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Amnestic shellfish-poison competitive immue quantitative detection reagent box, its preparation and use

An enzyme-linked immunosorbent immunoassay and quantitative detection technology, which is applied in the field of amnestic shellfish poison competitive enzyme-linked immunosorbent immunoassay kits, can solve the problems of complex matrix and achieve high repeatability, good specificity, and high sensitivity

Inactive Publication Date: 2007-06-13
曹际娟 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Shellfish toxin residues in food (aquatic products) are very small, generally at the level of μg / kg, and the matrix is ​​complex. It is often a challenge for analytical chemists to analyze and identify certain trace components in this mixture

Method used

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  • Amnestic shellfish-poison competitive immue quantitative detection reagent box, its preparation and use
  • Amnestic shellfish-poison competitive immue quantitative detection reagent box, its preparation and use

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1: Preparation of sample antigen: remove the shell, wash the shellfish with double distilled water, and homogenize with a homogenizer; add 70mL of 50% methanol solution to 10.0g sample, and stir for 5min; centrifuge at 3000g for 10min; take 100μL The supernatant was diluted 1:10 with double distilled water (1+9); take 50 μL for analysis with the kit, and the dilution factor at this time is 80;

Embodiment 2

[0028] Example 2: Preparation of monoclonal antibody and detection of monoclonal antibody properties

[0029] After dissolving 10 grams of ASP (DA) samples in 300 milliliters of pyridine according to the mixed anhydride method, add 50 milliliters of succinic anhydride, stir until the reaction is complete, wash with acid water, filter the precipitate with suction, and dry to obtain a crude product. The crude product was dissolved in acetone, mixed with an appropriate amount of silica gel, dried in the air, separated by column chromatography on a silica gel column, monitored by TLC, and the points of Rf=0.09 (developing solvent, petroleum ether:acetone=3:1) were collected and combined, Dry at room temperature to obtain activated chondroitin (DA).

[0030] Dissolve 50 mg of the above substance in 5 mL of dioxane, place at 4°C for 10 min, add 25 µL of tri-n-butylamine, mix well, then add 19 µL of isobutyl chloroformate, stir magnetically at 4°C for 30 min, and this is liquid A. A...

Embodiment 3

[0037] Example 3: Determination of Molecular Weight of Monoclonal Antibody

[0038] In the Bio-Rad electrophoresis tank, pour 12% separation gel, and add a small amount of water, so that the edge of the gel is horizontal after solidification. The filter paper was sucked to remove water, poured into the electrode buffer, and electrophoresed at a constant current (I=12mA) for 10min. After aspirating the electrode buffer, pour 3% stacking gel into the tank. After gelation, 10 μl of purified monoclonal antibody and standard protein samples were added to each sample well. The sample was concentrated in the stacking gel for 20min at a constant flow (I=20mA), and then separated in the separating gel for 2.5h. After the separation, the separating gel was fixed, dyed, decolorized and dried (the second edition of Molecular Depression Test Guidelines, 1999). In SDS-PAGE discontinuous electrophoresis, the relative mobility Mr of each protein (the ratio of the distance the protein migra...

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Abstract

The invention relates to batching detecting kit for oblivion bel toxin compete enzyme coupling immune that the feature is that it includes enzyme marking board, oblivion conch toxin standard antigen liquid, covering liquid, sealing liquid, horse radish peroxide enzyme labeling antigen bonder, substrate, drying out liquid and cleaning solution. It mainly makes the enzyme marking board. It is used to test the oblivion conch toxin and has high sensitivity, good specificity, high repeatability and rapid testing. It decreases testing cost, expands sample monitoring quantity and effectively controls biotoxin in food. It has important effect in rapid identifying, testing and curing in toxicant.

Description

technical field [0001] The present invention relates to amnestic shellfish poison competitive enzyme-linked immunosorbent quantitative detection kit and its preparation and application, specifically relates to a kind of using competitive enzyme immunoassay (Enzyme Linked Immunosorbent Assay, ELISA) to quantitatively detect amnestic shellfish product Kit of shellfish toxin and its preparation and application. technical background [0002] Amnesiac Shellfish Toxin (ASP) is represented by Domoic Acid (DA) in its chemical structure. After ingestion, it can produce nausea, vomiting, abdominal pain, diarrhea, etc., and at the same time have dizziness, coma and other symptoms similar to neurotoxicity. In severe cases, it is a general term for marine biological toxic substances existing in shellfish that cause permanent loss of part of memory. At present, the food safety standard generally adopted in the world (fresh, frozen or canned oysters, clams and mussels food) is that the sa...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/532
Inventor 曹际娟周卫东
Owner 曹际娟
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