Free triiodothyronine measurement kit and fabrication method thereof

A technology of triiodothyronine and protosine, applied in the field of in vitro testing, can solve the problems of drift of working curve over time, unfavorable automatic quantitative analysis, nonlinear downward shift of low-end slope, etc. Fast, well-performing results

Inactive Publication Date: 2019-01-18
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in ELISA and time-fluorescence resolution detection technology, the solid phase carrier is made of polystyrene small cup-shaped slats. The adsorption surface of this solid phase carrier is flat and smooth, and its reaction surface area and coated surface area Both are very limited, so the reaction time is long, the measurement speed is slow, and the linear range is narrow. When the concentration of the substance to be measured is high, the hook effect is prone to occur and wrong results occur. Since the coated solid-phase carrier is a reaction cup, the reaction cup is Determines the items to be measured, which is not conducive to automatic quantitative analysis like a pipeline
According to the labeling reaction system, it can be divided into two types: enzymatic and non-enzymatic. The detection method of enzymatic reaction is relatively simple, but there are deficiencies such as the drift of the working curve with time and the non-linear downward shift of the low-end slope.

Method used

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  • Free triiodothyronine measurement kit and fabrication method thereof
  • Free triiodothyronine measurement kit and fabrication method thereof
  • Free triiodothyronine measurement kit and fabrication method thereof

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Experimental program
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preparation example Construction

[0058] The preparation method of solid-phase reagent R1 is:

[0059] A. Activation of paramagnetic microspheres: take paramagnetic microspheres, wash with pH6.0~8.0, 10mM phosphate buffer, resuspend in the above buffer; use concentration of 0.5%~5% EDC solution to activate, in Under the condition of 30-40°C, shake and mix for 30-40 minutes to obtain paramagnetic microsphere resuspension;

[0060] B. Antigen coating: Add the T3 antigen analogue to the resuspension of paramagnetic microspheres, the feeding ratio is 0.2%-2%, and react at 25-37°C for 2-3 hours; then at 25-37°C Blocked for 20-40 minutes under certain conditions; wash the coated paramagnetic microspheres with pH 6.0-8.0, 10mM phosphate buffer, resuspend in the above buffer, and obtain T3 with a concentration of 0.05-0.5mg / ml Suspensions of analogue-coated paramagnetic microspheres;

[0061] The preparation method of liquid phase reagent R2 is:

[0062] Take the T3 antibody, dilute it with pH 9.5, 0.5M / L phosphate...

Embodiment 1

[0070] The preparation of embodiment 1 free triiodothyronine assay kit

[0071] Preparation of solid phase reagent R1:

[0072] A. Activation of paramagnetic microspheres: take the paramagnetic microspheres with -COOH groups on the surface, wash them twice with pH 6.0, 10mM phosphate buffer, remove the supernatant after magnetic separation; add the EDC prepared by the above phosphate buffer (concentration: 0.5%) solution, vortex and mix, and activate at 30°C for 40 minutes; remove the supernatant by magnetic separation, and then add the above-mentioned phosphate buffer solution to wash for 3 times to obtain a paramagnetic microsphere resuspension;

[0073] B. Antigen coating: Add the T3 antigen analogue to the resuspension of paramagnetic microspheres, the feeding ratio is 0.2%, react at 25°C for 3 hours, remove the supernatant by magnetic separation; then add a blocking agent such as containing TRIS buffer solution of protein or amino acid (concentration: 10%), block at 25°C...

Embodiment 2

[0076] The preparation of embodiment 2 free triiodothyronine determination kit

[0077] Preparation of solid phase reagent R1:

[0078] A. Activation of paramagnetic microspheres: take the paramagnetic microspheres with -COOH groups on the surface, wash them twice with pH 8.0, 10mM phosphate buffer, remove the supernatant after magnetic separation; add the EDC prepared by the above phosphate buffer (Concentration: 5%) solution, vortex and mix, activate at 40°C for 30 minutes; remove the supernatant by magnetic separation, then add the above-mentioned phosphate buffer to wash twice, and obtain a paramagnetic microsphere resuspension;

[0079] B. Antigen coating: Add the T3 antigen analogue to the resuspension of paramagnetic microspheres, the feeding ratio is 2%, react at 37°C for 2 hours, remove the supernatant by magnetic separation; then add a blocking agent such as containing TRIS buffer solution of protein or amino acid (concentration: 10%), block at 37°C for 20 minutes; ...

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Abstract

The invention discloses a free triiodothyronine measurement kit and a fabrication method thereof, and belongs to the technical field of vitro inspection. By the free triiodothyronine measurement kit,the technical problems of long reaction time, slow measurement speed, narrow of linear range and hook effect easy to generate of the prior art when a tested substance is high in concentration are solved. The kit comprises the following reagents of a solid-phase reagent 1 and a liquid-phase reagent, wherein the solid-phase reagent R1 is a suspension liquid including a T3 analogue-coated paramagnetism microsphere, the liquid-phase reagent R2 is a suspension liquid including an acridinium ester labeled T3 antibody, the paramagnetism microsphere in the R1 is Fe3O4 with a surface wrapped by a carboxyl active group, and the particle size of the paramagnetism microsphere is 0.1-5 micrometers. With the free triiodothyronine measurement kit and the fabrication method thereof, provided by the invention, free triiodothyronine in blood/plasma is quantitatively detected by a chemiluminescence immunity analysis method, the kit has the advantages of high sensitivity, rapid detection, wide linear range, low cost and the like and is simple and convenient to operate, and automation is facilitated.

Description

technical field [0001] The invention relates to the technical field of in vitro testing, in particular to a free triiodothyronine assay kit and a preparation method thereof. Background technique [0002] The thyroid gland is an endocrine gland, and the biologically active thyroid hormones it secretes have extensive and strong physiological effects on the body. It can promote growth and tissue differentiation, promote the oxidative decomposition of sugar, fat, and protein, increase oxygen consumption and heat production, increase the basic metabolic rate of the body, and play an important role in the central nervous system, cardiovascular and digestive systems. An increase or decrease in thyroid hormone secretion can lead to thyroid dysfunction and endocrine and metabolic disorders. Therefore, the correct detection of thyroid-related hormones is of great significance for the diagnosis and treatment of thyroid diseases. [0003] Triiodothyronine (3,5,3'-L-triiodothyronine, T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/78G01N33/543G01N33/553G01N33/577G01N33/58G01N33/532G01N21/76
Inventor 刘子菱刘婷婷何浩会王香琪阚洪晶丛凡淏付莉莉
Owner DIRUI MEDICAL TECH CO LTD
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