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Aflatoxin

The Severity of Food Contamination Caused by Aflatoxin

Aflatoxin (AFT) is a bifuran ring toxin produced by strains such as Aspergillus flavus and Aspergillus parasiticus Speare. About 20 kinds of its derivatives are named B1, B2, G1, G2, M1, M2, GM, P1, Q1, and poison alcohol. In 1993, aflatoxin was classified as a carcinogen by the World Health Organization (WHO) Cancer Research Institute. Among them, B1 is the most toxic and carcinogenic. It can induce liver cancer in animals and can cause acute poisoning and death to certain animals. Aflatoxin and its producing bacteria are widely distributed in nature and can be detected in seeds and processed products of cereals and oil crops, dried and fresh fruits. AFT can be transferred to the milk, liver, kidney, and muscle tissue of animals through food accumulate. People who accidentally eat food contaminated with aflatoxin will be poisoned. The pollution of aflatoxin in food spreads to all regions of the world. Generally speaking, food pollution in tropical and subtropical regions is more serious.

The Necessity of Monitoring Aflatoxin Contamination with ELISA

Fig. 1 The chemical structures of Aflatoxin B1

There are many types of AFT, and the basic structure contains a difuran ring and naphthalene. The former is its toxic structure, and the latter may be related to its carcinogenesis. AFT is hardly soluble in water, and resistant to high temperatures. Generally, heat treatment will cause little damage to it, and it will only decompose at the melting point. Human consumption of food contaminated by AFT can cause acute poisoning, causing liver necrosis and bleeding, and even liver cancer. At the same time, raising livestock and poultry with contaminated feed will reduce the productivity of livestock and poultry, slow down weight gain, and indirectly cause major harm to humans. The AFT poisoning incident in India in 1974 was the most serious, resulting in 397 poisonings and 106 deaths. AFT poses a great threat to food safety. In recent years, methods for detecting AFT, especially ELISA, have been rapidly developed.

ELISA Types

Direct competitive ELISA, Indirect competitive ELISA

The Advantages of ELISA Testing

Can quickly and easily detect aflatoxin
High-throughput detection and analysis aflatoxin
High specificity and sensitivity to aflatoxin

ELISA Procedure for Aflatoxin Testing

The microplates were coated with 100 μL of protein G (25 µg/mL) in bicarbonate buffer (0.1M, pH 8.6) at 4°C overnight and then blocked with 1 mg/mL of BSA solution at 37°C for 2 h.

After washing three times with PBST (pH 7.4), 100 µL of anti-AFB1 mAbs (0.8 ng/mL) was added and incubated at 37°C for 1 h.

After washing with PBST three times, 50 μL of solution (2.5 μg/mL) and 50 μL of sample solution were added into each plate well, incubation at 37°C for 1 h.

After washing with PBST three times and ultrapure water twice. Then 100 µL of glucose solution was added to each well reacting at 37°C for 1 h.

After washing, 40 µL of the AuNRs and 30 µL HRP (100 µg/mL) was added to each well.

The absorbance was measured at 650 nm by microplate reader.

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References

XIONG, Y.; et al. Plasmonic ELISA based on enzyme-assisted etching of Au nanorods for the highly sensitive detection of aflatoxin B1 in corn samples. Sensors and Actuators B: Chemical. 2018, 267: 320-327.
ZHANG, Xiya.; et al. Development of a new broad-specific monoclonal antibody with uniform affinity for aflatoxins and magnetic beads-based enzymatic immunoassay. Food Control. 2017, 79: 309-316.

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