Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for aldosterone, and preparation method of kit

A chemiluminescence immunoassay and quantitative detection technology, applied in the field of immunoassay medicine, can solve the problems of short validity period of reagents, narrow detection range, complicated operation, etc., and achieve the effects of good stability, simple operation and high sensitivity.

Inactive Publication Date: 2013-03-27
BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The problem to be solved by the present invention is to provide ALD magnetic particle chemiluminescence immunoquantitative detection kit and its preparation method, which avoids the shortcomings of radioimmunoassay reagents such as short validity period, radioactive pollution, cumbersome operation, etc., and solves the problem of low sensitivity and detection Narrow scope, high cost defects

Method used

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  • Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for aldosterone, and preparation method of kit
  • Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for aldosterone, and preparation method of kit
  • Quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for aldosterone, and preparation method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Preparation of Aldosterone (ALD) Magnetic Particle Chemiluminescent Immunoquantitative Assay Kit I

[0033] (1) Preparation of aldosterone calibrator:

[0034] Dilute the pure ALD product with phosphate buffer containing 50% bovine serum to form a series of gradients, the concentrations are 0, 60, 150, 450, 1500, 4000 pg / mL.

[0035] (2) Preparation of aldosterone quality control products:

[0036] The ALD pure product was prepared with a 50% bovine serum calibrator dilution to prepare a low-value quality control product and a high-value quality control product, and the concentrations were 196pg / mL and 2820pg / mL, respectively.

[0037] (3) Preparation of magnetic particle-streptavidin suspension:

[0038] Take 100mL0.1mol / L 2-morpholineethanesulfonic acid solution (MES solution), add 10mg magnetic particles and 3mg streptavidin (SA) in turn, stir for 30min, then add 10mg / mL carbodiimide hydrochloride solution (1-Ethyl-(3-dimethylaminopropyl, EDC) 3.5uL, afte...

Embodiment 2

[0049] Example 2: Preparation of Aldosterone (ALD) Magnetic Particle Chemiluminescent Immunoquantitative Assay Kit II

[0050] (1) With the preparation of embodiment 1 aldosterone calibrator

[0051] (2) Preparation of aldosterone quality control products:

[0052] The ALD pure product was prepared with a 50% bovine serum calibrator dilution to prepare a low-value quality control product and a high-value quality control product, and the concentrations were 294pg / mL and 1880pg / mL, respectively.

[0053] (3) Preparation of the magnetic particle-streptavidin suspension as in Example 1

[0054] (4) Preparation of biotin-labeled aldosterone antibody

[0055] Take 0.5 mg of aldosterone antibody and dialyze with borate buffer solution at 2-8°C for 2 hours; add 25 μg of biotin to the dialyzed antibody and add dimethyl sulfoxide at the same time, the final concentration is 10%, and react in the dark for 3 hours. Shake slowly; add 250uL 1mol / L ammonium chloride solution to the above ...

Embodiment 3

[0057] Example 3: Preparation of Aldosterone (ALD) Magnetic Particle Chemiluminescent Immunoquantitative Assay Kit III

[0058] (1) With the preparation of embodiment 1 aldosterone calibrator

[0059] (2) Preparation of aldosterone quality control products:

[0060] The ALD pure product was prepared with a 50% bovine serum calibrator dilution to prepare a low-value quality control product and a high-value quality control product, and the concentrations were 205pg / mL and 2540pg / mL, respectively.

[0061] (3) Preparation of the magnetic particle-streptavidin suspension as in Example 1

[0062] (4) Preparation of biotin-labeled aldosterone antibody

[0063] Take 0.5 mg of aldosterone antibody and dialyze it with borate buffer at 2-8°C for 3 hours; add 25 μg of biotin to the dialyzed antibody and add dimethyl sulfoxide at the same time, the final concentration is 10%, and react in the dark for 3 hours. Shake slowly; add 250uL 1mol / L ammonium chloride solution to the above solut...

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Abstract

The invention discloses a quantitative detection kit combining magnetic particles with chemiluminescence immunoassay for aldosterone. The kit comprises aldosterone calibrators, magnetic particle suspension coupled with streptavidin, an aldosterone antibody labeled with biotin, an aldosterone enzyme combination, an aldosterone quality controller, chemiluminescence liquor A, chemiluminescence liquor B, 20 times concentrated washing liquor, and a reaction tube, wherein enzyme adopted by the aldosterone enzyme combination is horse radish peroxidase with the purity RZ being more than or equal to 3.0 and the activity being more than or equal to 250U / ml. The invention also discloses a preparation method of the kit. Compared with the conventional kit, the quantitative detection kit is simple and convenient to operate, is safe, does not cause environment pollution, and also has the advantages of wide concentration range, low cost, good stability and the like of detection samples.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention provides an aldosterone (ALD) magnetic particle chemiluminescent immunoquantitative detection kit and a preparation method thereof. Background technique [0002] Hypertension is the most common cardiovascular disease and a major public health problem worldwide. According to the latest data from the Ministry of Health, there are more than 200 million hypertensive patients in my country, and hypertension ranks first among the top ten chronic diseases, and it is increasing year by year. Hypertension is a major risk factor for hypertensive heart disease, coronary heart disease, heart failure, chronic renal failure, and cerebrovascular disease. Clinically, aldosterone, angiotensin Ⅰ, angiotensin Ⅱ, and norepinephrine are of great significance to the diagnosis and classification of hypertension, as well as the assessment of the severity and prognosis of coronary heart...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/531G01N21/76
Inventor 刘萍张影宋启超范利花
Owner BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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