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Rapid detection kit for cucumber green mottle mosaic viruses

A green mottled flower and leaf virus technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of limited wide application, unsatisfactory sensitivity of colloidal gold immunochromatography test strips, etc., and achieve the effect of improving detection efficiency

Inactive Publication Date: 2014-02-05
曹冬梅 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the sensitivity of colloidal gold immunochromatographic test strips is not ideal, which greatly limits its wide application in the detection field.

Method used

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  • Rapid detection kit for cucumber green mottle mosaic viruses
  • Rapid detection kit for cucumber green mottle mosaic viruses
  • Rapid detection kit for cucumber green mottle mosaic viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1. Preparation of Cucumber Green Mottled Mosaic Virus Rapid Detection Kit and Cucumber Green Mottled Mosaic Virus Rapid Detection Test Paper

[0026] 1. Preparation of fluorescent nanoparticle-cucumber green mottle mosaic virus antibody conjugate reaction solution

[0027]Take 50 μL (10 mg / ml) of fluorescent particles (purchased from Shenzhen Yirui Biotechnology Co., Ltd.), add 550 μL 50 mM MES (pH 6.0), 200 μL each of 100 mg / mL sulfo-NHS and EDC, shake at room temperature for 30 min; centrifuge, discard Supernatant; resuspend particles with 1 mL of 50 mM MES (pH 6.0); add appropriate amount of monoclonal antibody to cucumber green mottle mosaic virus (purchased from Beijing Lanbury Biotechnology Co., Ltd.), shake at room temperature for 1 h; add an equal volume of PBS -BSA (BSA content 2%, pH7.4), continue to react for 1h; centrifuge, discard the supernatant; wash the particles with 50mM MES (pH6.0) 3 times, 1mL / time; resuspend with 300μL (10mM Tris-HCl (pH8.0...

Embodiment 2

[0048] Embodiment 2. Sample detection

[0049] 1. Test sample preparation

[0050] Test sample preparation: Take 1g of the leaves to be tested in a clean plastic bag, add 3ml of PBST (1×PBS, 0.5% Tween20, pH7.4), rub the leaves in the plastic bag, and take the juice as the test sample for testing.

[0051] 2. Use the Cucumber Green Mottle Mosaic Virus Rapid Detection Kit to detect samples

[0052] Add the prepared test sample to the reaction tank 12 of the quick detection box for cucumber green mottle mosaic virus prepared in Example 1.3, add fluorescent nanoparticles-cucumber green mottle mosaic virus antibody conjugate reaction solution, blow and mix ;Insert the test strip, incubate at 40°C for 3 minutes, and observe the chromatography results with naked eyes under ultraviolet light after 5 minutes.

[0053] If a reaction plate with holes is used for detection, directly add the test sample into the blind well 12 of the reaction plate with holes 11, blow and mix well; inser...

Embodiment 3

[0061] Embodiment 3. Contrast test with colloidal gold immunochromatography

[0062] Monoclonal antibody-colloidal gold conjugates were prepared by referring to the method in the literature "Development of Pumpkin Mosaic Virus Colloidal Gold Immunochromatographic Test Strips" ([J]. Phytosanitary, 2012, 38(5):88-91.), And refer to the method of Example 1 to prepare cucumber green mottled mosaic virus colloidal gold immunochromatography detection box and detection test paper. Colloidal gold immunochromatography detection was carried out with reference to the method of Example 2.

[0063] 1. Detection sensitivity comparison test

[0064] Use the test form of embodiment 2.2 (detection box, reagent bottle) to compare the quick detection test strip of cucumber green mottled mosaic virus prepared by the embodiment of the present invention 1.3 and the colloidal gold immunochromatographic detection test strip of cucumber green mottled mosaic virus sensitivity.

[0065] The same posi...

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Abstract

The invention discloses a rapid detection kit for cucumber green mottle mosaic viruses. The rapid detection kit comprises a kit cover, a kit body and a test paper strip, wherein the box cover is provided with a reaction groove for containing a fluorescent nano particle-cucumber green mottle mosaic virus antibody conjugate; the kit body is internally provided with a test paper strip storing position; the test paper strip is a rapid detection test paper strip for the cucumber green mottle mosaic viruses and is composed of a bottom plate, and a sample cushion, a chromatography film and a water absorption cushion which are attached on the bottom plate and are closely connected in sequence; the chromatography film is provided with a detection region and a quality control region; the detection region is used for fixing a monoclonal antibody or a polyclonal antibody which is subjected to specific reaction with the cucumber green mottle mosaic viruses. The detection kit provided by the invention can be used for finishing the detection of the cucumber green mottle mosaic viruses of plants in very short time, has good specificity and sensitivity and is an idea cucumber green mottle mosaic virus infection detection tool.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a device for detecting plant cucumber green mottle mosaic virus infection by using immune chromatography technology. Background technique [0002] Cucumber green mottle mosaic virus is one of the important phytosanitary viruses, which can cause crop yield loss up to 15%, posing a serious threat to the production of Cucurbitaceae crops. In 2005, watermelons in Gaizhou City, Liaoning Province were infected with cucumber green mottle mosaic virus, causing heavy losses. Since then, outbreaks have been discovered in Guangxi, Liaoning, Hebei, Shandong, Guangdong, Beijing and other places, which have seriously threatened the production of watermelons, melons, cucumbers and other crops. Therefore, it is extremely important to strengthen the detection of the virus. [0003] Preventing the entry and spread of seeds carrying cucumber green mottle mosaic virus from endemic areas is the p...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/533
CPCG01N33/56983G01N33/533G01N33/558G01N33/577
Inventor 曹冬梅李鑫曹际娟李立梅
Owner 曹冬梅
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