A time-resolved fluorescent immunochromatographic test strip for detecting acetamiprid and its preparation method and application

A time-resolved fluorescence and immunochromatographic test paper technology, applied in analytical materials, biological testing, fluorescence/phosphorescence, etc., can solve the problems of high professional requirements for operators, expensive detection costs, cumbersome steps, etc., and increase the sensitivity of the reaction. , The effect of short detection time and simple operation

Active Publication Date: 2021-09-03
ZHENGZHOU TOBACCO RES INST OF CNTC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the above methods require advanced detection instruments, the detection costs are expensive, the steps are cumbersome, time-consuming, and require high professionalism of operators, they are not suitable for high-throughput rapid screening detection in grassroots enterprises and institutions.

Method used

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  • A time-resolved fluorescent immunochromatographic test strip for detecting acetamiprid and its preparation method and application
  • A time-resolved fluorescent immunochromatographic test strip for detecting acetamiprid and its preparation method and application
  • A time-resolved fluorescent immunochromatographic test strip for detecting acetamiprid and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Composition of time-resolved fluorescent immunochromatographic test strips for detecting acetamiprid

[0041] 1. Test strips

[0042] see figure 1 : The test strip is composed of a bottom plate, a sample absorbent pad, a conjugate release pad, a nitrocellulose membrane and an absorbent pad;

[0043] The sample absorbent pad 1, the conjugate release pad 2, the nitrocellulose membrane 3 and the water absorbent pad 4 are overlapped and pasted on the bottom plate 7 in sequence, and 1 / 3 of the conjugate release pad is covered by the sample absorbent pad from the starting end , the end of the conjugate release pad is connected to the beginning of the nitrocellulose membrane, the end of the nitrocellulose membrane is connected to the beginning of the absorbent pad, the beginning of the sample absorbent pad is aligned with the beginning of the PVC bottom plate, and the end of the absorbent pad is connected to the end of the PVC bottom plate alignment;

[0044] The ni...

Embodiment 2

[0046] Example 2 Preparation of time-resolved fluorescent immunochromatographic test strips for detecting acetamiprid

[0047] The preparation method of the time-resolved fluorescent immunochromatographic test strip for detecting acetamiprid mainly includes the following steps:

[0048] 1) Preparation of the conjugate release pad: label the acetamiprid monoclonal antibody with commercially available fluorescent microspheres, and dilute it with a specific buffer system, soak the conjugate release pad in the dilution buffer, and vacuum freeze-dry Post-preparation;

[0049] 2) Preparation of nitrocellulose membrane: Spray the acetamiprid hapten-carrier protein conjugate on the detection area of ​​the nitrocellulose membrane to make the detection area; spray goat anti-mouse anti-antibody onto the nitrocellulose membrane The scope of the quality control area above is made into the quality control area;

[0050] 3) Assembly and cutting: Paste the sample absorption pad, the conjuga...

Embodiment 3

[0103] Example 3 Application of time-resolved fluorescent immunochromatographic test strips for detecting acetamiprid

[0104] 1. Tobacco sample pretreatment

[0105] Weigh 1.0±0.05 g of crushed tobacco leaf sample into a polystyrene centrifuge tube; add 5 mL of 50% methanol aqueous solution, and use a homogenizer to fully break it up to obtain a sample liquid; pipette 75 μL of sample liquid and 425 μL of The ionized water was mixed and waited for inspection.

[0106] 2. Test with test strips

[0107] Draw 100 μL of the sample solution to be tested and drop it vertically into the sample hole of the test paper card. When the liquid flows, start timing and react for 10 minutes; insert the test paper card into the carrier of the KFT-100A fluorescence detector, and select the sample to be Check the items, press the "Start Test" button, the fluorescence detector will automatically scan the test paper card, and read or print the test results on the display screen of the instrument...

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Abstract

A time-resolved fluorescent immunochromatographic test strip for detecting acetamiprid, its preparation method and application. The test strip includes a bottom plate and a sample absorbent pad, a conjugate release pad, a nitrocellulose membrane, and a water absorbent pad that are sequentially overlapped and pasted on the bottom plate, and the conjugate release pad is embedded with acetamiprid monoclonal antibody labeled with fluorescent microspheres. , the detection area and the quality control area are fixed on the nitrocellulose membrane, the detection area is sprayed with acetamiprid hapten-carrier protein conjugate, the quality control area is sprayed with goat anti-mouse anti-antibody, and the acetamiprid hapten is made of E - N ‑(4‑(6‑chloropyridine‑3‑yl)2‑butene‑2‑yl)cyanamide reacts with aminopropionic acid. The invention also provides a preparation method of the test strip and a method for detecting acetamiprid in a sample by using the test strip. The test strip and detection method provided by the invention have the advantages of simple operation, high sensitivity, fast detection speed and low cost, and can realize rapid detection and on-site monitoring of acetamiprid in large batches of samples.

Description

technical field [0001] The invention belongs to the field of pesticide residue detection, and in particular relates to a time-resolved fluorescent immunochromatographic test strip for detecting acetamiprid in tobacco and tobacco products, a preparation method and application thereof. Background technique [0002] Acetamiprid belongs to neonicotinoid insecticides, which are an important class of insecticides after organophosphorus, carbamate and pyrethroid insecticides. By selectively controlling the nicotinic acetylcholinesterase receptors (nAChRs) in the central nervous system of insects, the normal conduction of the central nervous system of insects is blocked, resulting in general paralysis and death of the pests. Acetamiprid is a representative agent of neonicotinoid insecticides. It is widely used in agricultural production to prevent and control various pests in seeds, leaves and soil. The effect of pest control is ideal, and it is mainly used for the control of tobac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558G01N33/58G01N33/533G01N21/64
CPCG01N21/6408G01N33/533G01N33/558G01N33/582G01N33/585
Inventor 陈黎范子彦刘惠民唐纲岭崔华鹏樊美娟赵乐郭吉兆聂聪潘立宁
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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