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Colloidal gold test strip for detection of Tilletia controversa Kuhn

A technology of Tilletia dwarf and colloidal gold test paper, which is applied in the field of colloidal gold test strips for the detection of Tilletia dwarf, can solve the problems of cumbersome steps, time-consuming and laborious, etc., and achieve good specificity, high titer, strong affinity effect

Inactive Publication Date: 2015-09-23
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the method is cumbersome and takes one day from sampling to obtaining results, which is time-consuming and laborious, and needs to be carried out in the laboratory

Method used

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  • Colloidal gold test strip for detection of Tilletia controversa Kuhn
  • Colloidal gold test strip for detection of Tilletia controversa Kuhn

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1, acquisition of Tilletia triticum monoclonal antibody

[0037] 1. Preparation of hybridoma cells

[0038] The experimental animals were BALB / c mice, provided by the Chinese Academy of Military Medical Sciences.

[0039] 1. Antigen preparation

[0040] Using T. tritici spores as the immunogen, the mice were immunized according to the routine mouse immunization method.

[0041] 2. Preparation of hybridoma cells

[0042] (1) Select mice with better serum detection of wheat dwarf smut and perform booster immunization. At the same time, prepare SP2 / 0 myeloma cells, culture them in a 37°C carbon dioxide incubator, and perform fusion after 3 days of booster immunization.

[0043] (2) BALB / c mice immunized with T. tritici were killed by cervical dislocation and soaked in 75% alcohol for 5 minutes. Take the mouse out to dry, put it in an ultra-clean workbench, cut off the epithelium to expose the epithelium of the abdominal cavity, cut a small opening in the epi...

Embodiment 2

[0067] Embodiment 2, be used to detect the preparation of the colloidal gold test strip of Tilletia tritici

[0068] According to the preparation method and process of colloidal gold test strips, corresponding auxiliary materials are prepared, which mainly include: NC film, PS bottom plate, gold standard pad, sample pad, absorbent paper (the above auxiliary materials are all in accordance with the existing standards of Beijing Qinbang Biotechnology Co., Ltd. Procurement and configuration of small molecule detection process).

[0069] (1) Preparation of gold particles

[0070] 1. Reagents:

[0071] 2% HAuCL4 (sigma) solution; the dilution method is double distilled water (the resistivity is about 18.2).

[0072] 3% trisodium citrate solution (sigma), which needs to be freshly prepared, and then filtered through a 0.35um membrane filter.

[0073] Instrument: electromagnetic heating stirring jacket thermostat + whirling flow round flask + snake-shaped condenser tube + stirring...

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Abstract

The invention provides a colloidal gold test strip for detection of Tilletia controversa Kuhn. The colloidal gold test strip comprises a sample pad, a colloidal gold marking pad, an NC film, absorbent paper and a PS base plate, and the sample pad, the colloidal gold marking pad, the NC film and the absorbent paper are sequentially arranged on the PS base plate along a sample chromatography direction. A colloidal gold marked monoclonal antibody resistant to Tilletia controversa Kuhn covers the colloidal gold marking pad, the NC film is provided with a detection line and a quality control line sequentially along the sample chromatography direction, the detection line is coated by inactivated Tilletia controversa Kuhn spores, and the quality control line is coated with GaMIgG (goat-anti-mouse IgG serum). The monoclonal antibody resistant to Tilletia controversa Kuhn is secreted by a hybridoma cell with the preservation number of CGMC No.10883. By adoption of the colloidal gold test strip for detection, no instrument or equipment is required, only 15 minutes are taken from material acquisition to detection result acquisition, and simplicity and convenience in operation and accurate results are realized.

Description

technical field [0001] The invention relates to the field of prevention and treatment of Tilletia dwarf, in particular to a colloidal gold test strip for detecting Tilletia tritici. Background technique [0002] Tilletia controversa Kühn (TCK) is an important international quarantine disease caused by Tilletia controversa Kühn (TCK), and it is also a kind of dangerous plant disease in foreign quarantine in my country. The disease is very harmful and difficult to control. Once introduced, it will cause devastating damage to wheat production, and the incidence rate is usually approximately equal to the loss rate of wheat yield. According to Hoffmann's report, in popular years, the loss rate caused by TCK is generally 20% to 50%, and the highest can reach 75% to 90%. In addition to yield loss, pollination of infested wheat decreases with infection severity and quality is greatly affected by the presence of TCK teliospores. T. dwarf is a potential crisis to my country's wheat ...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 高利蔚慧欣沈慧敏刘太国刘博陈万权
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI