wherein the conjugate release pad is coated witha pyrimethanil antibody compound marked by quantum dots. Compared with an ELISA detection method of similar antibodies, the ELISA detection method disclosed by the invention has the advantages that the detection time is greatly shortened, the detection efficiency is improved, the detection sensitivity is higher, and quantitative detection can be realized on pyrimethanil residues in a sample.

Description

technical field [0001] The invention relates to the technical field of food safety immunological detection, in particular to a pyrimethanil residual quantum dot immunochromatographic test strip and a preparation method thereof. Background technique [0002] Pyrimethanil belongs to the broad-spectrum fungicides of anilinopyrimidines. By inhibiting the biosynthesis of methionine, it affects cell division and protein formation, thereby preventing the infection of pathogens. It has root systemic and leaf penetration activities. . Due to its excellent broad-spectrum fungicidal properties, pyrimethanil is widely used in vegetables, fruits and ornamental plants to control post-harvest diseases such as Botrytis cinerea. However, due to the wide application of pyrimethanil in horticultural species, long-term studies have shown that it is potentially carcinogenic to aquatic organisms such as mice and rats. Canada determined the maximum residues of pyrimethanil in tomatoes and onions ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/577G01N33/558G01N33/53G01N33/531
CPCG01N33/5308G01N33/531G01N33/558G01N33/577G01N33/582G01N33/588
Inventor 张晓瑜张桂芳邹晓楠王志昱张艺王颖刘泽静张甜甜王乙惠董桂贤
Owner 烟台市疾病预防控制中心
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