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Detection reagent for human CK18 protein and preparation method of reagent

A technology for detecting reagents and reagents, applied in the field of laboratory medicine, can solve the problems of poor control of sample dilution, long reaction measurement time, and narrow linear range, and achieve the effects of improved accuracy, simple operation and wide linear range.

Inactive Publication Date: 2018-07-10
苏州和锐生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ELSA has high sensitivity, but its operation is complicated, the reaction measurement time is long, and the linear range is narrow, so it is difficult to control the sample dilution; chemiluminescence has the advantage of high sensitivity, but it needs to be used with special equipment, which is expensive and not conducive to Promoted by hospitals at all levels, and the standard curve will drift over time

Method used

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  • Detection reagent for human CK18 protein and preparation method of reagent
  • Detection reagent for human CK18 protein and preparation method of reagent
  • Detection reagent for human CK18 protein and preparation method of reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 CK18 Latex Enhanced Immunoturbidimetric Detection Reagent 1

[0033] 1. The main components and concentrations of the kit are as follows:

[0034] Reagent R1:

[0035]

[0036] Reagent R2:

[0037]

[0038] Latex particles coated with mouse anti-human CK18 monoclonal antibody and chicken anti-human CK18 polyclonal antibody Latex particle size: 160nm, latex concentration 0.2%.

[0039] CK18 calibrator:

[0040]

[0041] Latex particles coated with mouse anti-human CK18 monoclonal antibody and chicken anti-human CK18 polyclonal antibody Latex particle size: 160nm, latex concentration 0.2%.

[0042] CK18 calibrator:

[0043]

[0044]

[0045]Add CH18 pure product to the above buffer solution according to the required concentration of CK18 reference calibrator, prepare a reference calibrator with a concentration of 3200U / L, and then serially dilute with calibrator diluent to obtain 6 concentrations of 3200U / L and 1600U / L , 800U / L, 400U / L, 200U / ...

Embodiment 2

[0057] Example 2 CK18 Latex Enhanced Immunoturbidimetric Detection Reagent 2

[0058] 1. The main components and concentrations of the kit are as follows:

[0059] Reagent R1:

[0060]

[0061] Reagent R2:

[0062]

[0063] Sensitized latex particles coated with mouse anti-human CK18 monoclonal antibody and rabbit anti-human CK18 polyclonal antibody, latex particle size: 160nm; latex concentration 0.2%.

[0064] CK18 calibrator:

[0065]

[0066] Add CH18 pure product to the above buffer solution according to the required concentration of CK18 reference calibrator, prepare a reference calibrator with a concentration of 3200U / L, and then serially dilute with calibrator diluent to obtain 6 concentrations of 3200U / L and 1600U / L , 800U / L, 400U / L, 200U / L, 0U / L series of reference calibration products.

[0067] Reagent R1 is a colorless transparent liquid, R2 is a milky white liquid, and the calibrator is a light yellow transparent solution.

[0068] 2. Kit assay metho...

Embodiment 3

[0078] Example 3 Performance Evaluation of CK18 Latex Enhanced Immunoturbidimetric Detection Reagent

[0079] 1. Correlation Linear Evaluation

[0080] Correlation experiments were carried out between the kit 1 of the present invention prepared according to Example 1 and the commercially available kit A measured by enzyme-linked immunosorbent assay (ELISA). Detect 50 fresh human sera (including normal and abnormal samples), measure according to their respective determination methods, and perform correlation analysis on the measured values ​​(results see figure 1 , X and Y axes are measured values, unit U / L). Correlation coefficient R 2 =0.9927, the regression equation is y=1.0145x+19.915, the result shows that the reagent of the present invention has a good correlation with ELISA. The correlation experiment data are shown in Table 1.

[0081] The kit 2 of the present invention prepared according to Example 2 and the commercially available imported kit B were tested for cor...

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Abstract

The invention relates to a detection reagent for CK18 and a preparation method, in particular to a reagent for detecting the CK18 by adopting latex-enhanced immunoturbidimetry and a preparation method. The reagent includes a reagent R1, a reagent R2 and a calibration product, wherein the reagent R1 contains a reaction accelerant, a preservative, a surfactant, a stabilizer, an electrolyte and a buffer solution; the reagent R2 contains latex particles combined with anti-CK18 monoclonal antibodies and polyclonal antibodies, a preservative, a surfactant, a stabilizer, an electrolyte and a buffer solution; the calibration product contains a preservative, an electrolyte, a stabilizer, a CK18 pure product and a buffer solution. According to the detection reagent for the CK18 and the preparation method, by adopting the method of using the monoclonal antibodies and the polyclonal antibodies to be combined with and coat the latex particles, the high sensitivity and wide linearity range of a reagent kit are ensured, and meanwhile the detection reagent for the CK18 and the preparation method also have the advantages of high accuracy, good repeatability, high specificity, simple operation and the like, and can be applied to a clinical commonly-used full-automatic biochemical analyzer.

Description

technical field [0001] The invention relates to a cytokeratin 18 (CK18) detection reagent and a preparation method, in particular to a reagent and a preparation method for detecting CK18 by using latex-enhanced immune turbidimetry, which can be applied in the field of laboratory medicine. Background technique [0002] CK18 is a kind of cytoskeleton protein. In addition to supporting the cytoskeleton, it also participates in various reactions in the cell through various channels. As an indicator of hepatic cell apoptosis, CK18 is elevated in many liver diseases including non-alcoholic fatty liver disease (NAFLD), and is associated with the degree of steatosis, lobular inflammation, ballooning, and fibrous changes in liver cells in NAFLD patients. The level of serum CK18 in patients with moderate to severe fibrosis was significantly higher than that in patients with mild or no fibrosis. [0003] Liver disease is a major problem that threatens human health. The incidence of fa...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/68
Inventor 陈菲周鹏飞霍如松
Owner 苏州和锐生物科技有限公司
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