Paralytic shellfish-poison competitive enzyme-lined immue quantitative detecting reagent box, its preparing and use

An enzyme-linked immunosorbent assay and quantitative detection technology, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as complex matrix, achieve high repeatability, perfect acute poisoning treatment system, and good specificity.

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 ​​

Method used

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  • Paralytic shellfish-poison competitive enzyme-lined immue quantitative detecting reagent box, its preparing and use
  • Paralytic shellfish-poison competitive enzyme-lined immue quantitative detecting reagent box, its preparing and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Preparation of sample antigen: Weigh the sample, wash it, homogenate it, boil it with 10% hydrochloric acid, centrifuge it, and take the supernatant.

Embodiment 2

[0026] Example 2: Preparation of monoclonal antibody and detection of monoclonal antibody properties

[0027] 1. Dissolve 25 μg of PSP shellfish poison (STX) in 25 μL of dimethyl sulfoxide (DMSO), add bovine serum albumin (BSA), carbodiimide (EDC), N-hydroxysulfosuccinimide (Sulfo-NHS) were dissolved in phosphate buffered saline (PBS) at pH 7.4, respectively. The above four substances were mixed together at a ratio of 1:3:1:1, and stirred overnight at 4°C. The next day, transfer the reaction solution into an ultrafiltration tube, centrifuge at 6,000 rpm for 80 min at 4°C, wash with PBS solution and collect the STX-BSA conjugate on the ultrafiltration membrane, and freeze it at -20°C for future use. Conjugates as immunogens.

[0028] 2. Select 3 8-week-old female Balb / c mice, weighing 18-22 g, and dilute 600 μL (1 mg / mL) of the aliquoted antigen to 1.5 mL with normal saline. Add an equal volume of Freund's complete adjuvant, and use a syringe to repeatedly suck and stir in t...

Embodiment 3

[0034] Example 3: Determination of Molecular Weight of Monoclonal Antibody

[0035] 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 separation, the separation gel was fixed, stained, decolorized and dried (Molecular Cloning Test Guide 2nd Edition, 1999). In SDS-PAGE discontinuous electrophoresis, the relative mobility Mr of each protein (the ratio of the distance the protein migrates from the origin...

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PUM

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Abstract

The invention relates to batching detecting kit for paralysis bel toxin compete enzyme coupling immune that the feature is that it includes enzyme marking board, paralysis 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 paralysis 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 a kit for quantitative detection of paralytic shellfish poisoning by enzyme-linked immunosorbent assay (ELISA) and its preparation and application, in particular to a method for quantitatively detecting paralytic shellfish poisoning by using a competitive enzyme-linked immunosorbent assay (Enzyme Linked Immunosorbent Assay, ELISA). Kit of shellfish toxin and its preparation and application. technical background [0002] Paralytic shellfish poisoning (PSP) is a potential neurotoxin produced in a variety of algae and accumulates in shellfish through the food chain. Paralytic shellfish poisoning (PSP) causes respiratory paralysis, leading to death in 8% of cases, and blocks sodium ion channels in nerve and muscle septa. The lethal dose is 1-3mg, and the clinical manifestations of paralytic shellfish poisoning (PSP) ingestion of 0.5-1.0ug are numbness and respiratory disturbance. PSP is composed of saxitoxin (STX) and its...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/543G01N33/535
Inventor 曹际娟周卫东
Owner 曹际娟
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