Fluorescence quantitative detection card for aflatoxin B1 and application thereof

A technology for fluorescent quantitative detection and aflatoxin, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of high technical level requirements, expensive equipment, cumbersome processing, etc., avoid separation and washing steps, and be easy to operate and portable strong effect

Pending Publication Date: 2019-10-18
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

High performance liquid chromatography has reliable results and high sensitivity, but the equipment is expensive, the technical level is high, the sample pretreatment is cumbersome and time-consuming, and it is not suitable for rapid and on-site detection
Thin-layer chromatography, limited accuracy, not quantitative
Enzyme-linked immunosorbent assay and colloidal gold immunochromatography are fast and convenient, easy to operate, low in cost, and have a large amount of screening samples. However, ELISA antibodies and enzymes are biologically active substances that require low-temperature storage and poor stability. ; Colloidal gold immunochromatography has low detection sensitivity and cannot perform accurate qualitative and quantitative determinations. It is mainly used for rapid screening analysis, and both enzyme-linked immunosorbent assay and colloidal gold immunochromatography have non-specific reactions, and the detection results are prone to appear false positive problem

Method used

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  • Fluorescence quantitative detection card for aflatoxin B1 and application thereof
  • Fluorescence quantitative detection card for aflatoxin B1 and application thereof
  • Fluorescence quantitative detection card for aflatoxin B1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: Preparation of Aspergillus flavus B1 fluorescence quantitative detection card

[0014] 1. Preparation of raw materials for Aspergillus flavus B1 fluorescence quantitative detection card

[0015] 1. Synthesis of Aspergillus flavus B1 artificial antigen

[0016] Aspergillus flavus B1 is a small molecule substance with only immunoreactivity, no immunogenicity, and cannot induce an immune response in the body. The two drugs are coupled to the carrier protein by the glutaraldehyde method or the diazotization method to obtain the two drugs The carrier protein conjugate is immunogenic.

[0017] 2. Preparation of Aspergillus flavus B1 monoclonal antibody

[0018] (1) Animal immunization program

[0019] BALB / c mice were used as immunized animals, Aspergillus flavus B1-carrier protein was used as immunogen, and the immunization dose was 50-100μg / mouse. In the first immunization, the immunogen and the same amount of Freund's complete adjuvant are mixed to make an emulsifier, an...

Embodiment 2

[0039] Example 2: Application of Aspergillus flavus B1 fluorescence quantitative detection card

[0040] 1. Sample preparation

[0041] 1. Blank sample: grain, feed

[0042] Grind the sample to be tested into powder (pass through a 20-mesh sieve); weigh 1.00 ± 0.05 g of the crushed sample into a 10 mL centrifuge tube; add 4 mL sample extract (sample extract: 70 mL methanol and 30 mL deionized Water, mix well), vortex vigorously for 5 min, 4000 g, and centrifuge for 5 min; take 50 μL of the processed sample supernatant and add it to 1 mL of sample diluent (0.01M PBS) and mix.

[0043] 2. Add sample: Add aflatoxin B1 standard to the blank sample to the required concentration.

[0044] 2. Determination of the lowest detection limit

[0045] Take 20 blank grain and feed samples, use the aflatoxin B1 fluorescence quantitative detection card for detection, calculate the average value, and add 3 times the standard deviation, which is the lowest detection limit.

[0046] Table 1 Statistical tabl...

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PUM

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Abstract

The invention discloses a fluorescence quantitative detection card for aflatoxin B1 and application thereof. The fluorescent quantitative detection card comprises a back plate, a sample pad, a quenching antibody binding pad, a fluorescent reaction film and an absorbent pad. According to the fluorescence quantitative detection card, the content of the aflatoxin B1 in sample matrixes like the cerealand fodder can be detected based on a principle of quenching the fluorescence obviously by using gold collected at a T area. The method is simple and is easy to implement; the preprocessing is simple; and the detection card has characteristics of high sensitivity, high specificity, high precision and high accuracy.

Description

Technical field [0001] The invention relates to a fluorescent quantitative detection card for detecting aflatoxin B1 in food and feed and its application. Background technique [0002] Aflatoxin B1 (Aflatoxin B1, AFB1) is a derivative of dihydrofuranoxaphthalone, which contains a difuran ring and an oxaphthalone (coumarin), which is highly carcinogenic and highly toxic. It is the most stable of all mycotoxins discovered so far. Since 1993, aflatoxin has been recognized as a carcinogen designated by the Cancer Research Institute of the World Health Organization (WHO), and it is one of the three most recognized carcinogens in the world. Aflatoxin B1 is the most toxic and polluting. It is the most harmful aflatoxin. Foods contaminated by aflatoxin B1 are mainly grain, oil, food and feed such as peanuts, corn, rice, wheat, and peanut oil. At present, 91 countries have established maximum residue limits (MRL) of TAFs in different foods. For example, the International Codex Alimenta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/532
CPCG01N33/558G01N33/532
Inventor 马立才刘河冰丁亚芳杨柳贾良曦覃丹凤秦誉
Owner 北京维德维康生物技术有限公司
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