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Immunochromatographic assay test strip for bacterial fruit blotch of melons

A technology for immunochromatographic detection and fruit spot disease, which is applied in measurement devices, analytical materials, instruments, etc., can solve problems such as difficulty in grass-roots production and promotion, high experimental technical requirements, and low detection accuracy, and achieve intuitive and accurate results. , the effect of a wide range of applications and broad market prospects

Inactive Publication Date: 2015-07-15
河南省农业科学院园艺研究所
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Domestic melon bacterial fruit spot detection methods mainly include serological detection and molecular detection. Serological detection mainly includes ELISA method, which has the disadvantages of low detection accuracy and high false positives. PCR, etc., its advantages are sensitive, accurate, and fast, but its disadvantage is that it requires high experimental technology, and requires specific equipment and professional technical personnel for technical equipment.
Although the above methods have their own advantages, they also have obvious defects, and it is difficult to promote them in grassroots production.

Method used

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  • Immunochromatographic assay test strip for bacterial fruit blotch of melons
  • Immunochromatographic assay test strip for bacterial fruit blotch of melons

Examples

Experimental program
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Effect test

preparation example Construction

[0028] 2. Preparation of anti-bacterial fruit spot monoclonal antibody

[0029] Immunize 8-week-old Balb / c mice with cultured bacterial fruit spot bacteria at a dose of 20 μg / mouse. After the first immunization, each booster immunization interval is 2 weeks. Subcutaneous and intraperitoneal injections, and the third booster immunization is 2 weeks later , ELISA determination of serum antibody titers. After the last booster immunization, the interval was more than 4 weeks, and 4 days before cell fusion, 50 μg of immune antigen was injected into the tail vein and abdominal cavity, and diluted with sterilized PBS to a volume of 200 μL.

[0030] On the 3rd day after hyperimmunization of spare mice for cell fusion, the eyeballs were pulled out and bled, and the serum was collected as a positive control. Mice were killed by neck breaking, and the body surface was disinfected with 75% alcohol. In an ultra-clean bench, open the abdominal cavity to take out the mouse spleen, and wash...

Embodiment 1

[0048] A kind of melon bacterial fruit spot disease immunochromatographic detection test strip, see figure 1 , figure 2 , including the support bottom plate 4 placed at the bottom, providing the entire product skeleton, sequentially connecting the sample buffer layer 1 arranged on the support bottom plate 4, the carrier layer 5 covered with labeled protein, the coating adsorption layer 2 and the liquid absorption layer 3, and a test line 7 and a quality control line 6 are provided on the coating adsorption layer 2 .

[0049] The sample buffer layer 1 is a glass fiber membrane, the coated adsorption layer 2 is made of nitrocellulose membrane, the liquid absorbing layer 3 is made of absorbent paper, and the support base 4 is made of rigid PVC coated with glue on one side. The carrier layer 5 is made of polyester film, which is sprayed with quantum dot-labeled anti-bacterial fruit spot monoclonal antibody IgG, the quality control line 6 is made of rabbit anti-mouse IgG, and th...

Embodiment 2

[0054] A kind of melon bacterial fruit spot disease immunochromatographic detection test strip, see figure 1 , figure 2 , including the support bottom plate 4 placed at the bottom, providing the entire product skeleton, sequentially connecting the sample buffer layer 1 arranged on the support bottom plate 4, the carrier layer 5 covered with labeled protein, the coating adsorption layer 2 and the liquid absorption layer 3, and a test line 7 and a quality control line 6 are provided on the coating adsorption layer 2 .

[0055]The sample buffer layer 1 is a glass fiber membrane, the coated adsorption layer 2 is made of nitrocellulose membrane, the liquid absorbing layer 3 is made of absorbent paper, and the support base 4 is made of rigid PVC coated with glue on one side. The carrying layer 5 is made of polyester film, on which the rabbit anti-bacterial fruit spot polyclonal antibody IgG labeled with quantum dots is sprayed, the quality control line 6 is made of goat anti-rabb...

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Abstract

The invention specifically relates to an immunochromatographic assay test strip for bacterial fruit blotch of melons. The immunochromatographic assay test strip for the bacterial fruit blotch of melons comprises a support bottom plate arranged on the lowest layer, as well as a sample buffer layer, a bearing layer loading labelled proteins, a coating adsorption layer and a liquid absorption layer, which are sequentially connected and arranged on the support bottom plate, and at least one test line and quality control line is formed on the coating adsorption layer. The immunochromatographic assay test strip for the bacterial fruit blotch of melons is high in specificity, rapid, simple and convenient, wide in application range, convenient to popularize and apply, and capable of being widely used in basic areas.

Description

technical field [0001] The invention relates to a device for detecting bacterial fruit spot disease of melons, in particular to an immunochromatographic detection test strip for bacterial fruit spot disease of melons. Background technique [0002] The pathogen of bacterial fruit spot disease of melons is Acidovorax avenae subsp.citrulli, the pathogen is short rod-shaped, Gram stain negative, does not produce fluorescence, is strictly aerobic, and has a single polar flagella ; Can grow at 41°C, but not at 4°C; On KB medium, it presents milky white, round, smooth, entire, raised, opaque colonies with a diameter of 1-2mm. The results of tobacco allergic reaction caused by the bacteria were inconsistent, arginine hydrolase was not produced, gelatin liquefaction was weak, and oxidase and 2-ketogluconic acid tests were positive. The pathogen can attach to the surface of melon seeds, and may also invade the inner tissue of the seeds, and survive in the surface layer of the endospe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/558
CPCG01N33/56911G01N33/558
Inventor 李晓慧赵卫星常高正梁慎徐小利康利允
Owner 河南省农业科学院园艺研究所
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