Fluorescent quantitative test strip capable of simultaneously detecting three mycotoxins as well as preparation method and application of fluorescent quantitative test strip

Abstract

The invention discloses a fluorescent quantitative test strip capable of simultaneously detecting mycotoxins (DON/ZEN/AFB1) as well as a preparation method and application of the fluorescent quantitative test strip. The preparation method comprises the following steps: firstly, marking specific antibodies and rabbit IgG of the three mycotoxins by using fluorescent microspheres, and then drying thespecific antibodies and the rabbit IgG on a fluorescent binding pad; sequentially marking detection lines T1, T2 and T3 of conjugates of the three mycotoxins and calf serum albumin and a quality control line C of goat anti-rabbit on the NC membrane; and finally, assembling a test strip, namely sequentially overlapping and adhering a sample pad, a fluorescent marker binding pad, a nitrocellulose membrane marked with three detection lines (T1/T2/T3 lines) and a quality control line (C line), and absorbent paper on PVC (polyvinyl chloride) cardboard, shearing into the test strip after assembling, and then loading into a plastic card shell. The test strip is high in sensitivity, small in intra-batch difference and inter-batch difference and good in repeatability, rapid quantitative detectionof three common mycotoxins in grains can be achieved within 8 min, and the result accuracy and operation simplicity of the test strip far exceed those of most immunochromatography rapid detection products on the market.

Description

technical field [0001] The invention belongs to the technical field of mycotoxin detection, and in particular relates to a fluorescent immunochromatographic test strip capable of simultaneously performing rapid and quantitative detection of mycotoxins (DON / ZEN / AFB1) in grains, and a preparation method and application thereof. Background technique [0002] Mycotoxin contamination in cereal feed is common and serious all over the world. All kinds of raw materials and feeds, all regions, and all seasons are contaminated with mycotoxins, and the detection rate can be as high as 70% to 100%, and the rate of toxins exceeding the standard determined according to feed hygiene standards can reach 20% to 80%. The toxins detected in the contaminated feed were as few as 2 to 3 types, and as many as 7 to 8 types, showing the characteristics of multiple mycotoxin compound pollution, and the structural formulas of several main toxins are as follows: [0003] [0004] According to the i...

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Problems solved by technology

Chromatography can be used for qualitative and quantitative analysis and determination of mycotoxins, and the results are reliable. However, the instruments used in chromatography are expensive and require professional personnel to operate. The pretreatment is complicated, the analysis period is long, and the cost is high. Generally, it is only suitable for Confirmatory testing

The protein phosphatase inhibition method is not yet mature enough. Many variables in the enzyme reaction system have not been quantified and unified standards have not been established, and the selectivity is poor. Many substances in the environment can interfere with the determination, and the results are not accurate enough. Therefore, practical applications are limited. very restrictive

Immunoassay is an analysis method based on antigen-antibody specific reaction, which has the characteristics of strong specificity, high sensitivity, fast detection speed, and low cost. It is currently a commonly used routine detection method, including enzyme-linked immunosorbent assay (ELISA) , colloidal gold rapid detection method, etc., but they all have their own advantages and disadvantages. Although the enzyme-linked immunosorbent assay has high detection sensitivity and accurate results, the operation is relatively complicated, the time is long, and only one toxin can be detected at a time; colloidal gold The method does not require pretreatment of the sample, and the detection is fast, but the sensitivity is relatively low, and only qualitative or semi-quantitative detection can be achieved, and quantitative detection results cannot be given.

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