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Indirect competitive ELISA kit for detecting malachite green in aquatic product

A detection kit, colorless malachite green technology, applied in the biological field, can solve the problems of high cost, slow speed, low cost, etc., and achieve the effect of low cost, good safety, and image of result judgment

Inactive Publication Date: 2006-05-03
SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C +1
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  • Claims
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AI Technical Summary

Problems solved by technology

Chromatography (HPLC) detection technology is mainly used to detect MG at home and abroad. Although it is sensitive and quantitatively accurate, it is slow in large-scale detection and the cost is relatively high.
The rapid screening method represented by competitive enzyme-linked immunosorbent assay (ELISA) is the mainstream technology of immunology. It is fast, low-cost, and can be tested in large quantities. There is no report on this technology at home and abroad.

Method used

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  • Indirect competitive ELISA kit for detecting malachite green in aquatic product
  • Indirect competitive ELISA kit for detecting malachite green in aquatic product

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Embodiment Construction

[0024] According to technical scheme of the present invention, concrete preparation is as follows:

[0025] (1) Preparation of colorless malachite green microporous detection plate

[0026] Use 0.2M carbonate buffer at pH 9.6 as the coating solution, dilute the colorless malachite green and albumin conjugate (LMG-RSA or LMG-OVA) to 2 μg / ml, add poly In a styrene microwell plate, coat overnight at 4°C, spin dry, add 0.01M pH7.4 phosphate buffer containing 1% gelatin at 200 μl / well to block at 37°C for 2 hours, wash and dry, then add 20% The sucrose phosphate buffer solution was protected at room temperature for 3 hours, dried in a drying room, and stored in a packaging bag containing a desiccant.

[0027] (2) Preparation of sample diluent, washing solution, monoclonal antibody dilution, and stop solution

[0028] Sample or serum diluent is 0.01M phosphate buffer (KH 2 PO 4 0.2g, Na 2 HPO 4 12H 2 O 2.9g, NaCl 8g, dilute to 1000ml, pH7.4, add 0.5ml Tween-20); 10× concentra...

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Abstract

The invention relates to a colorless green malachite green indirect compete ELISA test agent box of an aquatic product. The test plate of the agent box is a split 96 holes enzyme mark plate which coats colorless green malachite green and albumin coupling material; the anti-colorless green malachite green antibody is a monoclonal antibody which is prepared by the colorless green malachite green and the albumin coupling material immune mouse; the tested sample uses ethyl hexoate and cyclohexane to extract after uniform and adjusts PH value for detecting. The quoting standard of the sample detecting is that it is a regression curve which uses the logarithm value of the sample density as abscissa and uses the degradation rate as ordinate. We could read the density of the corresponding sample from the degradation rate of each sample. The sensibility of the test method is 0.0023ª–g / ml; the test range is 0.0016-1ª–g / ml.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a rapid diagnosis method and apparatus for a kind of medicine of aquatic meat food, in particular to malachite green indirect competition ELISA detection kit Background technique [0002] Colorless malachite green (Leucomalachite green, LMG) is the main metabolic residue of malachite green (MG) in organisms, which has a carcinogenic risk to humans. [0003] Malachite green, also known as aniline green, Victoria green or Chinese green, is a green crystal with metallic luster, soluble in water, ethanol and methanol, and the color of the aqueous solution is blue-green (the maximum absorption wavelength is 618nm). It was first introduced in 1913 In 2000, it was discovered that dyes such as malachite green can destroy pathogenic bacteria without causing damage to the host, so that these dyes can be used as antiseptics, trypanocidal drugs and other medical effects. Since the emergence of su...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/15G01N33/531G01N33/543
Inventor 郭德华李健王权龚朋飞薛飞群顾惠明陈永军陈燕军
Owner SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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