Time-resolved fluoroimmunoassy test strip used for quantitative determination of soluble growth stimulation gene-2 (sST2), and preparation method thereof

A time-resolved fluorescence and quantitative detection technology, applied in the field of medical testing, can solve the problems that the soluble growth-stimulating gene 2 protein is not suitable for clinical rapid diagnosis, high requirements for testing equipment, and many interference factors, so as to facilitate the calculation of sample concentration and the detection time. The effect of short, accurate regression equations

Inactive Publication Date: 2017-05-31
天津欧尔克医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ELISA technology has the following disadvantages: high requirements for detection equipment, high cost; many interference factors, poor repeatability; long detection time
Therefore, ELISA detection of soluble growth-stimulating gene 2 protein is not suitable for rapid clinical diagnosis

Method used

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  • Time-resolved fluoroimmunoassy test strip used for quantitative determination of soluble growth stimulation gene-2 (sST2), and preparation method thereof
  • Time-resolved fluoroimmunoassy test strip used for quantitative determination of soluble growth stimulation gene-2 (sST2), and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The preparation method of the time-resolved fluorescent immunoassay test strip for quantitatively detecting sST2 comprises the following steps:

[0043] (1) Immobilizing sST2 monoclonal antibody II and rabbit anti-mouse IgG antibody recognizing different epitopes at different positions of the coating membrane 4 to form a detection line and a quality control line;

[0044] (2) prepare rare earth ion microsphere-labeled sST2 monoclonal antibody I, and spray on the binding pad 3;

[0045] (3) Paste the sample pad 2 , binding pad 3 , coating film 4 and absorbent paper 5 on the bottom plate 1 in sequence, and then cut them into 0.5 cm width and put them into the clamping case 8 .

[0046] The preparation method of coating film 4 in the step (1) is: use the phosphate buffer saline solution that the pH of 7.2 is 7.2 to use the 0.01mol / L that contains 1% sucrose, the sST2 monoclonal antibody II that recognizes different epitopes and the rabbit anti-mouse The IgG antibody was d...

Embodiment 2

[0049] The preparation method of the time-resolved fluorescent immunoassay test strip for quantitatively detecting sST2 comprises the following steps:

[0050] (1) Immobilizing sST2 monoclonal antibody II and rabbit anti-mouse IgG antibody recognizing different epitopes at different positions of the coating membrane 4 to form a detection line and a quality control line;

[0051] (2) prepare rare earth ion microsphere-labeled sST2 monoclonal antibody I, and spray on the binding pad 3;

[0052] (3) Paste the sample pad 2 , binding pad 3 , coating film 4 and absorbent paper 5 on the bottom plate 1 in sequence, and then cut them into 0.5 cm width and put them into the clamping case 8 .

[0053] The preparation method of coating film 4 in the step (1) is: use the phosphate buffer saline solution that the pH of 7.4 is 7.4 to use the 0.03mol / L containing 5.5% sucrose, respectively will recognize the sST2 monoclonal antibody II of different epitopes and the rabbit anti-mouse Dilute t...

Embodiment 3

[0056] The preparation method of the time-resolved fluorescent immunoassay test strip for quantitatively detecting sST2 comprises the following steps:

[0057] (1) Immobilizing sST2 monoclonal antibody II and rabbit anti-mouse IgG antibody recognizing different epitopes at different positions of the coating membrane 4 to form a detection line and a quality control line;

[0058] (2) prepare rare earth ion microsphere-labeled sST2 monoclonal antibody I, and spray on the binding pad 3;

[0059] (3) Paste the sample pad 2 , binding pad 3 , coating film 4 and absorbent paper 5 on the bottom plate 1 in sequence, and then cut them into 0.5 cm width and put them into the clamping case 8 .

[0060] The preparation method of coating film 4 in the step (1) is: use the phosphate buffer saline solution that the pH of 7.6 is 7.6 to use the 0.05mol / L containing 10% sucrose, the sST2 monoclonal antibody II that recognizes different epitopes and the rabbit anti-mouse The IgG antibody was dil...

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Abstract

The invention discloses a time-resolved fluoroimmunoassy test strip used for quantitative determination of sST2, and a preparation method thereof. The time-resolved fluoroimmunoassy test strip used for quantitative determination of sST2 comprises a base plate; a sample pad, a conjugate pad, a coating membrane, and an absorbent paper are arranged on the base plate in a staggered manner; the coating membrane is arranged on the middle part of the base plate; the upper end of one side edge of the coating membrane is covered by the conjugate pad, and the upper end of the other side edge is covered by the absorbent paper; and the edge of one side, far from the coating membrane, of the conjugated pad is covered by the sample pad. Compared with the prior art, the time-resolved fluoroimmunoassy test strip possesses following advantages: time-resolved fluoroimmunoassy is introduced into detection of soluble growth stimulation expressed gene 2 (sST2), quantitative determination of single portion sST2 is realized via combination with a time resolved fluorescence detector, sensitivity is high, intra-assay and inter-assay difference is low, great convenience is provided for clinical application, operation of the preparation method is simple, the preparation method is suitable for large scale production, and the preparation method possesses active significance in quantitative determination of sST2.

Description

technical field [0001] The invention belongs to the field of medical examination, and in particular relates to a time-resolved fluorescent immunoassay strip for quantitative detection of sST2 and a preparation method thereof. Background technique [0002] Growth-stimulating expression gene 2 protein (ST2) is a myocardial protein produced by cardiomyocytes under biomechanical stress. Tominaga et al. first discovered the ST2 gene in 1989, but it has been mistaken as an orphan because no functional ligand has been found. receptor. In 2005, Schmitz et al. discovered its specific functional ligand interleukin IL-33. The ST2 gene is expressed in mast cells, activated helper T cells (Th2), macrophages and cardiomyocytes. The human ST2 gene can encode a soluble growth-stimulating expression gene 2 protein (sST2) and a transmembrane growth-stimulating expression gene 2 protein (ST2L), and the transcription of the two is regulated by different promoters. [0003] IL-33 belongs to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/543G01N33/558G01N33/577G01N33/68
CPCG01N33/558G01N33/533G01N33/543G01N33/577G01N33/68
Inventor 陶剑周颖兰成杰
Owner 天津欧尔克医药科技有限公司
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