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Antibody chip for detecting compositae plant virus diseases, and kit and method of antibody chip

A technology of antibody chips and Compositae plants, which is applied in the direction of measuring devices, scientific instruments, instruments, etc., can solve the problems of long process, expensive manpower, material and financial resources, etc., and achieve the effect of simple equipment, saving manpower and convenient operation

Active Publication Date: 2014-10-22
兰州海关技术中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the widely used detection methods of plant viruses include enzyme-linked immunosorbent assay (Enzyme Linked Immuno Sorbent Assay, ELISA) and reverse transcription-polymerase chain amplification (Reverse Transcription-PCR, RT-PCR), but conventional methods Detecting multiple pathogens in a sample is a long process that requires a lot of manpower, material and financial resources

Method used

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  • Antibody chip for detecting compositae plant virus diseases, and kit and method of antibody chip
  • Antibody chip for detecting compositae plant virus diseases, and kit and method of antibody chip
  • Antibody chip for detecting compositae plant virus diseases, and kit and method of antibody chip

Examples

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Effect test

Embodiment 1

[0029] Example 1: see figure 1 , an antibody chip applied to detection of Compositae plant virus disease, characterized in that it includes a basement membrane and specific capture antibodies (primary antibodies) for 7 kinds of viruses that are harmful to Compositae plants that are separately spotted on the basement membrane, The capture antibody (primary antibody) of described 7 kinds of viruses: anti-bean yellow mosaic virus, anti-Arabidopsis mosaic virus, anti-bean wilt virus, anti-cucumber mosaic virus, anti-tobacco crisp virus, anti- -tobacco ringspot virus, anti-tomato spotted wilt virus; 3 positive quality control points P are also encapsulated on the basement membrane, and the spots are made of alkaline phosphatase-labeled IgG and 3 negative quality control points N are encapsulated. Prepare carbonate buffer solution at pH 9.2-9.8.

Embodiment 2

[0030] Embodiment 2: the preparation method of the described antibody chip that is applied to detection Compositae plant virus disease, its steps are:

[0031] (1) Antibody dilution: dilute 7 kinds of polyclonal capture antibodies (primary antibodies) with the capture antibody (primary antibody) dilution buffer, and the concentration of the diluted capture antibody (primary antibody) is 1 / 18-1 / 3 times of the original antibody solution , namely BYMV, ArMV, BBWV, CMV, TRV, TRSV and TSWV, the antibody concentration used for spotting is 1:17, 1:14, 1:17, 1:14, 1:15, 1:3 of the original antibody solution and 1:15 times, and at the same time, add 1:10 diluted alkaline phosphatase-labeled donkey anti-goat IgG stock solution on the chip as the positive quality control point of the chip, and the chip spotting solution as the negative quality control point; the spotting buffer is: carbonic acid Sodium Na 2 CO 3 1.59g, sodium bicarbonate NaHCO 3 2.93g, sodium azide NaN 3 0.2g, double...

Embodiment 3

[0035] Embodiment 3: a kind of antibody chip kit that is applied to detection Asteraceae plant virus disease, its main feature is, comprise antibody chip; 7 kinds of alkaline phosphatase-labeled detection antibody (secondary antibody) corresponding to virus: anti -Bean yellow mosaic virus, anti-Arabidopsis mosaic virus, anti-bean wilt virus, anti-cucumber mosaic virus, anti-tobacco crisp virus, anti-tobacco ringspot virus, anti-tomato spotted wilt virus; Asteraceae Positive controls for plant viruses, including positive controls for Bean Yellow Mosaic Virus, positive controls for Arabidopsis Mosaic Virus, positive controls for Bean Wilt Virus, positive controls for Cucumber Mosaic Virus, and positive controls for Tobacco Crisp Virus Positive control, positive control of tobacco ringspot virus, positive control of tomato spotted wilt virus; alkaline phosphatase; detection antibody (secondary antibody) dilution buffer; washing buffer PBST; alkaline phosphatase chromogenic substra...

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Abstract

The invention relates to an antibody chip for screening and diagnosing compositae plant virus diseases, and a kit and a method of the antibody chip. The antibody chip for screening and diagnosing compositae plant virus diseases is mainly characterized by comprising a base membrane and specific capture antibodies (primary antibodies) which are respectively pointed on the base membrane and used for capturing seven types of viruses with harm to the compositae plant, wherein the antibodies (primary antibodies) for capturing seven types of viruses have resistance to bean yellow mosaic virus, resistance to arabis mosaic virus, resistance to curtobacterium flaccumfaciens virus, resistance to cucumber mosaic virus, resistance to tobacco rattle virus, resistance to tobacco ringspot virus and resistance to tomato spotted wilt virus; three positive quality control points P, IgG pointed to be alkaline phosphatase mark, three negative quality control points N and a carbonate buffer solution pointed to reach a pH being 9.2-9.8 are also encapsulated on the base membrane. The antibody chip for screening and diagnosing compositae plant virus diseases has the advantages that the antibody chip for detecting a plurality of viruses of the compositae plant is successfully built, so that comprehensive detection is carried out on a plurality of pathogens at a high flux.

Description

technical field [0001] The invention relates to an antibody chip, a kit and a method for screening and diagnosing plant virus diseases, mainly for bean yellow mosaic virus, Arabidopsis mosaic virus, bean wilting virus and cucumber mosaic virus of Compositae plants Seven kinds of viruses, tobacco crisp virus, tobacco ringspot virus and tomato spotted wilt virus, belong to the field of biotechnology. Background technique [0002] There are a large number of medicinal, ornamental and economic plants in Compositae. Virus diseases are one of the main limiting factors for the production of Compositae. Bean yellow mosaic virus (Bean yellow mosaic virus, BYMV), Arabis mosaic virus (Arabis mosaic virus , ArMV), broad bean wilt virus (BBWV), cucumber mosaic virus (CMV), tobacco rattle virus (Tobacco rattle virus, TRV), tobacco ringspot virus (Tobacco ringspot virus, TRSV) ) and tomato spotted wilt virus (TSWV) can infect Compositae plants. BYMV infects a variety of plants such as As...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983
Inventor 文朝慧王军平周小平李儒赵政
Owner 兰州海关技术中心
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