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A kind of cystatin c detection kit and preparation method thereof

A technology for detecting kits and cystatin, which is applied in the field of medical immunology in vitro diagnosis, and achieves the effects of high sensitivity, improved sensitivity and high accuracy

Active Publication Date: 2015-08-05
浙江夸克生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, latex particle-enhanced turbidimetry is the most common method, but the minimum detection limit of latex particle-enhanced turbidimetry can only reach 0.1 mg / l, and the linearity can only reach 8 mg / l, which has limitations in clinical application.

Method used

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  • A kind of cystatin c detection kit and preparation method thereof
  • A kind of cystatin c detection kit and preparation method thereof
  • A kind of cystatin c detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Preparation of reagent R1: add corresponding weight of PEG6000 and sodium chloride to Tris buffer and mix well, then add NaN 3 , adjust the pH value; the components of reagent R1 are:

[0029]

[0030] Preparation of reagent R2: add CDI to activate polystyrene latex for 12 minutes, centrifuge at 18000g / min for 25min, discard supernatant, add PB buffer to dissolve, add goat anti-human cystatin C polyclonal antibody and shake at room temperature for 1 hour, 18000g / min Centrifuge for 25 minutes, discard the supernatant, and add 0.01mol / L Tris buffer to get it; the components of reagent R2 are:

[0031]

[0032] Preparation of calibrator: Remove corresponding amounts of cystatin C and add them to 0.01mol / L Tris buffer to obtain six gradient calibrator with final concentrations of 0.1, 0.5, 1.0, 2.0, 4.0 and 8.0 mg / L .

[0033] Pack the above reagents separately to obtain the cystatin C detection kit

Embodiment 2-5

[0035] Except each reagent component, other is all with embodiment 1, and concrete component is as follows:

[0036] Reagent R1 consists of:

[0037]

[0038] Reagent R2 consists of:

[0039]

Embodiment 6

[0041] The minimum detection limit test of the detection kit prepared by embodiment 1-5

[0042] Adopt Hitachi 7080 automatic biochemical analyzer and the kit of the present invention to measure the human serum sample and blank solution (physiological saline or purified water) of known concentration, the specific method is:

[0043]

[0044] After mixing and incubating at 37°C for 30 seconds, read the absorbance A1, and after continuing to incubate for 5 minutes, read the absorbance A2, and calculate ΔA=A2-A1.

[0045] The analysis method is: two-point endpoint method, the main wavelength is 546nm, and the secondary wavelength is 700nm.

[0046] The sensitivity detection result of the kit prepared in Example 1 is as follows:

[0047]

[0048] The minimum detection limit of the kit prepared by the present invention is: [Microsoft user 1]

[0049] The minimum detection limit of the kit prepared by embodiment 2-5 is shown in the following table:

[0050]

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Abstract

The invention relates to the field of medical immunology in vitro diagnosis, and especially provides a detection kit for detecting cystatin C in serum. The detection kit comprises a reagent R1, a reagent R2 and calibration materials. The ingredients of the reagent R1 are a Tris buffer with a concentration of 0.01-0.05mol / L, PEG 6000 with a concentration of 10-40g / L, NaN3 with a concentration of 0.5-1.5g / L, and sodium chloride with a concentration of12-20g / L, and the pH value is 8.0-8.5. The ingredients of the reagent R2 are a Tris buffer with a concentration of 0.01-0.02mol / L, and polystyrene latex granules coated by goat-anti-human cystatin C polyclonal antibodies with a concentration of 0.5-3.0 g / L. The calibration materials are six gradient standards with cystatin C and the contents of cystatin C are 0.1, 0.5, 1.0, 2.0, 4.0 and 8.0mg / L. The solution is a Tris buffer with a concentration of 0.01mol / L. The kit has simple composition, good stability and high accuracy.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis of medical immunity, in particular to a detection kit for detecting cystatin C in serum. Background technique [0002] Kidney disease is a common clinical disease, and renal dysfunction caused by various reasons is a risk factor for the progression to end-stage renal disease and cardiovascular disease. The only way to stop kidney disease from getting worse is to diagnose it early and treat it to reverse damaged kidney function. [0003] Cystatin C is a non-glycosylated protein, a class of cysteine ​​protease inhibitors, which is produced in a constant manner by most nucleated cells. Its molecular weight is small, and its production rate is stable, because its concentration in serum is closely related to glomerular filtration rate (GFR), and it is more sensitive to glomerular filtration rate than creatinine. Therefore, cystatin C is of great significance to the determination of glomerular filtra...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/54393G01N33/544G01N33/6803G01N2333/8139
Inventor 陈青松张伟林韩钟
Owner 浙江夸克生物科技有限公司
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