Kit for determining small-particle size homogeneous sol particle type cystatin C and preparation method therefor

A kit, particle type technology

Inactive Publication Date: 2011-08-10
WUHAN LIFE ORIGIN BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the short measurement time of the particle-enhanced immunoturbidimetric method, and it can be carried out on an automatic biochemical analyzer, it has become a routine application technology for the detection of cystatin C. However, the above method will produce precipitation after the reaction, which is not conducive to the cleaning of the biochemical analyzer.

Method used

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  • Kit for determining small-particle size homogeneous sol particle type cystatin C and preparation method therefor
  • Kit for determining small-particle size homogeneous sol particle type cystatin C and preparation method therefor
  • Kit for determining small-particle size homogeneous sol particle type cystatin C and preparation method therefor

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] A kit for measuring cystatin C in homogeneous sol particle type with small particle size and a preparation method thereof.

[0015] 1. The preparation of reagent R1 of the present invention

[0016] Prepare according to conventional methods.

[0017] 2. The preparation of reagent R2 of the present invention

[0018] I. Preparation of Colloidal Gold Solution

[0019] (1) prepare 1% chloroauric acid and 10% trisodium citrate with ultrapure water;

[0020] (2) Ultrapure water prepares 500ml of tetrachloroauric acid per ten thousandths (20ml 1% chloroauric acid adds 480ml ultrapure water);

[0021] (3) Heat and stir until slightly boiled, and quickly add trisodium citrate (i.e. 4ml 10% trisodium citrate) with a mass ratio of 1.2:1 to chloroauric acid;

[0022] (5) Heat and stir until the color turns into wine red, continue heating and stirring for 5-8 minutes and stop;

[0023] (6) Use an ultraviolet spectrophotometer to detect the absorbance value at 500nm-560nm, and ...

Embodiment 2

[0031] Embodiment 2 working curve is measured

[0032] The dosage of the present invention: the dosage of the reagent R1 of the present invention and the reagent R2 of the present invention are 160ul and 40ul respectively, and the dosage of the sample is 2ul.

[0033] The present invention adopts the endpoint method for measurement: 160ul R1 is added to 2ul sample, 40ul reagent R2 is added after 5 minutes at 37°C, and the reading point is started, and the point is read again after 5 minutes of reaction to obtain the absorbance difference.

[0034] Make the standard curve of the present invention: adopt the standard product of the present invention (choose 6 points to calibrate, calibrator cystatin C content is respectively 0.0, 0.5, 1.0, 2.0, 4.0, 8.0 mg / ml), use Mindray BS300 automatic biochemical instrument, the main detection wavelength is 546nm, and the secondary wavelength is 660nm;

[0035] According to the above-mentioned assay steps, the standard curve (such as figur...

Embodiment 3

[0036] Embodiment 3 accuracy, precision measurement

[0037] Utilize the quality control product provided by China Jiuqiang Company, choose two concentrations (lower and higher concentration), measure with the test kit of the present invention, the result is as follows.

[0038] Table 1

[0039]

[0040] Correlation experiment: using the reagent of the present invention and a control reagent (cystatin C kit of a well-known international company). The mindray BS300 automatic biochemical analyzer was used to measure 30 human sera, and the correlation analysis was carried out on the measured values. See the test results figure 2 , the X and Y axes in the figure are measured values ​​(cystatin C content mg / L).

[0041] Measurement of precision: Randomly select a normal serum sample and a serum sample of a patient with kidney disease for 20 consecutive measurements, and calculate according to CV(%)=SD / Mean×100.

[0042] Table 2

[0043] determination

[0044] ...

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Abstract

The invention provides a kit for immunoassay determining for small-particle size homogeneous sol particle type cystatin C and a preparation method therefor. The kit can be used for detecting the content of cystatin C in serum or blood plasma. The particle diameter of colloidal gold of the kit for the immunoassay determining for the homogeneous sol particle type cystatin C is 26 to 32nm. The kit has the characteristics of high sensitivity and high stability.

Description

technical field [0001] The invention relates to a homogeneous sol particle-type immunoassay kit and a preparation method thereof, in particular to a small-diameter homogeneous sol particle-type cystatin C immunoassay kit and a preparation method thereof. Background technique [0002] In 1983, Anastasi et al. isolated and purified high-purity cysteine ​​protease inhibitor (cysteine ​​proteinase inhibitor, CPI) from egg white for the first time, which was later named cystatin C (CysC). Cystatin C is a cysteine ​​protease inhibitor, also known as γ-trace protein and γ-postglobulin, which is a non-glycosylated basic protein with a molecular weight of about 13KD. All cells can be produced, filtered by the glomerulus and almost all reabsorbed and decomposed by the proximal convoluted tubule, and the production rate is mostly in a relatively stable state. Cystatin C is a relatively ideal endogenous marker that reflects the rate of glomerular filtration. Studies have shown that thi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/53
Inventor 华权高沈鹤霄周许可
Owner WUHAN LIFE ORIGIN BIOTECH LTD
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