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Method for preparing quantum dot labeled anti-porcine virus antibody immunochromatographic test strip, prepared test strip and application

An immunochromatographic test paper and quantum dot technology, applied in the field of immunoassays, can solve the problems of undeveloped veterinary swine infectious diseases, increase the difficulty of detection, time-consuming and labor-intensive problems, achieve simple detection procedures, improve sensitivity, and avoid abnormal specific effect

Pending Publication Date: 2020-09-15
北京中科基因技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The commercialized quantum dot immunochromatographic technology products are currently mainly human medicine products such as procalcitonin (PCT) quantitative assay kits produced by Shenzhen Jinzhun Biomedical Engineering Co., Ltd. For human serum, plasma, whole blood or peripheral blood, the detection can be completed even without the need for sample treatment solution to lyse the target in the sample to be tested, and the quantum dot immune chromatography technology products related to veterinary porcine infectious diseases have not been developed yet
[0004] However, when the quantum dot immunochromatography technology is applied to the detection of veterinary disease antigens, since there are many impurities or animal metabolites in the target of veterinary disease detection in the clinical test samples, it is relatively complicated, and most of them are metabolites or excreta such as Nasal swabs, anal swabs, feces, intestinal contents, and tissue samples, etc., are greatly affected by the sample treatment solution, which further increases the difficulty of detection
[0005] In the prior art, for the detection of various viral antigens that infect pigs, it is often necessary to prepare kits containing specific antibodies for detection. At the same time, the prepared kits also need to explore various conditions and Therefore, it is of great practical significance to provide a preparation method for immunochromatographic test strips that can adapt to a variety of specific antibody pairs for a variety of virus antigens that infect pigs

Method used

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  • Method for preparing quantum dot labeled anti-porcine virus antibody immunochromatographic test strip, prepared test strip and application
  • Method for preparing quantum dot labeled anti-porcine virus antibody immunochromatographic test strip, prepared test strip and application
  • Method for preparing quantum dot labeled anti-porcine virus antibody immunochromatographic test strip, prepared test strip and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1 Optimization of Quantum Dot Labeled Antibodies and Related Solutions

[0076] 1.1 Selection of maximum absorption peak of quantum dots

[0077] Purchase a number of commercial quantum dots, the maximum absorption peak is between 500-700nm, systematically label and explore the antibodies of different animal species, the following only the maximum absorption peaks are 500nm, 520nm, 525nm, 540nm, 565nm , 585nm, 605nm, 625nm, 655nm, 660nm, and 700nm quantum dots are used as examples for illustration.

[0078] Quantum dots with different maximum absorption peaks were mixed with porcine epidemic diarrhea virus monoclonal antibody PEDV-McAB2 at a molar ratio of 1:10 and then labeled. The labeling process was as follows: ①The activator EDC and quantum dots were mixed at a molar ratio of 2000:1 Mix the PBS buffer at pH 7.4 and shake it at room temperature for 1 hour to activate the quantum dots; ②mix the activated quantum dots with the antibody to be labeled at a mola...

Embodiment 2

[0110] The preparation of embodiment 2 kit

[0111] 2.1 Preparation of test strips

[0112] 2.1.1 Preparation of quantum dot marker pads

[0113] The reconstituted quantum dot-labeled antibody prepared in Example 1 is identified and then adsorbed to a glass fiber membrane or a polyester film. After drying, it becomes a quantum dot-labeled mat that can be used.

[0114] In order to evaluate the influence of the concentration of the quantum dot-labeled antibody in the preparation of the quantum dot-labeled pad, the quantum dot-labeled antibody was diluted with the quantum dot-labeled antibody reconstitution solution on the basis of A4, B4, C7, and D6 to a working concentration of 2.5, 5, After 8, 10, 20, 50, 100, 140, 150, 200 μg / ml, test strips were prepared and tested. The results (see Table 7): when the working concentration of the quantum dot-labeled antibody is 2.5 μg / ml, 5 μg / ml , 150 μg / ml, and 200 μg / ml, the sensitivity of the test strips to detect PEDV was low (10 4....

Embodiment 3

[0168] Embodiment 3 Preparation and application of other pig quantum dot test strips

[0169] In order to further evaluate whether each condition optimized in Example 2 can meet the application in the diagnosis and detection of animal diseases such as other pig antigens, porcine transmissible gastroenteritis virus, porcine circovirus type 2, porcine pseudorabies virus, etc. After debugging and evaluation, the results: (1) The sensitivity can be significantly improved by about 50 times or even higher than that of commercial colloidal gold detection strips, which solves the problem of no rapid detection products for porcine circovirus type 2 and porcine pseudorabies virus; (2) the specificity is good, and no non-specific phenomenon occurs; (3) the repeatability is good; (4) the storage period is much higher than the storage period (18 months) of commercial human quantum dot test strips; (5) ) has a good clinical application effect, and can be diagnosed quickly, on the spot and a...

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Abstract

The invention provides a method for preparing a quantum dot labeled anti-porcine virus antibody immunochromatographic test strip. According to the method, by optimization of the ratio of the quantum dots to an anti-porcine virus antibody, an antibody confining liquid, an antibody reconstitution fluid, a labeled antibody and a fixed antibody, high-sensitivity detection of the virus antigen in a sample is realized, and the sensitivity of the method is 50 times higher than that of a colloidal gold immunochromatography test strip. The preparation method disclosed by the invention is universal forvarious anti-porcine virus antibody pairs, and the quantum dot labeled immunochromatographic test strip prepared from various anti-porcine virus antibody pairs can sensitively detect virus antigens; meanwhile, the storage life can be prolonged to 27 months. The invention also relates to the quantum dot labeled immunochromatographic test strip prepared by the method, and a kit containing the quantum dot labeled immunochromatographic test strip.

Description

technical field [0001] The invention relates to the field of immunoassays, in particular to a method for preparing an anti-porcine virus antibody immunochromatographic test strip containing sub-dot markers, the prepared test strip, a kit and an application. Background technique [0002] Quantum dots (Quantum dot, QD), also known as semiconductor nanocrystals, are nano-fluorescent materials composed of II-VI or III-V elements with a radius smaller than or close to the exciton Bohr radius, mostly spherical. As a fluorescent material, quantum dots have excellent fluorescence characteristics: they have a narrow and symmetrical fluorescence emission spectrum, the emission wavelength varies with the material and particle size, they have a broad and continuous absorption spectrum, and the Stokes shift is large, so that Quantum dots are easy to realize the multicolor labeling function of single-wavelength excitation and simultaneous emission of multiple wavelengths. In addition, co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/558
CPCG01N33/6854G01N33/577G01N33/558G01N2333/47
Inventor 田克恭王莹晋春霞张许科
Owner 北京中科基因技术股份有限公司
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