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A quantum dot immunochromatographic test strip for rapid detection of Brucella antibodies

An immunochromatographic test strip, Brucella technology, applied in measurement devices, biological tests, analytical materials, etc., can solve the problems of low sensitivity, low specificity, unsuitable for rapid detection, cumbersome operation, etc. Simple and fast labeling process, improving the effect of experimental techniques

Active Publication Date: 2021-08-31
ICDC CHINA CDC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, SAT has low sensitivity and specificity, and the operation is cumbersome and time-consuming, so it is not suitable for rapid detection of on-site epidemics.

Method used

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  • A quantum dot immunochromatographic test strip for rapid detection of Brucella antibodies
  • A quantum dot immunochromatographic test strip for rapid detection of Brucella antibodies
  • A quantum dot immunochromatographic test strip for rapid detection of Brucella antibodies

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] The preparation process of embodiment 1 Brucella whole bacterial protein

[0059] Select Brucella 16M with good antigenicity to prepare whole bacterial protein.

[0060] 1. Collect bacteria Take out the well-growth culture on the culture medium, heat and sterilize it in a water bath at 70-80°C for 1 hour, centrifuge, discard the supernatant, and collect the precipitated bacteria.

[0061] 2. Collect whole bacterial protein Suspend the thalline collected in the above step 1 in 0.5% carbolic acid saline, so that the concentration of the suspension is more than 2 times that of the agglutination antigen stock solution of the Brucella test tube. Then put the suspension at 108°C under steam pressure and heat it for 40-60 minutes, and put the heated bacteria suspension in a cool and dark place for more than two weeks. The supernatant is extracted by centrifugal precipitation, and the supernatant is sterile filtered, and the filtrate is the whole bacteria protein of Brucella. ...

Embodiment 2

[0062] Example 2 Detecting the preparation method of Brucella antibody quantum dot immunochromatography test strip

[0063] 1. Quantum dot activation and coupling process:

[0064] a. 25ul carboxylated CdSe / ZnS core-shell quantum dot QDs 610nm Solution, 5ul 0.1mol / LpH6.0MES solution and 20ul ultrapure water were added to a 1.5ml centrifuge tube, mixed briefly and centrifuged for 10s;

[0065] b. Weigh the activators EDC and NHS, prepare them to 1.9mg / ml and 2.1mg / ml respectively, add 5ul and 7.5ul respectively into the centrifuge tube of step a, mix quickly and place in a water-proof thermostat at 37°C The incubator reacted for 15 minutes;

[0066]c. After the reaction is completed, ultrasonically disperse it for 2 minutes, and then place the above-mentioned reaction product in a low-temperature high-speed centrifuge for centrifugation. The centrifugal force is set to 8000 rcf, and the centrifugation time is 20 minutes. After centrifugation, discard the supernatant, add 25u...

Embodiment 3

[0083] Example 3 Detection of Brucella Antibody Quantum Dot Immunochromatography Test Strip Preparation Method Optimization Experiment

[0084] 1. Optimization of reaction conditions for quantum dot-labeled Brucella whole bacterial protein:

[0085] 1.1 Optimization and determination of the optimal dosage of Brucella whole bacterial protein

[0086] Set the amount of antigen for labeling to 10ug, 15ug, and 20ug respectively. After labeling, use an immunofluorescence analyzer to detect the fluorescence intensity of the T-line, and observe the fluorescence intensity of the detection line after three different amounts of the quantum dot-labeled antigen. Finally, 15ug was used as the optimal amount of conjugated antigen, and the results are shown in image 3 .

[0087] 1.2 Optimization and determination of the ratio of activator to quantum dots

[0088] Take 25ul of quantum dots as the basic dosage, and set the volume ratio of EDC:NHS:quantum dots to 1:1.5:5, 1:2:5, 1.5:1:5, 2:...

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Abstract

The invention provides a quantum dot immunochromatographic test strip for rapid detection of Brucella antibodies, which is prepared by pasting a sample pad, a binding pad, a chromatographic membrane, and a water-absorbing pad on a PVC base plate in sequence by 1-1.5mm overlapping each other , the chromatographic membrane is a solid-phase nitrocellulose membrane composed of a detection line and a quality control line, the detection line is coated with Brucella whole bacteria protein, and the quality control line is coated with a secondary antibody, The binding pad is coated with quantum dot-labeled Brucella whole bacterial protein, and the present invention also provides a method for preparing the test strip, which greatly simplifies the labeling steps and improves the labeling efficiency; the prepared test strip shortens the The detection time is long, and it also has the advantages of strong specificity, good stability, and less serum consumption. It can realize the detection of Brucella antibodies in human and animal serum, and is of great importance for the rapid on-site screening and diagnosis of Brucellosis. significance.

Description

technical field [0001] The invention relates to a quantum dot immunochromatographic test strip, in particular to a quantum dot immunochromatographic test strip for detecting Brucella antibodies. Background technique [0002] Brucellosis is a widespread and serious bacterial infectious disease. my country's "Law on the Prevention and Control of Infectious Diseases" lists brucellosis as a Class B infectious disease. The disease is a zoonotic infectious disease caused by Brucella, with about 500,000 new cases each year. The human body is mainly caused by contact with infected animals or consumption of unpasteurized milk products. At present, there are 12 kinds of pathogenic factors of Brucella isolated, namely: sheep (B.melitensis), cattle (B.abortus), pigs (B.suis), sheep epididymis (B.ovis) , B.canis, B.neotomae, B.ceti, B.pinnipedialis, B.neotomae, Brucella vole ( B.microti), isolated from the human body (B.inopinata), isolated from the baboon species (B.papionis) and isola...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558G01N33/68G01N33/569G01N33/533
CPCG01N33/533G01N33/558G01N33/56911G01N33/6854G01N2333/23G01N2333/47
Inventor 姜海李广强王升启荣振赵鸿雁
Owner ICDC CHINA CDC
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