Colloidal gold immunochromatograohic assay (GICA) method of aflatoxin B1 in rice

A technology for immunochromatographic detection and aflatoxin, which is applied to measurement devices, analytical materials, instruments, etc., can solve problems such as inability to meet rapid on-site detection, cumbersome operations, etc., and achieve intuitive and reliable visual judgment, simple and fast operation, and high sensitivity. sexual effect

Inactive Publication Date: 2017-07-28
莆田方家铺子食品营养研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection method of above-mentioned prior art AFB1 needs special instrument and equipment and is cumbersome to operate, can't meet the requirement of on-the-spot rapid detection; Therefore, it has important practical significance to establish a kind of convenient, simple and fast AFB1 detection method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1 A kind of colloidal gold immunochromatographic detection method of aflatoxin B1 in rice

[0021] A colloidal gold immunochromatographic detection method for aflatoxin B1 in rice, comprising the following steps:

[0022] (1) Preparation and purification of anti-AFB1 monoclonal antibody: The hybridoma cell line 2A5 stably secreting anti-AFB1 monoclonal antibody was inoculated into the peritoneal cavity of BALB / c mice, 100 cells / mouse, 5 mice were inoculated, and obtained by in vivo induction 20ml anti-AFB1 ascites antibody, and then the ascites antibody was purified by 5ml affinity chromatography column to obtain 10mg high-purity anti-AFB1 monoclonal antibody;

[0023] (2) Preparation of colloidal gold immunochromatography detection system (GICA): take 0.01wt% HAuCl 4 Aqueous solution 100ml, heat and boil 30min, add 1wt% trisodium citrate aqueous solution 2.5ml rapidly, continue to boil about 5min, orange-red appears; Gained colloidal gold particle size is 2...

Embodiment 2

[0025] Embodiment 2 detects the positive rate of rice AFB1 of 30 different batches according to the detection method described in embodiment 1

[0026] Weigh 2.0g of crushed and homogenized rice sample, put it in a 50ml centrifuge tube, add 3ml of ethyl acetate, vortex and mix for 4min, centrifuge at 4000r / min for 4min, accurately measure 1ml of the supernatant, and pass through a 0.2um filter membrane. A rice sample solution was obtained.

[0027] Prepare 30 different batches of rice sample solutions according to the above method. The particle size of colloidal gold in the analytical probe is 20nm, slowly drop on the analytical probe, the pH value of the mark is 7.5, and the concentration of AFB1-BSA conjugate is 10 μg / mL, when goat anti-mouse IgG is the original solution, the detection limit of AFB1 is 10ng / mL by naked eyes, and the detection time is 10min; A red band appears, and the test sample is negative; when the AFB1 content in the sample exceeds 10ng / mL, the quality...

Embodiment 3

[0028] Embodiment 3 A colloidal gold immunochromatographic detection method of aflatoxin B1 in rice

[0029] A colloidal gold immunochromatographic detection method for aflatoxin B1 in rice, comprising the following steps:

[0030] (1) Preparation and purification of anti-AFB1 monoclonal antibody: The hybridoma cell line 2A5 stably secreting anti-AFB1 monoclonal antibody was inoculated in the peritoneal cavity of BALB / c mice, 110 cells / mouse, 4 mice were inoculated, and obtained by in vivo induction 25ml anti-AFB1 ascites antibody, and then the ascites antibody was purified by 5ml affinity chromatography column to obtain 12.5mg high-purity anti-AFB1 monoclonal antibody;

[0031] (2) Preparation of colloidal gold immunochromatography detection system (GICA): take 0.01wt% HAuCl 4 Aqueous solution 100ml, heat and boil 30min, add 0.75ml of 1wt% trisodium citrate aqueous solution rapidly, continue to boil about 5min, orange-red appears; Gained colloidal gold particle size is 50nm;...

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Abstract

The invention discloses a colloidal gold immunochromatograohic assay (GICA) method of aflatoxin B1 (AFB1) in rice. The method comprises the following steps: reducing HAuCl4 aqueous solution with a reduction method to prepare colloidal gold and uniformly mixing the colloidal gold and anti-AFB1 monoclonal antibody, so as to obtain a gold-labeled monoclonal antibody serving as an analysis probe; respectively enveloping an AFB1-BSA conjugate of a detection line and goat-anti-mouse IgG of a quality control line on a nitrocellulose membrane and establishing a GICA system of AFB1; slowing dropping rice sample solution onto the analysis probe and observing the color change of the detection line and the quality control line, wherein both the detection line and the quality control line are red when the concentration of AFB1 in a sample is lower than a limiting value; only the quality control line is red when the concentration of AFB1 in the sample is higher than the limiting value. The GICA technology disclosed by the invention serves as a novel detection technology, an instrument is not needed, and the technology has the advantages that the operation is simple and fast, and a detection result is timely and accurate. Therefore, the technology is suitable for field test for AFB1 and has wide application prospect in the food safety testing field.

Description

technical field [0001] The invention relates to the technical field of food detection, in particular to a colloidal gold immunochromatographic detection method for aflatoxin B1 in rice. Background technique [0002] Aflatoxin (AFs) is composed of difuran ring and oxinone, which is a strong carcinogen; aflatoxin B1 (AFB1) is the most toxic derivative of AFs, and the intake of AFB1 is the cause of One of the main factors of liver cancer. Feed and food crops with high moisture content and unsuitable storage temperature are easily contaminated by AFB1, and AFB1 can withstand high temperature, so it is difficult to be destroyed in the subsequent processing process, which seriously threatens the health of humans and animals. life safety. [0003] In order to reduce the harm of AFB1 to humans and animals, countries all over the world have continuously formulated the lowest possible limit standards for AFB1 in food and feed, and monitored them through inspection and quarantine agen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577
CPCG01N33/558G01N33/577
Inventor 方敏
Owner 莆田方家铺子食品营养研究中心
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