MicroRNA used for predicting curative effect of interferon on chronic HBV and application thereof

An interferon, chronic technology, applied in the direction of microbial determination/inspection, DNA/RNA fragmentation, recombinant DNA technology, etc., to achieve the effect of saving cost and time, easy detection, and simplifying the primer library

Active Publication Date: 2015-03-18
JIANGSU MICROMEDMARK BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recent studies have shown that serum / plasma microRNA can be used for the early diagnosis and treatment guidance of various cancers, but there is no report on the use of serum / plasma microRNA to predict the efficacy of interferon therapy in patients with chronic hepatitis B

Method used

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  • MicroRNA used for predicting curative effect of interferon on chronic HBV and application thereof
  • MicroRNA used for predicting curative effect of interferon on chronic HBV and application thereof
  • MicroRNA used for predicting curative effect of interferon on chronic HBV and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Preliminary screening of specific HCMV miRNA expression profiles in patients with chronic hepatitis B

[0061] Quantitative PCR method was used to discover and prove that there were 5 differentially expressed microRNAs of human cytomegalovirus stably in the serum / plasma of 10 hepatitis B patients who were effective in interferon therapy and 10 hepatitis B patients who were ineffective in interferon therapy. The specific steps are:

[0062] (1) Collect serum / plasma from hepatitis B patients who are effective and ineffective in interferon therapy

[0063] (2) Prepare cDNA samples: a) Collect the serum / plasma of hepatitis B patients who are effective and ineffective in interferon treatment; b) Take 100ml of serum from each patient, add 1ml of QIAzol lysis Reagent, and mix well; c) Phase separation: Leave at room temperature for 5 minutes, then add 200ul chloroform to each portion, shake vigorously for 15s, room temperature for 2-3 minutes, centrifuge at 12,000g,...

Embodiment 2

[0069] Example 2 After the samples were reintegrated and the sample size was enlarged, the quantitative PCR method was used to detect the re-screened serum HCMV miRNA and its application and clinical value preliminary evaluation in the process of predicting the curative effect of interferon treatment of hepatitis B.

[0070] The qRT-PCR test of microRNA was performed on the serum of 18 hepatitis B patients who were effective in interferon therapy and 18 hepatitis B patients who were ineffective in interferon therapy. The experimental method and qRT-PCR result processing method are the same as those in Example 1.

[0071] The results of qRT-PCR further selected two HCMV microRNAs that were significantly differentially expressed in the serum of hepatitis B patients with effective interferon treatment and those with ineffective interferon treatment, namely hcmv-mir-US4 and hcmv-mir-UL148D. ( Figure 3-4 ).

[0072]In order to preliminarily evaluate the effect of these two micro...

Embodiment 3

[0075] Example 3 The double-blind experiment verifies the effect of the screened hcmv-miR-US4 as a biomarker in predicting the curative effect of interferon in the treatment of hepatitis B.

[0076] qRT-PCR test of hcmv-miR-US4 content in the serum of 102 cases of hepatitis B patients with unknown interferon treatment effect. The experimental method and qRT-PCR result processing method are the same as those in Example 1.

[0077] According to the results of Example 2, we used quantitative PCR to calculate the value of hcmv-miR-US4 content in the serum of patients with effective and ineffective interferon therapy: 289.58 ( Figure 8 ). And use this value to analyze and classify the qRT-PCR results, the prediction result is that 53 patients are ineffective with interferon treatment, and 49 patients are effective with interferon treatment. Comparing the predicted results with the actual clinical results, the correct rate reaches 77.45%. Using the TaqMan probe of hcmv-mir-US4 s...

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Abstract

The invention belongs to the technical field of biology, and relates to a microRNA (miRNA) used for predicting curative effect of interferon on chronic HBV and application thereof. The serum miRNA combination is mainly composed of two kinds of mature miRNA such as human cytomegalovirus-derived hcmv-mir-US4 and hcmv-mir-UL148D. The screened-out HCMV miRNA combination has the specificity and the sensitivity of predicting the curative effect of using interferon to treat chronic viral hepatitis B patients, and the specificity and the sensitivity are substantially higher than those of current clinic experience prediction, and thus the prediction accuracy is improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a microribonucleic acid for predicting the therapeutic effect of interferon on chronic HBV and its application. Background technique [0002] Hepatitis B is a serious infectious disease with worldwide distribution. According to the World Health Organization, about 2 billion people in the world have been infected with hepatitis B virus (HBV), of which 350 million people are chronically infected, and about 1 million people die every year from liver failure caused by HBV infection. Cirrhosis and primary liver cancer (HCC). There are about 93 million chronic HBV-infected people in my country, among which there are about 20 million active hepatitis B patients who have symptoms and need treatment. Coupled with the strong infectivity of HBV, the treatment of hepatitis B is undoubtedly very important. [0003] The current antiviral therapy for the treatment of chronic hepatitis B mainly rel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/158C12Q2600/178
Inventor 曾科潘怡王楠李丽民张辰宇
Owner JIANGSU MICROMEDMARK BIOTECH
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