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Aflatoxin molecule early warning method for agricultural products

A technology for aflatoxin and agricultural products, which is applied in the field of aflatoxin molecular early warning of agricultural products, can solve problems such as insufficiency and lack of molecular early warning research examples, and achieve the effects of fast early warning, promoting high-quality development, and promoting food safety.

Active Publication Date: 2021-11-12
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there is still no example of molecular early warning research before microbial toxin contamination such as aflatoxin at home and abroad, and it is difficult to meet the urgent needs of early warning

Method used

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  • Aflatoxin molecule early warning method for agricultural products
  • Aflatoxin molecule early warning method for agricultural products
  • Aflatoxin molecule early warning method for agricultural products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 Preparation of aflatoxin early warning molecule AFT-YJFZ01 that can act as a standard substance

[0034] According to the following formula configuration Cha's medium: 3% (w / v) sucrose, 0.3% (w / v) NaNO3, 0.1% (w / v) K2HPO4, 0.05% (w / v) MgSO4 7H2O, 0.05% ( w / v) KCl, 0.001% (w / v) FeSO4, pH 6.5, configured to obtain Chapei's medium. Random selection of public literature "Distribution, Toxicity and Infection of Aspergillus flavus in Typical Peanut Production Areas in China" - Chinese Academy of Agricultural Sciences master's degree thesis, author Zhang Xing, page 33 - published toxin-producing fungal strain HLJ-1 , HeNZY-2, HuBha-24, JXZS-29-2, LNct-6, GXfc-34, GDZJ-108-19, JSnt-1, HuNdx-7, HBHA-8-17 and other 10 strains were inoculated into the above-mentioned In Cha's medium, culture at 28°C 200rpm / min for 5 days, fully homogenize and break the cells by conventional methods, and then use conventional protein purification systems, protein electrophoresis, immu...

Embodiment 2

[0053] The above-mentioned aflatoxin early warning molecule AFT-YJFZ01 antibody can also be obtained by using all or part of the peptides of the aflatoxin early warning molecule AFT-YJFZ01 after knowing the entire sequence of the antibody preparation process. Example 2 Nanobody Preparation of Aflatoxin Early Warning Molecule AFT-YJFZ01

[0054] Using AFT-YJFZ01 as the immune antigen, immunizing alpaca or Balb / c mice in a conventional way, and then using the known conventional nanobody or mouse monoclonal antibody preparation technology, it can be developed and obtained.

[0055] Dissolve the AFT-YJFZ01 prepared above in conventional PBS buffer or normal saline to a concentration of not less than 0.1mg / mL, then mix and emulsify with Freund's complete adjuvant in equal volume, and inject it subcutaneously or intradermally at multiple points on the back Immunize the alpaca, and then boost the immunization once every 2-4 weeks, and replace the Freund's complete adjuvant with Freun...

Embodiment 3

[0057] Example 3 Monoclonal Antibody Preparation of Aflatoxin Early Warning Molecule AFT-YJFZ01

[0058] Using AFT-YJFZ01 as the immune antigen, immunizing alpaca or Balb / c mice in a conventional way, and then using the known conventional nanobody or mouse monoclonal antibody preparation technology, it can be developed and obtained.

[0059] Dissolve the AFT-YJFZ01 prepared above in conventional PBS buffer or normal saline to a concentration of not less than 0.1 mg / mL, then mix and emulsify with Freund's complete adjuvant in equal volume, and inject it subcutaneously or intradermally at multiple points on the back BALB / c mice were then boosted once every 2-4 weeks, and Freund's complete adjuvant was replaced with Freund's incomplete adjuvant during the booster immunization. Use conventional ELISA process to monitor the immune effect, and after the serum titer of BALB / c mice no longer rises, then isolate the splenocytes of immunized mice, splenocytes are fused with mouse myelom...

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Abstract

The invention relates to an aflatoxin molecule early warning method for agricultural products. The aflatoxin early-warning molecule concentration in the agricultural product sample is determined, when the determined aflatoxin early-warning molecule concentration is smaller than 0.2 microgram / g, the aflatoxin pollution level is 0.2 microgram / g or below, and the risk is low; when the determined aflatoxin early warning molecule concentration is greater than 1.2 microgram g / g, the aflatoxin pollution level is more than 10 microgram g / kg, the risk is high; when the measured concentration of the aflatoxin early-warning molecule is 0.2 microgram / g to 1.2 microgram / g, the risk is medium, the aflatoxin early-warning molecule is AFT-YJFZ01 peptide, and the amino acid sequence of the aflatoxin early-warning molecule is shown as SEQ ID NO. 1. The method is used for early warning the risk that agricultural products are polluted by aflatoxin, and is high in speed, high in timeliness and practicability and easy to popularize and apply.

Description

technical field [0001] The invention relates to an early warning method for aflatoxin molecules in agricultural products. Background technique [0002] Aflatoxins are highly toxic and harmful, and are the most common pollutants that contaminate food. In recent years, there has been an overall trend of aggravating pollution, seriously threatening food safety and people's health. Aflatoxins are the most toxic mycotoxins in nature, among which aflatoxin B1 is a Class I carcinogen identified by the International Agency for Research on Cancer (IARC), which has caused many poisoning incidents of human and livestock groups. It has become one of the main reasons for the high incidence of liver cancer cases. According to the statistics retrieved by Web of Science in the past 5 years: there are more than 110 types of food and raw materials contaminated by aflatoxin, ranking first among pollutants. However, so far, there is still no research example of molecular early warning before ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543
CPCG01N33/535G01N33/543G01N2333/38Y02A50/30
Inventor 张奇唐晓倩李培武白艺珍姜俊
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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