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High-sensitivity immunochromatographic test strip for detecting total aflatoxin content quickly and preparation method thereof

An immunochromatographic test paper and aflatoxin technology, which is applied in the direction of measuring devices, biological testing, and analytical materials, can solve the problems of not being able to meet the testing requirements, interference of testing results, cross-contamination, etc. Effects with high application value and simple operation

Active Publication Date: 2010-12-29
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are rapid detection test strips for aflatoxin single component such as aflatoxin B1 (AFB1), but because most samples contaminated by aflatoxin contain more than one aflatoxin AFB1, it is easy to Cause interference or cross-contamination to the test results, and the sensitivity of commercially available aflatoxin immunochromatographic test strips is relatively low, with a detection limit of 5-200ng / g, which cannot well meet the current stricter aflatoxin limits Standard Testing Requirements

Method used

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  • High-sensitivity immunochromatographic test strip for detecting total aflatoxin content quickly and preparation method thereof
  • High-sensitivity immunochromatographic test strip for detecting total aflatoxin content quickly and preparation method thereof
  • High-sensitivity immunochromatographic test strip for detecting total aflatoxin content quickly and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: the preparation method of anti-aflatoxin universal monoclonal antibody

[0049] The anti-aflatoxin universal monoclonal antibody is produced by the hybridoma cell line 1C11 whose preservation number is CCTCC NO.C201013, and its preparation method comprises the following steps:

[0050] 1. Animal immunization

[0051] Six 6-week-old BALB / c mice were purchased and immunized with commercially available aflatoxin complete antigen AFB1-BSA. For the first immunization, the aflatoxin complete antigen was emulsified with an equal amount of Freund's complete adjuvant, and then injected subcutaneously at multiple points on the back of the neck of the mouse. The second immunization was carried out 4 weeks later, and the complete antigen was emulsified with the same amount of aflatoxin in Freund's incomplete adjuvant, and injected intraperitoneally into the mice. The interval between the third immunization and the second immunization was 4 weeks, and the immunizatio...

Embodiment 2-4

[0063] Example 2-4: Preparation and application of high-sensitivity immunochromatographic test strips for rapid detection of the total amount of aflatoxin

[0064] The anti-aflatoxin universal monoclonal antibody used in the following Examples 2-4 is the anti-aflatoxin universal monoclonal antibody prepared in Example 1.

Embodiment 2

[0066]A method for preparing a high-sensitivity immunochromatographic test strip for rapid detection of the total amount of aflatoxin, comprising the following steps:

[0067] (1) Preparation of absorbent pad

[0068] Cut the absorbent paper to a size of 16mm and 3mm wide to obtain an absorbent pad;

[0069] (2 Preparation of detection pad

[0070] The coating of the detection line:

[0071] Prepare aflatoxin B1-bovine serum albumin conjugate AFB1-BSA into 0.1mg / mL coating solution A; spray coating solution A at a position 15mm away from the upper edge of the nitrocellulose membrane Horizontally coated on a nitrocellulose membrane to obtain a detection line, the coating amount of aflatoxin B1-bovine serum albumin conjugate required per centimeter of detection line is 80ng, and then dried at 37°C for 15 minutes;

[0072] The coating liquid A is: 10 mg commercially available aflatoxin B1-bovine serum albumin conjugate AFB1-BSA, 1 g bovine serum albumin, 1 g sucrose, 0.02 g so...

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Abstract

The invention belongs to the field of biological detection and provides a high-sensitivity immunochromatographic test strip for detecting total aflatoxin content quickly, which is characterized by comprising a test strip, wherein a water-absorbing pad, a detection pad, a gold label pad and a sample pad are adhered on one side of the test strip from top down; the pads are overlapped at the connected parts; the detection pad uses a nitrocellulose film as a substrate pad; transverse quality control lines and detection lines are arranged on the nitrocellulose film from top down; the detection lines are coated with aflatoxin B1-bovine serum albumin (AFB1-BSA) coupling and the quality control lines are coated with rabbit anti-mouse polyclonal antibody; and a nano gold-labeled anti-aflatoxin common monoclonal antibody, which is generated by hybrid tumor cell strain 1C11 having a collection number of CCTCC No.C201013, is sprayed on the gold label pad transversely. The test strip is used for detecting total aflatoxin content and has the characteristics of quick detection, simple operation and high flexibility.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a high-sensitivity immunochromatographic test strip for rapidly detecting the total amount of aflatoxin and a preparation method thereof. Background technique [0002] Aflatoxin is mainly a secondary metabolite secreted by Aspergillus flavus and Aspergillus parasiticus, and is a natural toxic compound that can cause various damages to humans and animals. More than 20 kinds of aflatoxin (AFT) have been found, the most important of which are aflatoxin B1, B2, G1 and G2. [0003] After aflatoxin contaminates food and feed, it will directly or indirectly enter the human food chain, threatening human health and life safety, and the degree of harm is proportional to the intake of aflatoxin. Aflatoxins widely exist in rice, corn, peanuts, sesame, soybeans, rapeseed and other agricultural products and fish and other foods. There are many links to contaminate food and feed...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531G01N33/52G01N33/558
Inventor 李培武张道宏张奇张文丁小霞姜俊陈小媚
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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