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Method for detecting patulin and special enzyme linked immunosorbent assay kit thereof

An enzyme-linked immunosorbent reagent, patulin technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as difficult to achieve rapid detection of batch samples

Inactive Publication Date: 2011-06-01
北京维德维康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The detection methods of patulin mainly include thin layer chromatography (TLC), high performance liquid chromatography (HPLC) and gas chromatography (GC), but the above methods are difficult to achieve rapid detection of batch samples

Method used

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  • Method for detecting patulin and special enzyme linked immunosorbent assay kit thereof
  • Method for detecting patulin and special enzyme linked immunosorbent assay kit thereof
  • Method for detecting patulin and special enzyme linked immunosorbent assay kit thereof

Examples

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Effect test

Embodiment 1

[0059] Embodiment 1, patulin hapten, patulin antigen and patulin antibody and their preparation

[0060] 1. Structure and preparation of patulin hapten

[0061] Patulin was purchased from Sigma Aldrich Company, the product catalog number is 32759;

[0062] Dissolve patulin, glutaric anhydride and 4-dimethylaminopyridine (DMAP) (molar ratio 1:18:5) in anhydrous tetrahydrofuran, shake and react at 37°C for 3 hours, add distilled water to terminate the reaction, and obtain the compound of formula I as Patulin hapten (PAT-HG).

[0063]

[0064] 2. Patulin Antigen and Its Preparation

[0065] 1. Synthesis of patulin immune antigen

[0066] Patulin is a small molecule chemical substance without immunogenicity. To prepare anti-patulin antibodies, it must be coupled to a protein carrier with a relatively large molecular weight (such as BSA, OVA, etc.). Due to the simple structure of patulin, the characteristic group in the molecular structure is more prominent after a spacer ar...

Embodiment 2

[0092] Example 2, ELISA kit for detecting patulin and its preparation and application

[0093] 1. The enzyme-linked immunosorbent assay kit consists of the following materials:

[0094] 1. The patulin coater shown in formula III described in Example 1;

[0095] 2. ELISA plate;

[0096] 3. Patulin monoclonal antibody: secreted by the patulin monoclonal antibody hybridoma cell line PAT with the deposit number CGMCC No.3392;

[0097] 4. Standard patulin: the concentrations of the standard solutions are 0 μg / L, 1.0 μg / L, 5.0 μg / L, 20.0 μg / L, 50.0 μg / L and 100.0 μg / L; the standard patulin was purchased from Sigma Aldrich company; catalog number 32759; dilute to the above concentrations with sample diluent;

[0098] 5. Goat anti-mouse secondary antibody labeled with horseradish peroxidase;

[0099] 6. Substrate chromogenic solution: composed of liquid A and liquid B, liquid A is an aqueous solution of 2% carbamide peroxide, liquid B is an aqueous solution of 1% tetramethylbenzidin...

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Abstract

The invention discloses a method for detecting patulin and a special enzyme linked immunosorbent assay kit thereof. The enzyme linked immunosorbent assay kit comprises patulin monoclonal antibodies. The patulin monoclonal antibodies are secreted by patulin monoclonal antibody hybridoma cell strains PAT with collection number CGMCC No.3392. The enzyme linked immunosorbent assay kit qualitatively or quantitatively detects the patulin content of fruits or juice by mainly adopting an indirect competitive ELISA method. The kit and the detection method have low pretreatment requirement for a sample and simple sample pretreatment process, and can quickly detect a large batch of samples at the same time; and by adopting the high-specificity patulin monoclonal antibodies, the detection method is convenient and feasible and has the characteristics of high specificity, high sensitivity, high precision, high accuracy and the like. The kit plays a great role in the detection of the patulin.

Description

technical field [0001] The invention relates to a method for detecting patulin and a special ELISA kit. Background technique [0002] Patulin (PAT) is mainly produced by some fungi (Penicillium expanses, Penicillium clavulae, and etc.) produced as a secondary metabolite. Molecular formula is C 7 h 6 o 4 , melting point 100 ° C, molecular weight 154, is a colorless needle crystal, soluble in water, ethanol, acetone, ethyl acetate and chloroform. Glister was first discovered and isolated and purified at Oxford University in 1941. In the early days, patulin was considered as a broad-spectrum antibiotic that could be used for the treatment of human colds, but subsequent animal experiments found that it had strong acute and subacute toxicity, immunotoxicity, teratogenicity, and mutagenicity to experimental animals. Since then, the focus of research on patulin has shifted to its toxicity and pollution to food and feed. Patulin mainly contaminates apples, hawthorns and their...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/566G01N33/543
Inventor 江海洋吴小平王战辉史为民徐飞王照鹏王进米铁军潘净如
Owner 北京维德维康生物技术有限公司
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