Biomarker, kit and detection method for liver cancer diagnosis

A technology of biomarkers and kits, applied in the field of molecular markers, can solve the problems of hepatocellular carcinoma plasma, etc., achieve high specificity, strong sensitivity, and improve the diagnostic rate

Active Publication Date: 2022-05-06
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there are no reports on exosomal protein markers and molecular classifiers derived from plasma of hepatocellular carcinoma.

Method used

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  • Biomarker, kit and detection method for liver cancer diagnosis
  • Biomarker, kit and detection method for liver cancer diagnosis
  • Biomarker, kit and detection method for liver cancer diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1, Extraction and Identification of Plasma Exosomes

[0092] Four plasma samples from hepatocellular carcinoma (HCC) patients and 4 healthy people (HC) were selected, matched according to age and sex, and plasma exosomes were isolated, and evaluated by nanoparticle size analysis, transmission electron microscope analysis, and Western blot analysis. Quality of extracted exosomes. The results of nanoparticle tracking analysis showed that the diameter of the enriched exosomes was about 30-150 nm, the peak particle size was about 72.25 nm, and the concentration was about 2.19 × 10 9 particles / mL ( figure 1 Middle A), similar in size to exosomes reported in the literature. Through transmission electron microscope analysis, it can be clearly observed that the size of plasma exosomes is between 40 and 150 nm ( figure 1 Middle B), the shape is like "saucer-shaped" or "oval", which is consistent with the morphology of exosomes. Western blot analysis showed that the ...

Embodiment 2

[0093] Example 2. Proteomic analysis of plasma exosomes

[0094] The proteomics map of liver cancer plasma exosomes was drawn, and the high-throughput mass spectrometry detection of the whole proteome was carried out by using the shotgun strategy and the data-independent acquisition (Data-independent acquisition, DIA) data acquisition method.

[0095] The present invention undergoes quality control filtering, data normalization and protein qualitative and quantitative analysis ( figure 2 Middle A), a total of 1434 proteins were identified. Among them, 20 proteins are known characteristic markers on the surface of exosomes, and their expression abundance ranks among the top among all identified proteins ( figure 2 Middle B). 1226 and 1346 proteins were identified in the HCC and HC groups, respectively ( figure 2 Middle C), there are 1138 shared proteins between the two groups.

[0096] The identified 1434 proteins were compared with exosome databases ExoCarta and Vesicle...

Embodiment 3

[0097] Example 3. Identification of candidate protein markers associated with liver cancer in plasma exosomes

[0098] The inventors performed principal component analysis (Principle component analysis, PCA). The results showed that the plasma exosomal proteome could significantly separate the HCC group from the HC group ( image 3 Middle A), suggesting that plasma exosomes undergo significant changes when liver cancer occurs.

[0099] In order to ensure the reliability of the analysis results, the inventors only retained the proteins detected in at least two samples, and used the KNN (K-nearest neighbor, k-Nearest Neighbor) method to fill in missing values. Finally, 1056 and 1186 proteins were retained in the HCC group and HC group, respectively, for subsequent analysis ( image 3 Middle B). Further, the inventors found that CD48 was the protein with the most significant difference between the HCC group and the HC group through protein differential expression analysis ( P...

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Abstract

The invention discloses a biomarker, a kit and a detection method for liver cancer diagnosis, belongs to the technical field of molecular markers, and aims to solve the technical problem of how to improve the sensitivity and / or specificity of hepatocellular carcinoma diagnosis. The invention provides an application of a biomarker and / or a substance for detecting the biomarker in hepatocellular carcinoma diagnosis or preparation of a product for hepatocellular carcinoma diagnosis. The biomarker is a plasma exosome CD48 protein or a combination of the plasma exosome CD48 protein and a plasma AFP (Alpha Fetoprotein). The biomarker CD48 protein and the detection method have the characteristics of strong sensitivity and high specificity, and the sensitivity can reach 0.928, so that the missed diagnosis rate (false negative rate) is low, the diagnosis rate of early liver cancer can be improved when the biomarker CD48 protein is applied to clinical diagnosis of hepatocellular carcinoma patients, sub-clinical liver cancer or latent liver cancer can be screened out from people, and the clinical application prospect is wide. The cure rate of liver cancer can be increased and the death rate can be reduced.

Description

technical field [0001] The invention belongs to the technical field of molecular markers for testing or molecular materials by means of chemical or physical properties of test materials, and specifically relates to a biomarker, a kit and a detection method for liver cancer diagnosis. Background technique [0002] Primary liver cancer (PLC) is a malignant tumor of the digestive system with the sixth highest incidence rate and the third highest mortality rate in the world. Hepatocellular carcinoma (HCC; hereinafter referred to as liver cancer) accounts for about 75%-85% of primary liver cancer. More than 60% of patients with hepatocellular carcinoma are diagnosed at an advanced stage, so timely and effective treatment cannot be obtained. At present, the sensitivity of serum markers commonly used in clinical diagnosis of liver cancer is low, so the identification of highly sensitive and specific diagnostic markers for liver cancer is one of the clinical problems that need to b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/68A61K45/00A61P35/00
CPCG01N33/57438G01N33/57476G01N33/57496G01N33/6893G01N33/6872A61K45/00A61P35/00G01N2333/70503G01N2333/471G01N2800/52
Inventor 周钢桥曹鹏博成意财杜振华冯岚
Owner ACADEMY OF MILITARY MEDICAL SCI
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