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Sample loading method for time-resolved fluorescence immunoassay

A time-resolved fluorescence and immunoassay technology, applied in biological testing, material inspection products, etc., can solve the problems of prolonged operation time, experimental failure, increased error and error, etc., to increase the concentration of the marker, shorten the reaction time, and reduce the possibility of sexual effect

Active Publication Date: 2017-04-12
GUANGZHOU FENGHUA BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In this method of operation, since various reagents need to be prepared by the operator before each test, it is necessary to increase the errors caused by adding samples and the errors caused by different operators or different batches of operations.
On the other hand, since certain chemical reactions often occur between the various reagents and buffers involved in the reaction, or some reagents and buffers are prepared to the working concentration required by the instructions, which will affect the stability of the measurement results, so it cannot be Mix the reagents to be added and the buffer reagents before operation
At the same time, due to the introduction of an intermediate container, this method of adding samples is not only cumbersome to operate, but also prolongs the operation time, and increases the possibility of introducing contamination, which often leads to false positive or false negative results in the reaction, resulting in experimental failure.

Method used

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  • Sample loading method for time-resolved fluorescence immunoassay
  • Sample loading method for time-resolved fluorescence immunoassay
  • Sample loading method for time-resolved fluorescence immunoassay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The application of the sample adding method suitable for time-resolved fluorescent immunoassay in the determination of alpha-fetoprotein

[0025] 1) Prenatal screening alpha-fetoprotein (AFP) rapid assay kit

[0026] The main components in the alpha-fetoprotein rapid assay kit: (1) solid phase carrier; (2) lanthanide marker working solution, 4ml / bottle (directly added to the microwell plate); (3) buffer, 4ml / bottle (directly added to the microplate); (4) calibrator, 1.5ml / bottle; (5) concentrated washing solution and (6) enhancement solution;

[0027] The preparation method of solid-phase carrier: the AFP monoclonal antibody is coated buffer (can use 50mmol / L, the carbonic acid buffer of pH9.6, 20mmol / L, the phosphate buffer of pH4.5, 50mmol / L, pH7 .8 Tris-HCl buffer or 50mmol / L, pH4.5 citrate buffer) diluted to the optimum concentration, coated on the solid phase, washed the solid phase once, and then carried out with blocking solution Sealed, spin-dried the solid ...

Embodiment 2

[0100] The invention is applicable to the time-resolved fluorescent immunoassay adding method in the determination of neonatal thyroid stimulating hormone Applications

[0101] 1) Neonatal Thyroid Stimulating Hormone (TSH) Rapid Determination Kit

[0102] The main components in the Neonatal Thyroid Stimulating Hormone Rapid Determination Kit (96 servings / box, automatic comparison): (1) Solid phase carrier; (2) Lanthanide marker working solution, 4ml / bottle, 2 bottles (directly added to the microplate); (3) buffer solution, 4ml / bottle, 2 bottles (directly added to the microplate); (4) filter paper dried blood spot calibration product, 1 set; (5) concentrated washing solution and (6) enhancement fluid;

[0103] The preparation method of the solid phase carrier: the thyroid-stimulating hormone antibody is coated with a coating buffer (50mmol / L, carbonic acid buffer solution of pH9.6, 20mmol / L, phosphate buffer saline solution of pH4.5, 50mmol / L , pH7.8 Tris-HCl buffer or 50...

Embodiment 3

[0191] The application of the sample adding method suitable for time-resolved fluorescent immunoassay in the determination of rubella virus IgM

[0192] 1) Rubella virus IgM rapid assay kit

[0193] The main components in the rubella virus IgM rapid assay kit: 1) antibody solid phase carrier; 2) calibrator; 3) lanthanide markers, 4.0ml / bottle; 4) experimental buffer, 4.0ml / bottle; 5 ) Rubella virus IgM biotinylated antigen, 4ml / bottle 6) Concentrated lotion and 7) Enhancement solution.

[0194] The preparation method of solid-phase carrier: the anti-u chain is coated buffer (can use 50mmol / L, the carbonic acid buffer of pH9.6, 20mmol / L, the phosphate buffer of pH4.5, 50mmol / L, pH7. 8 Tris-HCl buffer or 50mmol / L, pH4.5 citrate buffer, etc.) to the optimum concentration, coat the solid phase, wash the solid phase once, and then use the blocking solution for Sealed, spin-dried the solid phase, air-dried, vacuum-packed, and stored at 2-8°C for later use.

[0195] The lanthani...

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Abstract

The invention discloses a sample adding method suitable for a time resolution fluorescence immunoassay method. The sample adding method comprises the steps of adding a sample into a solid phase carrier coated reaction plate, adding a buffer solution into each hole, and then adding a marker working solution, wherein the concentration of the marker working solution allows the marker working solution to be sufficiently mixed with to-be-measured materials in the sample when the volume of the buffer solution and that of the marker working solution both are not greater than 50 mu. L. According to the sample adding method suitable for the time resolution fluorescence immunoassay method, a step of mixing the buffer solution and a marker in an intermediate container is avoided, the operation is simple and convenient, the possibility of pollution introduction is reduced, errors about the same check and / or test among various laboratories, caused by different operators, even operations, in different batches, by the same operator are effectively reduced, the accuracy and credibility of test results are improved, and because the marker concentration in the marker working solution is increased, reaction time can be remarkably shortened.

Description

technical field [0001] The invention belongs to the technical field of time-resolved fluorescence immunoassay, in particular to a sample adding method suitable for time-resolved fluorescence immunoassay. Background technique [0002] When time-resolved fluorescent immunoassay is used to determine the content of substances in samples, the steps usually include: [0003] 1) Adding samples: mix the marker and buffer evenly in the container, and prepare the marker working solution; then add the sample to the solid-phase carrier-coated reaction plate, and add the prepared marker working solution to each well; [0004] 2) The reaction plate was incubated at room temperature with slow shaking; [0005] 3) Wash the plate with the above-mentioned working washing solution and pat dry; [0006] 4) Add enhancement solution to each well; shake the antibody solid phase carrier slowly at room temperature and then detect. [0007] Among them, the sampling method of the first step is very...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53
CPCG01N33/53
Inventor 陈建起鲍冬芹谭玉华
Owner GUANGZHOU FENGHUA BIOENG