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Preparation of hepatic fibrosis antibody protein chip and application of hepatic fibrosis antibody protein chip in fibrosis diagnosis staging

A liver fibrosis and protein chip technology, applied in disease diagnosis, analytical materials, biomaterial analysis, etc., can solve the problems of lack of serum liver fibrosis diagnostic indicators and limited role in liver fibrosis assessment, and achieves reduction of sampling difficulty and shortening Detection time, the effect of reducing the need

Pending Publication Date: 2020-06-26
HARBIN MEDICAL UNIVERSITY
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Problems solved by technology

[0005] At present, there is no large-scale serum sample bank of patients with liver fibrosis with different etiologies, and there is still a lack of serum-specific diagnostic indicators for liver fibrosis. At the same time, a single blood indicator has limited role in the assessment of liver fibrosis

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Preparation of liver fibrosis marker protein antibody chip detection system:

[0027] (1) Selection of the chip: select the glass slide as the film base, and silanize the slide glass.

[0028] (2) Preparation of antibody chip: Spot the monoclonal antibody of the above biomarker on a glass slide by dot matrix method, put the prepared chip into a wet box, and keep it overnight at 4°C.

[0029] Antibody protein chip detection serum samples:

[0030] (1) Labeling of serum samples: the serum samples were biotin-labeled for later use.

[0031] (2) Antibody chip detection of serum samples: The prepared antibody chip was sealed at room temperature for 1 hour, after washing, the serum sample was added to the chip, incubated with fluorescent dye, washed, and signal detection was performed with a fluorescent chip scanner. The obtained detection data is matched with the value measured by ELISA, and the corresponding fluorescence signal intensity of phase 0-4 is determined.

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Abstract

The invention discloses preparation of a hepatic fibrosis antibody protein chip and an application of the hepatic fibrosis antibody protein chip in fibrosis diagnosis staging. The method comprises thefollowing steps: establishing a sample library of serum biomarkers of liver fibrosis patients with different causes, screening biomarkers with higher sensitivity and specificity, preparing antibody protein chips for liver tissue biopsy staging, establishing a liver fibrosis marker protein antibody chip detection system, and applying the method to the fields of medical research and clinical medicine. According to the invention, the cause of liver fibrosis can be rapidly and efficiently detected by only taking peripheral blood of a patient, so the detection cost is reduced, the detection time is shortened, the requirement of liver lesion biopsy is reduced, the sampling difficulty is reduced, and the pain of the patient caused by sampling is relieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the preparation of a liver fibrosis antibody protein chip and its application in diagnosing fibrosis staging. Background technique [0002] Liver fibrosis is a common outcome of many chronic liver diseases, and biopsy is recognized as the "gold standard" for clinical diagnosis. However, due to its invasiveness and associated complications, the detection of serological markers is more attractive. A variety of biomarkers and predictive models for liver fibrosis have been found. However, various single biomarkers and combined scoring systems are biased towards fibrosis caused by different etiologies, and have their own advantages and disadvantages in sensitivity and specificity. Clinical work is more difficult. Therefore, establishing a sample bank of blood biomarkers in patients with liver fibrosis with different etiologies, screening biomarkers with high sensitivity and ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/552
CPCG01N33/6893G01N33/577G01N33/552G01N2800/085G01N2800/56G01N2800/60
Inventor 周洋王凯张书源张子谦
Owner HARBIN MEDICAL UNIVERSITY
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