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Fluorescence microsphere-colloidal gold double-coloring qualitative quantitative immunochromatographic test strip for detecting antibiotics and preparation method thereof

A technology of immunochromatographic test strips and fluorescent microspheres, which is applied in measurement devices, analytical materials, instruments, etc., can solve the problems of long time, large sample background interference, and narrow detection linear range, so as to avoid matrix interference and eliminate false positives. Positive results, the effect of saving testing time

Inactive Publication Date: 2018-11-30
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] ⑴General test strips are qualitatively analyzed by visual observation, and accurate quantitative detection cannot be achieved
[0012] (2) The matrix effect of different materials is obvious, and the sample background interference is large, which is easy to produce false positive results
[0013] (3) The positive result of the test cannot be saved, usually after the judgment time is exceeded, the result is no longer accurate and reliable
[0015] ⑴In the actual test, the proportion of negative results is very high, but all test strips of this method need to be read by an instrument, otherwise the results cannot be obtained, and it takes a long time to test a large number of samples!
[0016] (2) In quantitative detection, the detection department still needs a threshold to distinguish negative and positive, so the amount of data processing is relatively large
[0017] (3) The sensitivity of a single fluorescent microsphere test strip is high, but in practical applications, the detection linear range of the higher sensitivity is narrower, and the quantitative concentration range of the sample is not suitable for the actual detection requirements

Method used

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  • Fluorescence microsphere-colloidal gold double-coloring qualitative quantitative immunochromatographic test strip for detecting antibiotics and preparation method thereof
  • Fluorescence microsphere-colloidal gold double-coloring qualitative quantitative immunochromatographic test strip for detecting antibiotics and preparation method thereof
  • Fluorescence microsphere-colloidal gold double-coloring qualitative quantitative immunochromatographic test strip for detecting antibiotics and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Example 1: Preparation of fluorescent microspheres-colloidal gold dual-color qualitative and quantitative immunochromatographic test strips for detection of chloramphenicol (optimum ratio of fluorescent microspheres and colloidal gold antibody-labeled complexes)

[0096] 1. Preparation process of immunochromatographic test strips

[0097] 1. Preparation of Nitrocellulose Membrane

[0098] (1) Preparation of chloramphenicol artificial antigen (CAP-BSA)

[0099] The coupling method is the active lipid method, and the coupling ratio is 1:10-1:100. After coupling, dialysis purification is used to obtain CAP-BSA.

[0100] (2) Preparation of detection line and quality control line

[0101] The CAP-BSA conjugate and anti-mouse antibody were coated on the nitrocellulose membrane: Dilute the CAP-BSA conjugate with 0.01M pH 7.5 PBS (phosphate buffer saline) to a concentration of 0.6 mg / mL, and the resulting solution Spray on the membrane as a test line; the concentration of th...

Embodiment 2

[0119] Example 2: Preparation of fluorescent microsphere-colloidal gold dual-color qualitative and quantitative immunochromatographic test strips for detection of chloramphenicol (the proportion of fluorescent microsphere-labeled complexes increases)

[0120] The difference from Example 1 is:

[0121] ⑴After the preparation of the fluorescent microsphere antibody complex and the colloidal gold antibody complex is completed, calculate according to the amount of antibody on different markers in each test strip. The concentration of the antibody is 10ng / strip to spray the conjugated pad, spray it on a 30×0.8cm glass fiber membrane, dry it in vacuum at 25°C for 1-2 hours, and put it in a drying cabinet for later use. The test result is: the concentration of the standard curve under this condition is: 0, 0.01, 0.03, 0.05, 0.1, 0.3ng / mL, R 2 0.9176, IC 50 0.17ng / mL, poor linearity. Colloidal gold visual inspection was normal.

[0122] (2) Assemble the test strip: the test is the...

Embodiment 3

[0125] Example 3: Preparation of fluorescent microspheres-colloidal gold dual-color qualitative and quantitative immunochromatographic test strips for detection of chloramphenicol (the proportion of colloidal gold-labeled complexes increases)

[0126] The difference from Example 1 is:

[0127] (1) After the fluorescent microsphere antibody complex and the colloidal gold antibody complex are prepared, calculate according to the amount of antibody on different markers in each test strip. The concentration of the antibody was 6ng / strip to spray the conjugated pad, spray it on the glass fiber membrane of 30×0.8cm, dry it in vacuum at 25°C for 1-2h, and put it in the drying cabinet for later use. The test result is: the concentration of the standard curve under this condition is: 0, 0.01, 0.03, 0.05, 0.1, 0.3ng / mL, R 2 0.9287, IC 50 0.12ng / mL, poor linearity. The sensitivity of colloidal gold decreased to about 0.3ng / mL.

[0128] (2) Assemble the test strip: the test is the sam...

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Abstract

The invention discloses a fluorescence microsphere-colloidal gold double-coloring qualitative quantitative immunochromatographic test strip for detecting antibiotics and a preparation method thereof.A bottom plate is successively overlapped and pasted with a filter paper, a sample pad, a glass fiber pad coated with a fluorescence microsphere-antibiotic monoclonal antibody compound and a colloidalgold-antibiotic monoclonal antibody compound, a nitrocellulose membrane and an absorbent paper; the nitrocellulose membrane is coated with an antibiotic conjugated antigen as a detection line and coated with an anti-mouse antibody as a quality control line; if the color development of the detection line is lighter than that of the quality control line, then the qualitative judgment of the antibiotics in the samples is positive; and the positive test strip is directly inserted into a fluorescence reader which quantizes fluorescence signals of a fluorescence microsphere color system to realizequantitative detection. The Fluorescence microsphere-colloidal gold double-coloring qualitative quantitative immunochromatographic test strip for detecting the antibiotics is mainly used for qualitative and quantitative detections of the antibiotics in food safety detection.

Description

technical field [0001] The invention belongs to the field of antibiotic residue detection in food safety, and specifically relates to a time-resolved fluorescent microsphere-colloidal gold dual-color qualitative and quantitative immunochromatographic test strip capable of qualitatively and quantitatively detecting antibiotics in a sample and a preparation method thereof. Background technique [0002] Antibiotics are a class of secondary metabolites with anti-pathogen or other activities produced by microorganisms or higher animals and plants during their life, and chemical substances that can interfere with the development and function of other living cells. It is mainly used to treat various bacterial infections or pathogenic microbial infections, but the abuse of antibiotics can lead to liver and kidney damage, flora imbalance, antibiotic-associated diarrhea, and increased bacterial resistance, superinfection, and even delays Severe adverse reactions such as death. [000...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N33/543G01N33/558
CPCG01N33/533G01N33/54313G01N33/558
Inventor 赖卫华陈媛彭娟章刚钢黄震邢克宇熊勇华魏华许恒毅
Owner NANCHANG UNIV
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