High-flux ultra-sensitive double-label time-resolved immunochromatographic test strip and application thereof

A time-resolved fluorescence, test strip technology, applied in analytical materials, biological material analysis, measurement devices, etc., can solve the problem that the sensitivity is difficult to meet people's requirements, and achieve the effects of rapid detection, high sensitivity and good stability

Inactive Publication Date: 2020-03-10
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sensitivity of traditional colloidal gold ma

Method used

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  • High-flux ultra-sensitive double-label time-resolved immunochromatographic test strip and application thereof
  • High-flux ultra-sensitive double-label time-resolved immunochromatographic test strip and application thereof
  • High-flux ultra-sensitive double-label time-resolved immunochromatographic test strip and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1: Preparation of aflatoxin B1 and zearalenone double-labeled time-resolved immunochromatographic test strips

[0076] 1. Preparation of time-resolved fluorescent microsphere-conjugated antibody probes

[0077] 1. Time-resolved fluorescent microspheres conjugated to aflatoxin B1 monoclonal antibody

[0078] (1) Add 5 μL of time-resolved fluorescent microsphere solution to 500 μL of 0.2 mol / L borate borax buffer (pH 8.0), vortex and mix well, add 1 μg EDC and 1 μg NHS respectively, and activate on a shaker in the dark 15min.

[0079] (2) After completing step (1), add 8 μg of aflatoxin B1 monoclonal antibody (purchased from Beijing Weideweikang Biotechnology Co., Ltd., catalog number AFB-12A8), and react at room temperature for 2 hours or overnight at 4°C.

[0080] (3) After completing step (2), add 50 μL of 10% BSA solution containing 1% Tween 20, and block at room temperature on a shaker for 2 hours to fully block redundant activation sites on the surface of ...

Embodiment 2

[0100] Example 2: Qualitative and quantitative detection of aflatoxin B1 and zearalenone

[0101] 1. Qualitative detection method of aflatoxin B1 and zearalenone (line elimination method)

[0102] 1. Take 1 g of sample and add 4 mL of 70% (volume fraction) methanol solution. After shaking for 5 minutes, centrifuge at 8000 rpm for 10 minutes. Collect the supernatant and dilute it 10 times with sample diluent to obtain the processed sample. Take 120 μL of the treated sample, add 1 μL of the time-resolved fluorescent microsphere-coupled aflatoxin B1 monoclonal antibody solution prepared in Example 1, and 1 μL of the time-resolved fluorescent microsphere-coupled zearalen prepared in Example 1 The ketone monoclonal antibody solution and 1 μL of the time-resolved fluorescent microsphere-coupled chicken IgY antibody solution prepared in Example 1 were mixed evenly to obtain a mixed solution.

[0103] 2. After completing step 1, incubate the mixed solution for 5 minutes, then add the...

Embodiment 3

[0116] Example 3: Application of double-labeled time-resolved immunochromatographic method

[0117] 1. Preparation of samples to be tested

[0118] Coix seed, Chinese yam, and Campanulaceae that were identified by high-performance liquid chromatography-secondary mass spectrometry (UPLC-MS / MS) as samples that do not contain aflatoxin B1 and zearalenone were processed as follows: the samples were Grind to obtain a ground sample; take 1 g of the ground sample and add 4 mL of 70% (volume fraction) methanol solution, shake for 5 minutes, then centrifuge at 8000 rpm for 10 minutes, collect the supernatant and dilute it 10 times with the sample diluent to obtain the processed sample .

[0119] Add different concentrations of aflatoxin B1 and zearalenone to the treated samples to obtain test sample solutions containing different concentrations of aflatoxin B1 and zearalenone. Taking coix seed as an example, when the addition amount of AFB1 in the sample solution to be tested is 1 μg...

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Abstract

The invention discloses a high-flux ultra-sensitive double-labeled time-resolved immunochromatographic test strip and application thereof. The high-flux ultra-sensitive double-label time-resolved immunochromatographic test strip comprises a sample pad, a nitrocellulose film, a water absorption pad and a PVC bottom plate, wherein two detection lines and a quality control line are arranged on the nitrocellulose film; the detection line is respectively coated with antigens of a small molecular compound A and a small molecular compound B; and the quality control line is coated with a goat anti-chicken antibody. The invention also discloses a complete set of time-resolved fluorescent microsphere coupled antibody probe, which consists of a time-resolved fluorescent microsphere labeled anti-smallmolecule compound A antibody, a time-resolved fluorescent microsphere labeled anti-small molecule compound B antibody and a time-resolved fluorescent microsphere labeled chicken IgY antibody. The test strip and the complete set of time-resolved fluorescent microsphere coupled antibody probe can be used for rapidly, sensitively and accurately detecting various small molecular harmful compounds.

Description

technical field [0001] The invention belongs to the detection of small molecule harmful compounds in the field of food safety, and in particular relates to a high-throughput ultrasensitive double-labeled time-resolved immunochromatographic test strip and an application thereof. Background technique [0002] Immunochromatography technology is an immunoassay technology that has rapidly emerged in recent years by integrating the advantages of immunology technology and chromatography technology. This method has been widely used in the detection of small molecules. The nitrocellulose membrane material is used as the solid phase, and the sample to be tested is used as the mobile phase. Through capillary chromatography, the analyte reacts specifically with the receptors on the membrane for the analyte. The analyte reacts with the detection line on the NC membrane, and the detection result is determined by detecting or visually observing the color of the marker. In addition to the...

Claims

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Application Information

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IPC IPC(8): G01N33/58G01N33/543G01N33/531
CPCG01N33/531G01N33/54313G01N33/582G01N33/585G01N2430/00
Inventor 江海洋郑丕苗王战辉沈建忠温凯吴聪明史为民曹兴元丁双阳李建成张艳芳王梓乐
Owner CHINA AGRI UNIV
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