Immunofluorescence detection method for measuring virus carried rate of vector insects

A technology for immunofluorescence detection and mediator insects, which is applied in the direction of fluorescence/phosphorescence, measuring devices, and material analysis through optical means, can solve the problems of not being suitable for large-scale sample detection, high technical requirements, and high detection costs, and achieve shortening The detection time, the result is intuitive and accurate, and the effect of short time consumption

Inactive Publication Date: 2014-07-23
FUJIAN AGRI & FORESTRY UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

Although these techniques can be used for the detection of vector insects in the field, they all have some shortcomings, such as high detection cost, high technical requirements, time-consuming detection, and not suitable for large-scale sample detection, etc.

Method used

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  • Immunofluorescence detection method for measuring virus carried rate of vector insects
  • Immunofluorescence detection method for measuring virus carried rate of vector insects
  • Immunofluorescence detection method for measuring virus carried rate of vector insects

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Embodiment Construction

[0033] 1. Reagent preparation

[0034] 1) 0.01mol / L PBS: Weigh 8 g NaCl, 0.2 g KCl, 1.44 g Na 2 HPO 4 and 0.24 g KH 2 PO 4 , dissolved in 800 ml of distilled water, adjusted the pH value to 7.2 with HCl, and added distilled water to make the volume to 1L.

[0035] 2) Fixative solution: Weigh 4 g of paraformaldehyde and dissolve it in 100 ml of 0.01 mol / L PBS buffer solution, heat and stir at 50°C-60°C to dissolve, and store at 4°C.

[0036] 3) Permeate: Add 0.2 mL of TritonX-100 solution to 10 mL of 0.01 mol / L PBS buffer, stir well to dissolve, and store at 4°C.

[0037] 4) Antibody diluent: Weigh 0.3g BSA into a 15ml centrifuge tube, add 10mL of 0.01 mol / L PBS buffer, stir well to dissolve, and store at 4°C.

[0038] 5) Anti-fading solution: Measure 90 ml of glycerol and 10 mL of 0.01 mol / L PBS buffer solution, mix well, and store at 4°C.

[0039] 6) Fluorescent antibody: For southern rice black-streaked dwarf virus (SRBSDV), firstly, the IgG of the polyclonal antibod...

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Abstract

The invention discloses an immunofluorescence detection method for rapidly measuring the virus carried rate of vector insects on a large scale. The method comprises the steps of classifying the vector insects collected in the field, dissecting, fixing, permeating, preparing antibody labeling liquid, incubating antibody, carrying out slide preparation, observing, and calculating the number of insects carrying virus by statistics to obtain the virus carried rate of the vector insects. The method is low in detection cost, only consumes four hours, is visual and accurate in detection result and high in sensitivity, and can be used for detecting the virus carried rate of the vector insects in the field on a large scale.

Description

technical field [0001] The invention belongs to the field of agricultural disease detection, and in particular relates to an immunofluorescence detection method for large-volume and rapid determination of vector insect toxin-carrying rate. Background technique [0002] Rice is the largest food crop in my country. However, rice diseases caused by rice viruses have occurred in large areas in southern my country in recent years. It has become the most serious virus disease in rice production and seriously threatens my country's food production. [0003] At present, the main rice viruses are Southern rice black streaked dwarf virus (SRBSDV), rice black streaked dwarf virus ( Rice black streaked dwarf virus , RBSDV), rice dwarf virus ( Rice dwarf virus , RDV), rice dwarf virus ( Rice ragged stunt virus , RRSV), rice tumor dwarf virus ( Rice gall dwarf virus , RGDV) and rice stripe virus ( Rice stripe virus , RSV), these rice-infecting viruses are transmitted by vector ins...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569G01N21/64
CPCG01N33/56983G01N21/6458G01N33/533
Inventor 贾东升魏太云陈红燕吴维陈倩毛倩卓
Owner FUJIAN AGRI & FORESTRY UNIV
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