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A kit and method for detecting Tilletia dwarf

A technology of Tilletia dwarf and a kit, which is applied in the field of kits for detecting Tilletia dwarf, can solve problems such as failure to separate

Active Publication Date: 2017-11-07
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

In 2004, Gao Qiang (Gao Qiang. Molecular detection of Tilletia tritici smut [D]. Changsha, Hunan Agricultural University, 2004) used RAPD technology to find a line that could isolate Tilletia tritici strains and Tilletia tritici strains Different from the bands of other smut strains, but unfortunately no different bands were found between Tilletia dwarf and Tilletia erythroderma, and the two could not be separated
[0004] The enzyme-linked immunoassay method has the advantages of fast speed and good specificity, and is widely used in the detection of pathogenic bacteria. However, the specificity and sensitivity of this technology depend on the specific recognition and capture performance between the antibody-antigen used. And some factors that affect the color development of colloidal gold

Method used

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  • A kit and method for detecting Tilletia dwarf
  • A kit and method for detecting Tilletia dwarf
  • A kit and method for detecting Tilletia dwarf

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1. Preparation of colloidal gold test paper for detecting Tilletia vulgaris

[0044] Composition of colloidal gold test paper for detection of Tilletia vulgaris

[0045] 1. Micropore reagent

[0046] There is a micro-hole plug on the micro-hole; the bottom of the micro-hole is freeze-dried with Tilletia dwarf monoclonal antibody-colloidal gold marker, the amount of freeze-drying is 50μl;

[0047] Two, test strips ( figure 1 ):

[0048] The test strip is composed of a bottom plate, a sample absorption pad, a reaction film, a water absorption pad, and a protective film;

[0049] The sample absorbent pad 1, the reaction film 2, the absorbent pad 3, and the protective film are sequentially pasted on the bottom plate 6 in order. The end of the sample absorbent pad is connected to the reaction membrane, and the end of the reaction membrane is connected to the absorbent pad. The beginning of the pad is aligned with the beginning of the bottom plate, and the end of the absorbent ...

Embodiment 2

[0090] Example 2. Detection limit and specificity test of Tilletia vulgaris colloidal gold test paper

[0091] (1) Detection limit test

[0092] The standard Tilletia vulgaris (TCK1 teliospore) was diluted to 0, 5, 10, 20 mg / L, and the dilution used was a phosphate buffer with a pH of 7.2 and 0.2 mol / L.

[0093] Use colloidal gold test paper for testing. Add 200μl of sample to the microwell reagent each time, mix well, and after incubating for 5 minutes, insert the test paper into the test. The result is: when 0, 5mg / L Tilletia dwarf is tested, the quality control area of ​​the test strip shows color , The detection area does not show color and is negative; when 10, 20mg / L Tilletia vulgaris is tested, the quality control area and detection area of ​​the test strip are both colored and positive; it shows that the test strip is positive for wheat dwarf The detection limit of Tilletia is 10mg / L.

[0094] (2) Specificity test

[0095] Specificity is often expressed by cross-reaction rate...

Embodiment 3

[0096] Example 3. Application of Tilletia dwarf colloidal gold test paper

[0097] 1. Use test paper to detect Tilletia vulgaris

[0098] The test strip can detect Tilletia vulgaris. The detection method is as follows:

[0099] 1. Detection method

[0100] Add dropwise the sample solution (PBS buffer and cosolvent) to be tested to the microwell reagent. After mixing, incubate for 5 minutes. Insert the MAX line-labeled end of the test paper into the microwell reagent down and watch the results within 5 minutes.

[0101] 2. Judgment of test results

[0102] When the concentration of T. dwarf in the sample is higher than or equal to 10 mg / L, the colloidal gold antibody binds to T. dwarf, and a red band appears in the test area due to the sandwich reaction of the double antibody, which is positive. When the concentration of T. wheat dwarf bacteria in the sample is lower than 10 mg / L, there is no red band in the detection area due to the lack of antibody antigen reaction during the detectio...

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Abstract

The invention "a kit and method for detecting Tilletia dwarfis" belongs to the detection technology of plant pathogens. Including a test strip and a microporous plate, the test strip includes a bottom plate, a sample absorbent pad, a reaction film and a water absorbent pad; the sample absorbent pad, reaction film, and water absorbent pad are sequentially pasted and installed on the bottom plate; its features In that: there are freeze-dried micropore reagents in the microwells of the micropore plate, and the micropore reagents refer to gold-labeled antibodies obtained by labeling the monoclonal antibody described in claim 2 with colloidal gold; the reaction membrane is divided into A detection area and a quality control area, the detection area is coated with Tilletia tritici polyclonal antibody, and the quality control area is coated with anti-antibody. The kit of the invention has good specificity and very low detection limit, and is suitable for the prevention and control or quarantine of T. tritici.

Description

Technical field [0001] The invention relates to a plant pathogen detection technology, in particular to a kit and a method for detecting Tilletia triticina. technical background [0002] Wheat dwarf smut caused by Tilletia controversa Kühn (TCK) is an important international quarantine disease, which has a devastating hazard to wheat production. It is also one of the main species of biological invasion research in our country. 1. (Wang Yuan. Tilletia dwarf[M]. Selection of Entry Plant Quarantine Pests in China, edited by Animal and Plant Quarantine Bureau of the People’s Republic of China and Plant Quarantine Laboratory of the Ministry of Agriculture, Beijing: China Agriculture Press, 1997, 95 ~98). In the years when wheat dwarf bunt is prevalent, the yield loss caused by it is generally 20-50%, and it can reach 75-90% in severe cases, or even no production. The pathogen is highly resistant to stress, its teliospores can survive in the soil for 3-7 years, and the teliospores en...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/14G01N33/577G01N33/569
Inventor 高利陈万权刘太国刘博
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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