Thionine labeled antibody immunochromatography testing strip for visually and rapidly detecting transgenic protein

A technology of immunochromatographic test strips and labeled antibodies, applied in biological testing, measuring devices, analytical materials, etc., can solve the problems of complex synthesis and modification process, low sensitivity, high cost, and achieve good stability and reproducibility, The effect of high detection sensitivity and shortened time

Pending Publication Date: 2021-11-23
SHANGHAI ACAD OF AGRI SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The continuous update of markers improves the detection sensitivity of ICS, but most of these materials require complex synthesis and modification processes
[0005] Nanomaterials such as colloidal gold are widely used in the construction of test strips as color signals, but complex antibody labeling and material modification processes are required, and the sensitivity is generally low

Method used

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  • Thionine labeled antibody immunochromatography testing strip for visually and rapidly detecting transgenic protein
  • Thionine labeled antibody immunochromatography testing strip for visually and rapidly detecting transgenic protein
  • Thionine labeled antibody immunochromatography testing strip for visually and rapidly detecting transgenic protein

Examples

Experimental program
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Effect test

Embodiment 1

[0049] Example 1 Preparation method of thionine-labeled antibody immunochromatography test strip for visual and rapid detection of transgenic protein

[0050] 1. Thi optimum labeling concentration

[0051] Thionine Thi is used as the labeling signal in the experiment, and the labeling of the specific monoclonal antibody mAb1 directly affects the signal intensity of the detection line and the sensitivity of the test strip, and affects the concentration of Thi, labeling time and the final volume of labeled antibody required. Optimized.

[0052] At room temperature, the antibodies were labeled with Thi at a concentration of 0.1mg / mL, 0.25mg / mL, 0.5mg / mL, 1mg / mL, 1.5mg / mL and 2mg / mL respectively. The labeling time was 10min, and 5μL glutaraldehyde (5%) incubation, by observing the intensity of the signal in the detection area, select the Thi with clear and uniform bands and no background interference as the optimal labeling concentration.

[0053] Such as figure 1 As shown, wi...

Embodiment 2

[0080] In order to further verify the practicability and accuracy of the test strip of the present invention in actual samples, 6 kinds of soybean and 6 kinds of corn seed samples were randomly selected from the local market, and the samples were processed and protein extracted with the same method, and the protein concentration 10% of the supernatant was tested, the corresponding standard 10% soybean RRS (S), 5% corn NK603 (M) and the corresponding blank crop supernatant (N) were used as controls, and the experimental results were verified by the current standard PCR method. Transgenic soybean RRS and corn NK603 were used as positive controls, and blank soybeans and corn were used as negative controls.

[0081] Kits were used to extract the DNA of randomly purchased soybean and corn samples and set aside. The primer sequences of the EPSPS gene were selected: F1: 5'-GACTTGCGTGTTCGTTCTTC-3' and R1: 5'-AACACCGTTGAGCTTGAGAC-3'.

[0082] The corresponding system was added into th...

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Abstract

The invention relates to a thionine labeled antibody immunochromatography testing strip for visually and rapidly detecting transgenic protein, the thionine labeled antibody immunochromatography testing strip comprises a bottom plate, and a sample pad, a combination pad, a nitrocellulose membrane and absorbent paper which are fixed on the bottom plate and are connected in sequence, and a labeled antibody labeled by thionine is fixed in the combination pad; a Thi-mAb1 compound which is formed by coupling the thionine with a monoclonal antibody mAb1 through glutaraldehyde. The immunochromatography testing strip provided by the invention takes Thi as a signal marker, does not need a nano material, does not need a complex synthesis process, utilizes a conventional cross-linking agent to be rapidly coupled with an antibody through a covalent bond, breaks through the tedious preparation process of colloidal gold, greatly shortens the time for marking the antibody, simplifies the preparation process, and can rapidly detect transgenic protein; the sensitivity is high; and the specificity, the stability and the reproducibility are good.

Description

technical field [0001] The invention belongs to the technical field of rapid immunodetection, in particular to a thionine-labeled antibody immunochromatographic test strip for visual and rapid detection of transgenic proteins. Background technique [0002] At present, more and more detection techniques are applied to the detection of genetically modified crops. PCR is the main method to detect the specific DNA of transgenic crops, but its operation is cumbersome, time-consuming and requires expensive instruments, which is not suitable for on-site detection. LAMP and RPA overcome the drawbacks of PCR and can be detected quickly, sensitively, and with high specificity. However, they use agar gel electrophoresis, which is time-consuming and easy to contaminate, so there are still certain limitations. [0003] Although the above detection based on nucleic acid level has ultra-high sensitivity, there are still certain disadvantages in the actual sample on-site detection. The de...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/573G01N33/577G01N33/58G01N33/532
CPCG01N33/558G01N33/573G01N33/577G01N33/58G01N33/532G01N2333/91182
Inventor 王金斌曾海娟杨倩雯
Owner SHANGHAI ACAD OF AGRI SCI
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