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A soluble ST2 enzyme-linked immunoassay kit and preparation method thereof

An enzyme-linked immunoassay detection and detection kit technology, which is applied in the field of immunoassay medicine, can solve the problems of less disadvantageous clinical sample detection, cumbersome process, increase detection cost, etc., so as to reduce detection cost and labor cost, and simplify operation steps. , the effect of shortening the detection time

Active Publication Date: 2019-01-18
TIANJIN KANGERKE BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The detection time of this product is long, and the experimental samples need to be diluted in two steps in advance; the built-in high-concentration calibrator needs to be re-thawed in advance and diluted to each standard point during the detection, the process is relatively cumbersome and easy to introduce operational errors; streptavidin The vegetarian-horseradish peroxidase conjugate also needs to be diluted
The kit is not equipped with quality control products, and experimental operators need to purchase additional calibration materials when performing quality control on the detection system, which increases the cost of detection
In addition, the product only provides 96 servings of packaging, and the calibrator is only valid for 7 days after reconstitution, which is not conducive to domestic small and medium-sized medical institutions to test the small number of clinical samples that may be faced in the initial stage of the project

Method used

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  • A soluble ST2 enzyme-linked immunoassay kit and preparation method thereof

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Effect test

Embodiment 1

[0039] A preparation method of a soluble ST2 enzyme-linked immunoassay kit, comprising the following steps: 1) Preparation of soluble ST2 standard and quality control products: take an appropriate amount of finely ground ammonium sulfate and add it while stirring at room temperature (18-25° C.) Fetal bovine serum to a final concentration of 100g / L, placed at 4°C for 18 hours; centrifuged at 10,000 rpm for 20 minutes, and filtered the supernatant with filter paper; put the filtered supernatant into a dialysis bag, and placed it at 4°C at 10 times Dialyze in a normal saline solution of 10 volumes, change the dialysate every 3 hours, a total of three times; transfer to 10 volumes of 0.05mol / L boric acid solution for dialysis, change the dialysate every 3 hours, a total of four times; Put the liquid into a clean container and heat inactivate at 56°C for 1 hour; add 0.05% Proclin300 and 0.05% gentamycin, mix well, and then filter and sterilize with a 0.22 μm filter membrane to obtai...

Embodiment 2

[0051] A preparation method of a soluble ST2 enzyme-linked immunoassay kit, comprising the following steps: 1) Preparation of soluble ST2 standard and quality control products: take an appropriate amount of finely ground ammonium sulfate and add it while stirring at room temperature (18-25° C.) Fetal bovine serum to a final concentration of 200g / L, placed at 4°C for 20 hours; centrifuged at 10,000 rpm for 30 minutes, and filtered the supernatant with filter paper; put the filtered supernatant into a dialysis bag, and placed it at 4°C at 10 times dialysate in a normal saline solution of 10 volumes, and change the dialysate every 4 hours, a total of three times; transfer to 10 volumes of 0.05mol / L boric acid solution for dialysis, and change the dialysate every 4 hours, a total of four times; Put the liquid into a clean container and heat inactivate at 56°C for 1.5 hours; add 0.1% Proclin300 and 0.1% gentamycin, mix well, and then filter and sterilize with a 0.22 μm filter membra...

Embodiment 3

[0063] A preparation method of a soluble ST2 enzyme-linked immunoassay kit, comprising the following steps: 1) Preparation of soluble ST2 standard and quality control products: take an appropriate amount of finely ground ammonium sulfate and add it while stirring at room temperature (18-25° C.) Fetal bovine serum to a final concentration of 300g / L, placed at 4°C for 22 hours; centrifuged at 10,000 rpm for 40 minutes, and filtered the supernatant with filter paper; put the filtered supernatant into a dialysis bag, and placed it at 4°C at 10 times Dialyze in normal saline with 10 volumes, change the dialysate every 5 hours, three times in total; transfer to 10 volumes of 0.05mol / L boric acid solution for dialysis, change the dialysate every 5 hours, four times in total; Put the liquid into a clean container and heat inactivate at 56°C for 2 hours; add 0.15% Proclin300 and 0.15% gentamycin, mix well, and then filter and sterilize with a 0.22 μm filter membrane to obtain the fetal ...

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Abstract

The invention discloses a soluble ST2 enzyme-linked immunoassay kit. The immunoassay kit comprises a sample dilution liquid, a washing liquid, a coloring liquid, a stopping liquid and an enzyme label, the enzyme label is a horseradish peroxidase labeled soluble ST2 monoclonal antibody, the immunoassay kit also comprises multiple soluble ST2 standard substances with different concentrations, a soluble ST2 quality control substance and a microplate, and the microplate is coated with a soluble ST2 monoclonal antibody. The soluble ST2 standard substances with different concentrations can be directly used without dilution, so the soluble ST2 enzyme-linked immunoassay kit has the advantages of simplification of operating steps, shortening of the detection time, and realization of high-efficiency and rapid detection; the soluble ST2 quality control substance can be used to identify that a test result is accurate or not; and the soluble ST2 standard substances with different concentrations can be used to obtain assay kit with different specifications in order to adapt to use demands of different clients.

Description

technical field [0001] The invention belongs to the technical field of immunoanalysis medicine, and in particular relates to an enzyme-linked immunoassay kit for soluble ST2 and a preparation method thereof. Background technique [0002] Heart failure (HF) refers to a syndrome in which various cardiac structural or functional diseases lead to impaired ventricular filling and / or ejection ability, resulting in a series of clinical symptoms. According to the statistics of the World Health Organization, there are currently more than 22 million chronic heart failure patients worldwide, with more than 2 million new patients and more than 1 million deaths every year. The latest epidemiological survey results in China suggest that the prevalence of heart failure among urban and rural residents in my country is about 0.9%, and the prevalence of women is higher than that of men. Clinically, heart failure has an acute onset, serious condition, often atypical symptoms and signs, and ca...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/531G01N33/6893G01N2800/325
Inventor 兰成杰单存海陶剑
Owner TIANJIN KANGERKE BIOSCI