Rice stripe virus monoclonal antibody and rice stripe virus immunodetecting method

A technology of rice streak virus and monoclonal antibody, applied in the field of microbiology, can solve the problems of unsuitable large-scale detection of samples and high cost

Inactive Publication Date: 2005-12-14
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the methods used to detect the infection rate of SBPH in the world include ELISA, RT-PCR, Northern hybridization, etc., but RT-PCR and Northern hybridization are expensive and not suitable for large-scale detection of samples; and domestic RSV monoclonal antibodies have not been prepared for use Virus detection, and ELISA detection with monoclonal antibody is fast, sensitive, simple, and suitable for large-scale detection. The establishment of this method will help our grassroots detection personnel to carry out early detection and timely warning of poisonous insects in the early stage

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The steps of the indirect method are:

[0052] 1) For coating samples, use 0.05M Na for rice samples to be inspected 2 CO 3 -NaHCO 3 Dilute the coating solution at 1:30, add 100ul to each hole at 37°C for 2 hours or overnight at 4°C;

[0053] 2) Sealing: 1% skimmed milk powder-PBST 37 degrees closed for 30 minutes;

[0054] 3) Join our company to develop monoclonal antibody against rice stripe virus specific cell strain (3B9), dilute this monoclonal antibody ascites as 50,000, and add 100ul per well to react at 37°C for 1 hour;

[0055] 4) Washing: Wash the ELISA reaction plate 3 times with pH 7.2, 0.01M PBST, each time for 3 minutes;

[0056] 5) Add horseradish peroxidase-labeled goat or rabbit anti-mouse IgG, horseradish peroxidase-labeled goat anti-mouse is sold by SIGMA or other companies, and add working concentration of horseradish peroxidase-labeled goat anti-mouse , Add 100ul to each hole at 37°C for 1 hour;

[0057] 6) Washing: same as step 4;

[0058] 7) Color dev...

Embodiment 2

[0066] Except that the test object is an insect sample, and the test object---insect sample is diluted 1:300, the other operations are the same as in Example 1.

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Abstract

The present invention discloses monoclonal antibody of rice stripe virus (RSV) and its immunodetecting method. Specific monoclonal antibody of RSV is used as tool in establishing immunodetecting method for RSV in rice and RSV propagating insect. The specific process includes: immunizing bal b/c mouse with purified RSV to prepare hybridoma cell, specifically screening hybridoma with positive culture supernatant to obtain four monoclonal antibody strains of specifically resisting RSV, and establishing direct/indirect ELISA detection method with these monoclonal antibodies. The establishment and application of the said method makes it possible to detect rice disease sample and insect with RSV conveniently, accurately and fast.

Description

Technical field [0001] The invention relates to a monoclonal antibody of rice stripe virus and its agricultural application-an immunological detection method of rice stripe virus, which belongs to the technical field of microbiology. Background technique [0002] Rice stripe virus (RSV) caused rice stripe leaf blight first discovered in Kanto, Japan in 1897, and later occurred in North Korea, Ukraine and China. Since the virus was discovered in Jiangsu and Zhejiang in 1962, it has been circulating in 16 provinces, municipalities and autonomous regions across the country. [2] . The disease is still very common in Yunnan, Liaoning, Beijing, Henan, Shandong, Jiangsu and Shanghai, especially Baoshan, Chuxiong and Kunming in Yunnan, Shuangqiao in Beijing, Yuanyang in Henan, Jining in Shandong and Jiangyan and Hong in northern Jiangsu. It is more common in Ze and other places. [0003] The main reasons for its continued outbreak are: the warm winter climate in recent years is conducive...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08
Inventor 周益军陈正贤程兆榜周雪平王桂珍
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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