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Test paper for detecting anthrax bacillus and colloidal gold of spore thereof, preparation method and application thereof

A technology of Bacillus anthracis and detection test paper, which can be applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of long time consumption, complicated and complicated operation, etc., and achieve the effect of saving manpower and material resources.

Inactive Publication Date: 2009-07-29
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing immunological detection methods often use capsular antibodies as enzyme labels or fluorescent antibody staining for diagnosis. In molecular biology, there are PCR methods, and there are many traditional inspection methods, including bacterial isolation and culture, hemolytic experiments, and capsular staining. , phage lysis experiment, beading experiment, etc., but the above method is time-consuming, and a large amount of purified bacteria must be carried out, requiring the purchase of expensive instruments, cumbersome and complicated operations, and requires professionals with skilled operating skills to accurately detect

Method used

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  • Test paper for detecting anthrax bacillus and colloidal gold of spore thereof, preparation method and application thereof
  • Test paper for detecting anthrax bacillus and colloidal gold of spore thereof, preparation method and application thereof
  • Test paper for detecting anthrax bacillus and colloidal gold of spore thereof, preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, detect the preparation of the colloidal gold test paper of bacillus anthracis

[0047] 1. Preparation of colloidal gold probes

[0048] (1) HAuCl 4 It is first prepared as a 0.01% aqueous solution, and 100ml of pure water is taken and heated to boiling. Accurately add 1ml of 1% HAuCl under magnetic stirring 4 , while adding 1.5ml mass fraction of 1% trisodium citrate aqueous solution. After the color is stable, continue to heat and boil for 15 minutes. After cooling to room temperature, add pure water to make up to 100ml, and store in the dark at 4°C.

[0049] (2)K 2 CO 3 Adjust the PH value to about 8.2-8.5.

[0050] (3) Adjust the colloidal gold to a suitable pH, preferably 8.0-8.5, more preferably 8.2, and dilute the antibody to 0.2 mg / ml with 5 mM PB (pH 7.2).

[0051] 2. Preparation of gold standard pad

[0052] Take 1ml / tube of colloidal gold with pH 8.2 and distribute it in 1.5ml centrifuge tubes. Add the optimal labeling dose of antibody t...

Embodiment 2

[0056] Embodiment 2, Bacillus anthracis and its spore detection test paper

[0057] Referring to Fig. 1, the reaction support is a 6.2cm × 0.4cm PCV plate; the absorbent pad is a 2cm × 0.4cm oil filter paper; the nitrocellulose membrane of 1.8cm × 0.4cm is successively coated with rabbit anti-sheep IgG (purchased from Dingguo Biotechnology company, 1mg / ml), goat anti-St polyclonal antibody 1.5mg / ml (containing 1% BSA blocking agent), containing 0.4cm×0.4cm colloidal gold-labeled goat anti-St polyclonal antibody (gold concentration: 11ug PB diluted ; Optimum PH value is 8.5) glass fiber membrane; Sample pad is the glass fiber membrane of 2.7cm * 0.4cm; Promptly formed Bacillus anthracis and its spore detection test paper.

Embodiment 3

[0058] Embodiment 3, detection method

[0059] See attached figure 2 , mix the sample to be tested with the sample diluent, and then use a pipette to add the sample mixture to the sample hole of the test paper. The liquid in the sample goes up by siphon action, and the result is interpreted in 10-15 minutes:

[0060] If it contains Bacillus anthracis antigen, it will form a corresponding complex with the colloidal gold-labeled goat anti-St polyclonal antibody on the test paper, and combine with the flow rabbit anti-St polyclonal antibody coated on the nitrocellulose membrane to form a red line. A red band forms at the "T".

[0061] Regardless of whether it contains the corresponding antigen or not, the colloidal gold-labeled polyclonal antibody continues to crawl upward and forms a red precipitation line with the rabbit anti-goat IgG coated on the membrane, that is, a red band is formed at "C". This line is a quality control line. If the colloidal gold fails, this line will ...

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Abstract

The invention discloses a piece of detection test paper which comprises (1) a reaction support; (2) an absorbent pad; (3) a nitric acid fiber membrane which is coated with a detection strip of a bacillus anthracis antibody and a quality control strip of a quality control antibody; and (4) a GAB gold-labelled antibody protective film which comprises the bacillus anthracis antibody marked with colloidal gold. The test paper of the invention can be coordinated with small instrument for semi-quantitative detection, does not need professional training, has clear and easily-identified result, can objectively preserve data, operates simply, is easy for promotion, can be suitable for the grass-roots units, the on-site detection of unexpected events, and the investigation of epidemiology, and plays a supporting role for the infection diagnosis of the bacillus anthracis.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a test paper for detecting bacillus anthracis antigen and spores thereof, a preparation method thereof and an application in detecting bacillus anthracis. Background technique [0002] Anthrax is a zoonotic acute infectious disease caused by Bacillus anthracis. People have known this disease and struggled with it for more than 100 years, and have accumulated rich experience in prevention and treatment. However, at the present stage, anthrax still poses a threat to human beings, with frequent outbreaks around the world, especially in developing countries. In recent years, the most serious human epidemic in Africa has affected more than 10,000 people. A large number of livestock die every year, causing major economic losses, and human anthrax cases continue to occur. [0003] Humans get sick through direct or indirect contact with infected animals, mainly by stripp...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/543G01N33/532
Inventor 王静张晓龙孙肖红李伟杨宇胡孔新谢士嘉
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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