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Sorghum mosaic virus tri-anti sandwich enzyme-linked immuno sorbent assay kit

A sorghum mosaic virus, enzyme-linked immunosorbent technology, applied in the direction of antiviral immunoglobulin, serum immunoglobulin, measuring device, etc., can solve the problems of low specificity and accuracy, achieve high repeatability, Strong specificity and high detection effect

Inactive Publication Date: 2010-10-06
云南省农业科学院生物技术与种质资源研究所
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Problems solved by technology

At present, it is reported that the detection of sorghum mosaic virus is mainly based on electron microscope observation and RT-PCR detection. These methods have the disadvantages of specificity and low accuracy.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0051] 1. Isolation and purification of sorghum mosaic virus

[0052] Samples of typical sugarcane diseased plants were collected in the field, which were detected by electron microscopy and PCR to contain a large amount of SrMV, and did not contain other viruses. After collection, non-toxic sugarcane plants were inoculated. After the samples containing SrMV produced typical mosaic symptoms, the leaves were collected, and the veins were removed. Store in a -80°C freezer for later use.

[0053] The purification steps and conditions of sorghum mosaic virus are:

[0054] (1) Add PB extraction buffer to the susceptible leaves frozen at -80°C, the weight-to-volume ratio of diseased leaves to PB extraction buffer is 1:2, and the unit is g / ml; the PB extraction buffer contains 0.2mol / L phosphate buffer, phosphate buffer containing 0.01mol / L EDTA-Na and 0.1% mercaptoethanol, pH7.6;

[0055] (2) Fully homogenate with an electric mixer, filter with double-layer gauze;

[0056] (3) Ad...

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PUM

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Abstract

The invention discloses a sorghum mosaic virus tri-anti sandwich enzyme-linked immuno sorbent assay (TAS-ELISA) kit which comprises positive control, negative control, a sorghum mosaic virus polyclonal antibody, a sorghum mosaic virus monoclonal antibody, an enzyme labeled antibody and other materials and drugs, wherein the sorghum mosaic virus polyclonal antibody is obtained through rabbit immune separation serum after separating and purifying a sorghum mosaic virus sugarcane separator; and the sorghum mosaic virus monoclonal antibody is obtained by mouse immune, cell fusion and clone purification. The TAS-ELISA kit prepared by the invention has high detection sensitivity, strong specificity, high repeatability and high economic usefulness.

Description

technical field [0001] The invention relates to a sorghum mosaic virus triple-antibody sandwich enzyme-linked immunosorbent detection kit and a preparation method thereof. Background technique [0002] Sorghum mosaic virus, SrMV, is a member of the Potatovirus family Potatoviridae. The virion is 750nm long, has single-stranded positive-sense RNA, and the molecular weight of the coat protein is 36kDa. It forms pinwheel-shaped endosomes in susceptible tissues. Sugarcane mosaic disease caused by sorghum mosaic virus is common and serious in Zhejiang and Yunnan of my country. Infected plants have shortened internodes, decreased sugar content, and degraded quality. The virus can be transmitted with sugarcane and aphids, and has a high risk of outbreak. It is one of the most harmful virus diseases in sugarcane production. Therefore, it is necessary to establish a fast, accurate and sensitive detection method. At present, the detection of sorghum mosaic virus is mainly based on ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531C12N7/02C07K16/10C07K16/06
Inventor 张仲凯董家红苏晓霞丁铭方琦李婷婷张丽珍
Owner 云南省农业科学院生物技术与种质资源研究所
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