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Application of hepatitis B virus pre-genome RNA in preparation of kit for predicting postoperative recurrence and postoperative survival of hepatitis B related liver cancer

A technology of survival prediction and kit, applied in the field of medical biological detection, can solve the problems of postoperative recurrence of hepatitis B-related liver cancer, unclear postoperative survival prediction, and no research report on HBVpgRNA, achieving high clinical reference value, high repeatability, good specific effect

Inactive Publication Date: 2021-08-10
THE NAVAL MEDICAL UNIV OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there is no research report on the function of HBV pgRNA in the occurrence and progression of hepatitis B-related liver cancer and its related molecular mechanism; in addition, whether serum HBV pgRNA can be used to evaluate postoperative recurrence of hepatitis B-related liver cancer and predict postoperative survival markers are not well defined and therefore deserve further exploration

Method used

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  • Application of hepatitis B virus pre-genome RNA in preparation of kit for predicting postoperative recurrence and postoperative survival of hepatitis B related liver cancer
  • Application of hepatitis B virus pre-genome RNA in preparation of kit for predicting postoperative recurrence and postoperative survival of hepatitis B related liver cancer
  • Application of hepatitis B virus pre-genome RNA in preparation of kit for predicting postoperative recurrence and postoperative survival of hepatitis B related liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Characterization Verification of HBV pgRNA

[0036] 1.1 Extraction of total RNA in serum

[0037] The present invention uses TRIzolTMLS Reagent (Thermo Fisher, product number: 10296010) to extract total RNA in patient serum. The experimental consumables used are all RNase-free. The general steps are: take 250ul serum, add 750ul TRIzolTMLS Reagent, shake and mix well, and let stand at room temperature for 5 minutes; add 200ul chloroform, shake vigorously for 15 seconds, let stand at room temperature for 10 minutes; centrifuge at 12000 rpm, 4 degrees Celsius for 15 minutes , move the supernatant to a new 1.5ml centrifuge tube, add an equal volume of isopropanol, and let stand at room temperature for 10 minutes; centrifuge at 12,000 rpm and 4°C for 10 minutes (white precipitate can be seen at the bottom of the tube); discard Liquid, save the precipitate, add 1ml of 75% ethanol, centrifuge at 10,000 rpm, 4°C for 5 minutes; discard the liquid, leave the precipit...

Embodiment 2

[0055] Example 2: Detection of the expression level of HBV pgRNA in serum of patients with hepatitis B-related liver cancer before operation

[0056] 2.1 Sample collection

[0057] A total of 136 preoperative sera of hepatitis B-related liver cancer patients who received NAs antiviral treatment for at least 1 year and whose serum HBV DNA was lower than the lower limit of clinical detection (less than 50 copies / mL) were collected and placed in a -80°C refrigerator. All cases had not received other treatment before the operation, all patients signed the informed consent, and the experimental protocol was approved by the ethics committee of the unit. The inclusion and exclusion criteria are shown in Table 1; the general clinical data are shown in Table 2.

[0058] Table 1. Inclusion and exclusion criteria for case selection

[0059]

[0060]

[0061] Table 2. General Clinical Data of Participants of the Invention

[0062]

[0063] Note: The degree of inflammation and ...

Embodiment 3

[0066] Example 3: The evaluation effect of serum HBV pgRNA on the prognosis of patients with hepatitis B-related liver cancer

[0067] Extraction, reverse transcription and real-time quantitative PCR of serum total RNA are as described in Example 1. We detected the expression of HBV pgRNA in serum of 136 patients before operation, and found that the overall survival rate of liver cancer patients with low serum HBV pgRNA expression was significantly higher than that of liver cancer patients with high serum HBV pgRNA expression ( image 3 A, Log-rank, P=0.0166), the cumulative recurrence rate of liver cancer patients with low serum HBV pgRNA expression was also significantly lower than that of liver cancer patients with high serum HBV pgRNA expression ( image 3 B, Log-rank, P=0.0157); The results of proportional hazard regression model univariate and multivariate analysis showed that the high expression of preoperative serum HBV pgRNA was associated with postoperative death (Ta...

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Abstract

The invention relates to the technical field of medical biological detection, in particular to an application of serum HBV pgRNA in preparation of a kit for predicting postoperative recurrence and postoperative survival of hepatitis B related liver cancer. The invention also provides a kit for predicting postoperative recurrence and postoperative survival of liver cancer by utilizing quantitative PCR detection of HBV pgRNA in preoperative serum of hepatitis B related liver cancer patients. The marker is high in clinical reference value and credibility, not only can predict liver cancer recurrence and postoperative survival conditions of hepatitis B related liver cancer patients after liver resection, but also can early recognize postoperative patients prone to recurrence, and intervention treatment can be carried out in advance. The kit and the detection method disclosed by the invention are simple, convenient, reliable, short in period, high in specificity and easy to clinically popularize.

Description

technical field [0001] The invention relates to the technical field of medical biological detection, in particular to the application of serum hepatitis B virus pregenomic RNA (HBV pregenomic RNA) in the preparation of postoperative recurrence and postoperative survival prediction kits for hepatitis B related liver cancer. Background technique [0002] Primary hepatocellular carcinoma (Hepatocellular Carcinoma, HCC, hereinafter referred to as liver cancer) is the most common malignant liver tumor in my country. Due to its high morbidity rate, rapid development, low surgical resection rate, easy occurrence of intrahepatic metastasis and recurrence, liver cancer has seriously affected the quality of life and health of the people. The proportion of liver cirrhosis and liver cancer caused by hepatitis B virus (Hepatitis B virus, HBV, hereinafter referred to as hepatitis B virus) infection is as high as 60% and 80% in my country. Therefore, chronic hepatitis B (Chronic Hepatitis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/70C12N15/11C12Q1/6851
CPCC12Q1/6851C12Q1/6886C12Q1/706C12Q2600/118C12Q2600/158C12Q2531/113C12Q2537/16C12Q2545/113
Inventor 丁文斌周伟平刘辉袁声贤杨富余建
Owner THE NAVAL MEDICAL UNIV OF PLA
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