Multi-index combined-detection chemiluminiscence kit and preparation method and application thereof

A technology for chemiluminescence immunoassay and reagent analysis, which is applied in the field of multi-index joint detection chemiluminescence immunoassay kit and its preparation, and can solve the problems of high consumption of reagents, long time required for analysis, and low analysis throughput.

Inactive Publication Date: 2019-07-12
南京仁迈生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The analysis takes a long time, consumes a lot of reagents, the analysis throughput is low, and the labor load is large

Method used

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  • Multi-index combined-detection chemiluminiscence kit and preparation method and application thereof
  • Multi-index combined-detection chemiluminiscence kit and preparation method and application thereof
  • Multi-index combined-detection chemiluminiscence kit and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Simultaneous quantitative joint detection of multiple indicators of myocardial infarction (cTnT, Myo, CK-MB)

[0049] 1. Avidin-coated magnetic particles

[0050] 1. Take the stock solution of magnetic particles, take the reaction buffer solution to wash the magnetic particles, and then resuspend them in the buffer solution. Add a certain concentration of EDC ready-made solution to it, and activate it at 37°C for 0.5h. After activation, wash three times with magnetic particle washing solution, and then resuspend in reaction buffer solution.

[0051] 2. Add avidin and react at 37°C for 3h. After the reaction is completed, wash with magnetic particle cleaning solution three times, and finally resuspend in magnetic particle storage solution.

[0052] 2. Analysis steps

[0053] Samples 1-11 were prepared with PBS buffer solution; samples 1-11 contained cTnT: 1.0, 3.0, 9.0, 15.0, 40.0, 100.0, 500.0, 1000.0, 2000.0, 5000.0, 10000.0 ng / L, Myo: 15.0, 21.0, 42.0, 100.0, 200...

Embodiment 2

[0062] Simultaneous quantitative joint inspection of three indicators of thyroid function (TSH, FT3, FT4)

[0063] 1. Streptavidin-coated microplates

[0064] 1. Take a microwell plate, add coating solution containing streptavidin, and incubate at 37°C for 12h.

[0065] 2. After the incubation, wash three times on the plate washing machine with a special washing solution for coating, then add the blocking solution containing BSA, and incubate at 37°C for 30 minutes.

[0066] 3. After incubation, wash three times on the plate washing machine with special cleaning solution for coating, invert the microwell plate for 1 hour at a constant temperature of 25°C, drain the water in the microwell plate, and then transfer it to an airtight container at 25°C Aged for 36 hours in a constant temperature environment.

[0067] 2. Analysis steps

[0068] Prepare samples 1~11 with PBS buffer solution; samples 1~11 contain TSH: 0.002, 0.005, 0.01, 0.02, 0.05, 0.2, 1.0, 5.0, 20.0, 60.0, 100.0...

Embodiment 3

[0077] Simultaneous quantitative joint inspection of veterinary drug residue indicators (melamine, gentamicin, kanamycin)

[0078] 1. Magnetic particles coated with fluorescein isothiocyanate antibody

[0079] 1. Take the stock solution of magnetic particles, take the reaction buffer solution to wash the magnetic particles, and then resuspend them in the buffer solution. Add a certain concentration of EDC ready-made solution to it, and activate it at 37°C for 0.5h. After activation, wash three times with magnetic particle washing solution, and then resuspend in reaction buffer solution.

[0080] 2. Add fluorescein isothiocyanate antibody and react at 37°C for 3 hours. After the reaction is completed, wash with magnetic particle cleaning solution three times, and finally resuspend in magnetic particle storage solution.

[0081] 2. Analysis steps

[0082] Prepare samples 1-11 with PBS buffer solution; samples 1-11 contain melamine: 0.0025, 0.005, 0.01, 0.015, 0.03, 0.09, 0.2...

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Abstract

The invention discloses a multi-index combined-detection chemiluminiscence immunoassay method for forming quantum dot-enzyme complex based on quantum dot marker enzyme. The multi-index combined-detection chemiluminiscence immunoassay method for forming quantum dot-enzyme complex based on quantum dot marker enzyme comprises the following steps: marking quantum dots in different particle sizes or varieties on an enzyme surface or a polymer layer surface 5-10nm distancing from the enzyme, thereby forming different quantum dot-enzyme complex; the absorption spectrums of the quantum dots in different particles sizes or varieties and the enzyme are used for catalyzing the luminescent substrate to produce the emission spectrums, and the emission spectrums are sufficiently overlapped, and the quantum dots have large Strokes displacement, the emission light in 400-600nm wavelength produced by catalyzing the substrate by the enzyme can be efficiently converted into the emission light in the wavelength of 500-750nm, and the detection is performed under the effect of different optical filters; and the multi-index combined detection can be quickly and efficiently performed on a chemiluminescence platform by using less sample.

Description

technical field [0001] The invention belongs to the technical field of chemiluminescence immunity, and in particular relates to a multi-index joint detection chemiluminescence immunoassay kit based on a quantum dot-labeled enzyme forming a quantum dot-enzyme complex and its preparation method and application. Background technique [0002] Chemiluminescent immunoassay (Chemilμminescence immμnoassay, CLIA) is a combination of highly sensitive chemiluminescent assay technology and highly specific immune response, used for various antigens, haptens, antibodies, hormones, enzymes, fatty acids, vitamins and drugs and other detection and analysis techniques. It is a newest immunoassay technique developed after radioimmunoassay, enzyme immunoassay, fluorescent immunoassay and time-resolved fluorescent immunoassay. Compared with the traditional enzyme immunoassay, CLIA has the characteristics of higher sensitivity, shorter detection time, simpler labeling method and lower cost of ra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543
CPCG01N33/535G01N33/543G01N2446/00
Inventor 金晶王西龙张誉严
Owner 南京仁迈生物科技有限公司
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