High-flux enzyme linked immunosorbent assay kit for aflatoxin M1 in milk and detection method of kit

An enzyme-linked immunosorbent reagent and aflatoxin technology, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of high non-specific adsorption and difficult long-term storage of kits, and achieve the effect of improving specificity and accuracy

Pending Publication Date: 2018-07-06
CHINA NAT CENT FOR FOOD SAFETY RISK ASSESSMENT +1
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Based on this, it is necessary to provide a high-pass method for aflatoxin M1 in milk to solve the problem that traditional kits are not easy to store for a long time, and the non-specific adsorption in the enzyme-linked immunosorbent assay for aflatoxin M1 is still high. Quantitative enzyme-linked immunosorbent assay kit and detection method thereof

Method used

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  • High-flux enzyme linked immunosorbent assay kit for aflatoxin M1 in milk and detection method of kit
  • High-flux enzyme linked immunosorbent assay kit for aflatoxin M1 in milk and detection method of kit
  • High-flux enzyme linked immunosorbent assay kit for aflatoxin M1 in milk and detection method of kit

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preparation example Construction

[0067] The preparation method of described blocking solution, comprises the following steps:

[0068] The recipe amount of Na 2 HPO 4 12H 2 O, NaH 2P o 4 2H 2 0 is dissolved in an appropriate amount of water to prepare the first buffer solution; according to the formula, sucrose, skim milk powder, the first buffer solution, and a protein stabilizer are dissolved in an appropriate amount of water to obtain the first solution;

[0069] Specifically, add sucrose, skimmed milk powder, the first buffer solution, and protein stabilizer to an appropriate amount of water to dissolve first, so as to form a buffered pH solution condition, and then add fetal bovine serum to avoid the destruction of fetal bovine serum components. Further, finally add Proclin300 has anti-corrosion effect and can improve the anti-corrosion performance of the blocking solution.

[0070] As an optional embodiment, before adding the fetal calf serum and Proclin300 of the formula amount to the first solut...

Embodiment 1

[0097]1.1. Preparation of blocking solution

[0098] Weigh 15.0g of sucrose, 9.0g of skimmed milk powder, Na 2 HPO 4 12H2O 5.8g, NaH 2 PO 4 2H 2 0.6g, 3.0g of protein stabilizer AEP-HBC were placed in a 1L glass container, added 500mL of pure water and stirred at room temperature with a magnetic stirrer for 30min to dissolve, then added 10mL of glycerin and 450mL of pure water and continued to stir with a magnetic stirrer for 12h, then Add 40 mL of fetal bovine serum and 300 μL of Proclin300, dilute to 1 L and stir for 30 min with a magnetic stirrer to obtain a blocking solution.

[0099] 1.2. Preparation of Aflatoxin M1 Artificial Antigen Conjugate

[0100] Weigh 20mg of aflatoxin M1 and 10mg of maleic anhydride respectively into a conical flask, add 20mL of pyridine, and heat at reflux at 70°C for 6h. Use a rotary evaporator to evaporate the pyridine in the mixture, extract with ethyl acetate and remove water with anhydrous sodium sulfate, and dry in vacuum for 12 hour...

Embodiment 2

[0144] The blocking solution was prepared as follows, and other preparation methods were the same as in Example 1.

[0145] Weigh 12.0g of sucrose, 7.2g of skimmed milk powder, Na 2 HPO 4 12H 2 O 4.64g, NaH 2 PO 4 2H 2 0.48g, protein stabilizer AEP-HBC 2.4g are placed in a 1L glass container, add 500mL pure water and stir at room temperature with a magnetic stirrer for 30min to dissolve, then add 8mL of glycerin and 450mL pure water and continue to stir with a magnetic stirrer for 12h, then Add 32 mL of fetal bovine serum and 240 μL of Proclin300, dilute to 1 L and stir for 30 min with a magnetic stirrer to prepare a blocking solution.

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Abstract

The invention provides a high-flux enzyme linked immunosorbent assay kit for aflatoxin M1 in milk and a detection method of the kit. The kit comprises a closing solution, an enzyme-labeled antibody concentrated solution, an enzyme label dilution solution, a ELISA plate coated with coating antigens and a substrate developing liquid, wherein the closing solution contains fetal calf serum, skim milkpowder and a protein stabilizer, each 1000mL of the closing solution contains 32.0mL-48.0mL of the fetal calf serum, 7.2g-10.8g of the skim milk powder and 2.4g-3.6g of the protein stabilizer; and theenzyme label dilution solution contains enzyme-labeled antibodies, fetal calf serum and Proclin300, and each 1000mL of the enzyme label dilution solution contains 50mL-100mL of the enzyme-labeled antibodies and 100mu L-400mu L of Proclin300.

Description

technical field [0001] The invention relates to the technical field of mycotoxin detection in food, in particular to a high-throughput ELISA kit for aflatoxin M1 in milk and a detection method for aflatoxin M1 in milk. Background technique [0002] Aflatoxin M is a group of structurally related toxic compounds. There are two main types of aflatoxin in milk, M1 and M2. In 1993, aflatoxin was classified as a class 1 carcinogen by the World Health Organization. It is a highly toxic and highly toxic substance. It is harmful to the liver tissue of humans and animals, and can cause liver cancer or even death in severe cases. [0003] There are currently three main methods for detecting aflatoxin M1: [0004] 1. Biological detection method. Including seed germination test, vomiting test, etc., which are not conducive to rapid detection and are rarely used. [0005] Second, the physical and chemical detection method. Including thin layer chromatography (TLC) and high performance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 骆鹏杰李敬光赵云峰陈霞蒋双勤王华丽刘卿
Owner CHINA NAT CENT FOR FOOD SAFETY RISK ASSESSMENT
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