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Autoantibody 7-AAb detection panel for hepatocellular carcinoma and application of autoantibody 7-AAb detection panel

A technology of hepatocellular carcinoma and autoantibodies, which is applied in the field of autoantibody 7-AAb detection panel of hepatocellular carcinoma, can solve the problems of non-specificity and achieve good diagnostic efficiency

Pending Publication Date: 2020-07-14
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have found that there are some tumor-specific antigens, but no specificity among different tumors, such as p53; there are also some tumor-specific antigens, such as many antigens in prostate cancer are oxidative stress-related proteins

Method used

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  • Autoantibody 7-AAb detection panel for hepatocellular carcinoma and application of autoantibody 7-AAb detection panel
  • Autoantibody 7-AAb detection panel for hepatocellular carcinoma and application of autoantibody 7-AAb detection panel
  • Autoantibody 7-AAb detection panel for hepatocellular carcinoma and application of autoantibody 7-AAb detection panel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 HuProt TM Human proteome microarray detects liver cancer and healthy control sera

[0033] see process figure 1 Shown, specifically include the following steps

[0034] (1) Use 1 HuProt for each serum sample TM The proteome chip is used for detection, specific antibodies (including IgG, IgM) bind to the protein immobilized on the chip, wash to remove unbound antibodies and other proteins, and then use anti-human IgM fluorescently labeled secondary antibody (cy5 labeling, showing Red) and anti-human IgG fluorescent secondary antibody (cy3-labeled, green) detection, the signal is read by a fluorescent scanner, and the strength of the signal is positively correlated with the affinity and quantity of the antibody.

[0035] (2) A total of 50 cases of liver cancer and 50 cases of healthy control serum were detected. In order to eliminate errors caused by systematic differences between different samples and different chips, intra-chip and inter-chip normalization...

Embodiment 2

[0036] Example 2 Construction of HCC Focused Arrays

[0037](1) On the basis of normalization, statistical analysis was performed on the data to screen out specific high-response proteins of the liver cancer group. The analysis logic is as follows:

[0038] Parametric test t test, p-value<0.05 means that there is a significant difference between the two;

[0039] Calculate the multiple of difference between groups, when fold change ≥ 1.2, it is considered that there is a potential difference between the two;

[0040] Set cut off=mean+2SD (99%CI) for healthy group samples, and calculate the positive rate of HCC, with 10% as the minimum screening standard;

[0041] A total of 100 candidate biomarkers with certain discrimination ability or combined discrimination ability were screened out.

[0042] For the construction of HCC Focused Arrays and the list of 100 human recombinant proteins, see figure 2 .

[0043] (2) Use these 100 human recombinant proteins to make HCC Focused...

Embodiment 3

[0044] Example 3 Autoantibody 7-AAb Detection Panel Screening Process

[0045] Based on the small protein chip prepared in Example 2, we detected 561 cases of liver cancer patients and 592 cases of controls (including 343 cases of normal NC and 249 cases of LC of liver cirrhosis patients). Among them, 576 samples (282 liver cancer patients, 164 normal patients, and 130 liver cirrhosis patients) participated in model training (training set), and 577 samples (279 liver cancer patients, 179 normal patients, and 119 liver cirrhosis patients) were used as independent samples The model is validated by blind testing (validation set).

[0046] 1. Sample testing

[0047] 1) Rewarming: Take the chips out of the -80°C refrigerator, rewarm in a 4°C refrigerator for half an hour, and then rewarm at room temperature for 15 minutes.

[0048] 2) Sealing: After rewarming the chip, fix 14blocks fences, after fixing, add blocking solution to each block, place on a side swing shaker, and seal a...

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Abstract

The invention belongs to the technical field of liver cancer screening, and particularly relates to an autoantibody 7-AAb detection panel for hepatocellular carcinoma and application of the autoantibody 7-AAb detection panel. According to the invention, 561 hepatocellular carcinoma patients and 592 control (including 343 normal human NC and 249 liver cirrhosis patients LC) samples are detected; according to the present invention, the autoantibodies of the seven proteins such as CIAPIN1, EGFR, MAS1, SLC44A3, ASAH1, UBL7 and ZNF428 are determined to have the liver cell liver cancer distinguishing ability through the 10-cold cross-validation; by applying the seven molecules and establishing a neural network (ANN) model, and it is found that the ANN model shows better diagnostic efficiency andis superior to a traditional logistic regression model and an alpha fetoprotein (AFP). In a verification set, the ANN model based on the seven molecules also shows good diagnostic efficiency, especially sensitivity.

Description

technical field [0001] The invention belongs to the field of liver cancer detection, and in particular relates to an autoantibody 7-AAb detection panel for hepatocellular carcinoma and an application thereof. Background technique [0002] Hepatocellular carcinoma (Hepatocellular carcinoma, referred to as: HCC) is one of the common malignant tumors, its onset is occult, the degree of malignancy is high, and the mortality rate is high. Therefore, timely and accurate diagnosis is very important to improve the survival rate of patients. At present, alpha-fetoprotein (AFP) combined with imaging and pathological examinations is mainly used for early diagnosis of liver cancer clinically; however, the specificity and sensitivity of AFP for liver cancer screening are not very satisfactory. With the continuous development of molecular biology, a variety of new markers have been discovered, especially the diagnostic value of autoantibodies. [0003] Autoantibodies are specific antibod...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/564G16B40/00
CPCG01N33/57438G01N33/57484G01N33/564G16B40/00
Inventor 樊嘉高强张舒毕利军李阳刘羽鸣杨思贤刘诚喜
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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