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Aflatoxin B1 flow lag immunization time distinguishing fluorescence rapid-detection kit and application thereof

A time-resolved fluorescence and aflatoxin technology, applied in the direction of fluorescence/phosphorescence, measuring devices, analytical materials, etc., can solve the problems of expensive instruments, cumbersome operation, interference of other components, etc., and achieve high accuracy and simple operation

Active Publication Date: 2013-09-04
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, thin-layer chromatography is the most commonly used detection method for the detection of aflatoxin earlier. This method does not require special equipment and can be carried out in general laboratories, but the amount of reagents is large, the operation is cumbersome, and other components interfere seriously. , poor accuracy, cannot be accurately quantified, and is more harmful to the experimenters and the surrounding environment, so it is not suitable for on-site rapid detection
Precision instrument analysis mainly includes fluorescence spectrophotometry and high-performance liquid chromatography. These methods have high sensitivity and good accuracy, but they are expensive and require a high degree of purification of aflatoxin samples. The sample pretreatment process is cumbersome and time-consuming. High requirements for the experimental environment, it is difficult to achieve rapid detection

Method used

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  • Aflatoxin B1 flow lag immunization time distinguishing fluorescence rapid-detection kit and application thereof
  • Aflatoxin B1 flow lag immunization time distinguishing fluorescence rapid-detection kit and application thereof
  • Aflatoxin B1 flow lag immunization time distinguishing fluorescence rapid-detection kit and application thereof

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Experimental program
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Embodiment 1

[0031] The acquisition of embodiment 1 anti-aflatoxin B1 monoclonal antibody

[0032] The anti-aflatoxin B1 monoclonal antibody is a monoclonal antibody produced by the secretion of the hybridoma cell line AFB3G1 with the preservation number CCTCC NO.C201015. The specific preparation method:

[0033]The hybridoma cell line AFB3G1 was injected into BALB / c mice treated with Freund's incomplete adjuvant in advance, the ascites of the mice was collected, and the antibody was purified by octanoic acid-ammonium sulfate method. Rat ascites, 4°C, centrifuged at 12000r / min for 15min, sucked the supernatant, mixed the obtained ascites supernatant with 4 times the volume of acetate buffer, slowly added n-octanoic acid under stirring, the volume of n-octanoic acid required per milliliter of ascites was 33μL, mix at room temperature for 30min, stand at 4°C for 2h, then centrifuge at 12000r / min for 30min at 4°C, discard the precipitate, filter the obtained supernatant with double-layer filt...

Embodiment 4

[0051] Example 4: Aflatoxin B1 Flow Hysteresis Immunization Time-Resolved Fluorescence Rapid Test Kit and Its Application

[0052] Aflatoxin B1 flow lag immune time-resolved fluorescence rapid test kit, which includes fluorescent test strips, sample reaction bottles containing europium-labeled anti-aflatoxin B1 monoclonal antibody freeze-dried products, sample diluent and sample diluent pipette , the fluorescent test strip includes cardboard, and one side of the cardboard is pasted with water-absorbing pads, detection pads and sample pads sequentially, and adjacent pads are overlapped and connected at the joints, and the overlapping length is 1mm, wherein: water-absorbing pads Length 12mm, width 3mm; detection pad length 25mm, width 3mm; sample pad length 15mm, width 3mm. The detection pad is based on a nitrocellulose membrane, and a horizontal quality control line and a detection line are arranged on the nitrocellulose membrane from top to bottom, and the quality control line...

Embodiment 5

[0083] Example 5: Aflatoxin B1 Flow Hysteresis Immunization Time-Resolved Fluorescence Rapid Test Kit and Its Application

[0084] Aflatoxin B1 flow lag immune time-resolved fluorescence rapid test kit, which includes fluorescent test strips, sample reaction bottles containing europium-labeled anti-aflatoxin B1 monoclonal antibody freeze-dried products, sample diluent and sample diluent pipette , the fluorescent test strip includes cardboard, one side of the cardboard is pasted with water-absorbing pads, detection pads and sample pads in sequence, and adjacent pads are overlapped and connected at the joints, and the overlapping length is 2mm, wherein: water-absorbing pads Length 15mm, width 3mm; detection pad length 28mm, width 3mm; sample pad length 12mm, width 3mm. The detection pad is based on a nitrocellulose membrane, and a horizontal quality control line and a detection line are arranged on the nitrocellulose membrane from top to bottom, and the quality control line is c...

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Abstract

The invention relates to an aflatoxin B1 flow lag immunization time distinguishing fluorescence rapid-detection kit and an application thereof. The kit comprises a fluorescent test strip and a sample reaction bottle containing an europium-labeled anti-aflatoxin B1 monoclonal antibody lyophilized product, wherein the fluorescent test strip comprises a cardboard, a water absorption pad, a detection pad and a sample pad are sequentially pasted on one surface of the cardboard from top to bottom, adjacent pads are connected at the connection in an overlapping manner, the detection pad treats a cellulose nitrate membrane as a base pad, a transverse quality control line and a detection line are arranged on the cellulose nitrate membrane from top to bottom, the quality control line is coated with a rabbit anti-mouse polyclonal antibody, and the detection line is coated with an aflatoxin B1 bovine serum albumin conjugate; and the anti-aflatoxin B1 monoclonal antibody is secreted by a hybridoma cell strain having a preservation number of CCTCC NO.C201015. The kit can be used for the quantitative determination of the content of the aflatoxin B1, and has the advantages of simple operation, rapidness and high accuracy.

Description

technical field [0001] The invention relates to aflatoxin B1 flow hysteresis immune time-resolved fluorescence rapid test kit and application thereof. Background technique [0002] Aflatoxins are mainly secondary metabolites secreted by Aspergillus flavus and Aspergillus parasiticus, and are natural toxic compounds that can cause various damages to humans and animals. More than 20 types of aflatoxins have been discovered, mainly including aflatoxin B1 (AFB1), B2 (AFB2), AFG and M1 (AFM1). Among the mycotoxins that have been discovered so far, aflatoxin B1 (AFB1) is the most toxic, and its toxicity, carcinogenicity and pollution frequency all rank first in biological toxins. Aflatoxin has many possible pollution links in food and feed. It is widely found in rice, corn, peanut, sesame, soybean, rapeseed and other agricultural products and fish. After AFB1 contaminates food and feed, it will directly or indirectly Entering the human food chain, threatening human health and li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N21/64
Inventor 李培武李冉张奇丁小霞张文张兆威
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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