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Diagnostic kit for liver fibrosis or liver cirrhosis

A technology of liver fibrosis and kits, which can be applied in the fields of disease diagnosis, biological testing, material inspection, etc., can solve the problems of large damage to patients, missed diagnosis, long detection cycle, etc., and achieve the effect of high sensitivity, simple usage, and small damage

Active Publication Date: 2020-04-03
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pathological detection is the gold standard for the diagnosis of liver fibrosis, but it is prone to false negatives, because the distribution of liver fibrosis caused by many diseases is uneven, and if no samples of the fibrosis site are obtained by liver puncture, the diagnosis will be missed; in addition, the liver The puncture is more harmful to the patient and the detection cycle is longer

Method used

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  • Diagnostic kit for liver fibrosis or liver cirrhosis
  • Diagnostic kit for liver fibrosis or liver cirrhosis
  • Diagnostic kit for liver fibrosis or liver cirrhosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 Kit of the present invention (flow cytometry detection kit)

[0031] 1. Kit composition

[0032] Core components: Fluorescence-labeled antibodies to Ceacam1 and CSF1R (primary fluorescent antibodies).

[0033] Other components (can be prepared by yourself): monocyte isolation and extraction kit (commercially available).

[0034] 2. How to use

[0035] 1) Take the peripheral blood of the subject to be tested, and separate the peripheral blood mononuclear cells (monocyte separation and extraction kit can be used);

[0036] 2) Suspend monocytes in PBS, add fluorescent primary antibody to incubate;

[0037] 3) Use flow cytometry to detect antibody fluorescence. If the corresponding marker fluorescence of Ceacam1 and CSF1R is detected, and the intensity is significantly higher than that of healthy people, it indicates that the subject has liver fibrosis.

[0038] This kit does not detect Mac-1, because the sample taken is peripheral blood, and Mac-1 is a wel...

Embodiment 2

[0039] Embodiment 2 kit of the present invention (liver tissue paraffin section immunofluorescence detection kit)

[0040] 1. Composition of the kit

[0041] Core components: Mac-1, Ceacam1, CSF1R fluorescently labeled antibodies (fluorescent primary antibodies).

[0042] Other components (can be prepared by yourself): Antigen retrieval solution (Epitope retrieval solution), blocking solution (goat serum), washing buffer (commercially available for immunohistochemistry).

[0043] 2. How to use

[0044] 1) Paraffin sections of liver tissue were taken and placed in xylene-xylene-xylene-100% ethanol-95% ethanol-85% ethanol-75% ethanol-70% ethanol-double distilled water for dewaxing hydration (each solvent 3min);

[0045] 2) Antigen retrieval: Put the dewaxed and hydrated liver tissue slices into a container, add antigen retrieval solution, and microwave for 8 minutes, twice;

[0046] 3) Sealing: naturally lower to room temperature, wash with double distilled water 3 times, an...

experiment example 1

[0051] Experimental example 1 Detection of PFDM in the liver of fibrotic mice

[0052] 1. Method

[0053] Use immunofluorescence to detect the distribution of PFDM in liver tissue of mice with liver fibrosis and healthy control mice, the steps are as follows:

[0054] 1) The liver tissues of control / hepatic fibrosis mice were taken and fixed in formalin for 24 hours;

[0055] 2) after dehydration and transparency, embedding in paraffin and sectioning to obtain paraffin sections;

[0056] 3) Mac-1, Ceacam1, and CSF1R were detected according to Example 2.

[0057]2. Results

[0058] The results showed that there was no PFDM in the liver of normal mice, but a large amount of PFDM was distributed in the fibrous septa of the liver of fibrotic mice ( figure 1 ).

[0059] Conclusion: PFDM in the liver is related to liver fibrosis; people can detect liver fibrosis by detecting PFDM in the liver.

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Abstract

The invention belongs to the field of disease diagnosis kits, and particularly discloses a liver fibrosis and liver cirrhosis diagnosis kit which comprises a reagent for detecting Ceacam1 + CSF1R + peripheral blood mononuclear cells. Experiments show that the content of Ceacam1 + CSF1R + peripheral blood mononuclear cells relative to healthy people can reflect whether a tested object has liver fibrosis or liver cirrhosis or not, and the sensitivity of the Ceacam1 + CSF1R + peripheral blood mononuclear cells is higher than that of a current liver function detection index (ALT / AST). The kit provided by the invention has the advantages of few detection indexes, high detection sensitivity and good application prospect.

Description

technical field [0001] The invention belongs to the field of disease diagnosis reagents. Background technique [0002] Liver fibrosis is a pathological process in which extracellular matrix accumulation during tissue regeneration after liver structure is damaged leads to fibrous nodules; the final stage of its development is liver cirrhosis, which is one of the common chronic progressive liver diseases and major fatal diseases in clinic one. Every year in my country, 2% to 3% of liver fibrosis patients will develop into liver cirrhosis, accounting for 4.3% to 14.2% of the total number of inpatients in internal medicine, and the number of people who die of liver cirrhosis and its complications is as high as 300,000, which brings great harm to the country and the people. heavy economic and social burdens. Therefore, the early diagnosis and screening of liver fibrosis is of great significance to curb the further development of liver fibrosis into liver cirrhosis. [0003] The...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893G01N33/56972G01N2800/085Y02A50/30
Inventor 夏杰黄爱龙刘迪娜唐霓汪凯蔡莹沙雨潘敏刚
Owner CHONGQING MEDICAL UNIVERSITY
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