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Dot enzyme-linked immunosorbent assay (ELISA) method for detecting bemisia tabaci carrying tomato yellow leaf curl virus and application of method

A technology of tomato yellowing curved leaves and bemisia tabaci, applied in the biological field, can solve the problems that serological methods cannot be applied to the detection of vector insects, the inability to clarify the poisoning situation of single bemisia tabaci, and strong dependence on instruments, etc., to achieve Reliable detection results, low cost and high sensitivity

Active Publication Date: 2013-02-20
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows researchers to quickly test if bamboo saproves carrying certain types of disease are infected without having physical contact or knowledge about them. It also includes methods that make this testing process faster than traditional laboratory tests while maintaining good reliability over multiple sample sizes. Additionally, there're no expensive equipment needed but only one device needs to work properly. Overall, these technical features improve efficiency and effectiveness during emergency situations such as pandemics caused by Yellow Leaf Curl (BSCCO), which affect people worldwide causing severe damage through spreading diseases like malaria.

Problems solved by technology

This patented technical problem addressed in this patents relates to identifying mistranscribed areas where tomatoyellow leaf curler syndrome occurs more frequently than other diseases such as bamboo yellows that are commonly found throughout Japan today without any signs indicating symptoms like these diseases.

Method used

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  • Dot enzyme-linked immunosorbent assay (ELISA) method for detecting bemisia tabaci carrying tomato yellow leaf curl virus and application of method
  • Dot enzyme-linked immunosorbent assay (ELISA) method for detecting bemisia tabaci carrying tomato yellow leaf curl virus and application of method
  • Dot enzyme-linked immunosorbent assay (ELISA) method for detecting bemisia tabaci carrying tomato yellow leaf curl virus and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: Detection of poisonous rate of Bemisia tabaci samples in the field

[0059] The present invention utilizes the method of dot-ELISA to detect the carrying situation of tomato yellow leaf curl virus by whitefly in the field, calculates the virus carrying rate of whitefly, predicts the outbreak trend of local tomato yellow leaf curl virus, and takes preventive measures in advance , to serve agricultural production.

[0060] 110 Bemisia tabaci were collected from the field of the Vegetable Institute of Hangzhou Academy of Agricultural Sciences with an insect extractor ( figure 1 ) and stored in a 4°C refrigerator for a short period of time. After removing Bemisia tabaci from the refrigerator at 4°C, place it in a petri dish and wash it with ddH 2 O soaking, dipped the tip of a gun to pick up a single head of Bemisia tabaci, placed it on a clean parafilm, and dried it at room temperature. Draw a line with a pencil on the outer paper of the nitrocellulose membr...

Embodiment 2

[0061] Example 2: Development of a dot-ELISA kit for detecting tomato yellow leaf curl virus carried by Bemisia tabaci

[0062] 1. Detection of tomato yellow leaf curl virus (TYLCV) dot-ELISA kit ( Figure 4 ) contains the following materials and reagents:

[0063] 1) Kit instruction manual 1 copy

[0064] 2) Reagents and materials:

[0065] 0.05 mol / L PBS 10ml

[0066] TYLCV monoclonal antibody 0.1 ml

[0067] HRP-labeled goat anti-mouse IgG secondary antibody (A4416 Sigma) 0.1 ml

[0068] TMB chromogenic substrate solution (W4121 Promega) 10 ml

[0069] 10× concentrated blocking solution (0.1 mol / L PBST) 10 ml

[0070] 10× Concentrated Antibody Diluent (50% skimmed milk powder, 0.1mol / L PBST) 20 ml

[0071] 20× concentrated washing solution (0.2 mol / L PBST) 60 ml

[0072] Blocking agent (skimmed milk powder) 15 g

[0073] NC film (Amersham) 5 sheets

[0074] 2. The kit instruction manual includes:

[0075] 2.1 Principle of the kit:

[0076] dot-ELISA is an ELISA ...

Embodiment 3

[0111] Example 3: Using a kit to detect Bemisia tabaci in the field

[0112] The kit can be used to detect the virus-carrying rate of Bemisia tabaci samples in the field, and thousands of Bemisia tabaci can be detected at the same time. The results are fast, reliable, high-throughput and easy to operate. It is used to investigate the situation of tomato yellow leaf curl virus carried by Bemisia tabaci on a large scale in the field, and to predict the outbreak of the disease in time. Figure 6 It is part of the results of using the kit to detect Bemisia tabaci in Hangzhou field vegetables.

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Abstract

The invention discloses a dot enzyme-linked immunosorbent assay (ELISA) method for detecting bemisia tabaci carrying a tomato yellow leaf curl virus and application of the method. Field bemisia tabaci is collected, pretreated and grinded, monoclonal antibody of the tomato yellow leaf curl virus is used, the dot-ELISA method which is used for detecting virus transmission medium bemisia tabaci is established, a quick kit is developed and researched, and the tomato yellow leaf curl virus which is carried by the bemisia tabaci is detected. By the aid of the method, the virus carrying rate of the field bemisia tabaci can be surveyed in a large scale, and the field tomato yellow leaf curl disease outbreak trend is predicted. The method is quick, efficient, sensitive, special, high in flux and simple to operate. The tomato yellow leaf curl disease can be diagnosed quickly, predicted and controlled scientifically.

Description

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Claims

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Application Information

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Owner ZHEJIANG UNIV
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