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A kind of microribonucleic acid and application thereof for predicting interferon treatment chronic hbv curative effect

An interferon and chronic technology, applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., to save cost and time, easy to detect, and simplify the effect of primer libraries

Active Publication Date: 2017-02-15
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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  • A kind of microribonucleic acid and application thereof for predicting interferon treatment chronic hbv curative effect
  • A kind of microribonucleic acid and application thereof for predicting interferon treatment chronic hbv curative effect
  • A kind of microribonucleic acid and application thereof for predicting interferon treatment chronic hbv curative effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0061] The quantitative PCR method was used to find and prove that 10 hepatitis B patients whose interferon treatment is effective and 10 hepatitis B patients whose interferon treatment is not effective have stable presence of 5 differentially expressed human cytomegalovirus microribonucleic acids in serum / plasma. The specific steps are:

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

[0063] (2) Preparation of cDNA samples: a) Collect serum / plasma of hepatitis B patients whose interferon treatment is effective and ineffective; b) Take 100ml of serum from each patient, add 1ml of QIAzol lysis Reagent, and mix well; c) Phase separation: Place at room temperature for 5 minutes, then add 200ul chloroform to each portion, shake vigorously for 15 seconds, room temperature for 2-3 minutes, 12,000g, 4℃, cent...

Embodiment 2

[0069] Example 2 The application and preliminary evaluation of clinical value of the re-screened serum HCMV miRNA in the process of predicting the curative effect of interferon treatment of hepatitis B by quantitative PCR method after the sample was reintegrated and the sample volume was expanded.

[0070] The sera of 18 hepatitis B patients whose interferon treatment was effective and 18 hepatitis B patients whose interferon treatment failed were tested by qRT-PCR of microribonucleic acid. The experimental method and the qRT-PCR result processing method are the same as in Example 1.

[0071] The qRT-PCR results further selected two HCMV microRNAs that were significantly differentially expressed in the serum of hepatitis B patients who were effective with interferon treatment and those who did not. They were hcmv-mir-US4 and hcmv-mir-UL148D. ( Figure 3~4 ).

[0072] In order to preliminarily evaluate the effect of these two microRNAs in predicting the curative effect of interferon ...

Embodiment 3

[0075] Example 3 The double-blind experiment verifies the selected hcmv-miR-US4 as a biomarker to predict the efficacy of interferon in the treatment of hepatitis B.

[0076] QRT-PCR test of the hcmv-miR-US4 content in the serum of 102 patients with hepatitis B whose effects of interferon treatment are unknown. The experimental method and the qRT-PCR result processing method are the same as 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 treatment: 289.58 ( Figure 8 ). This value was used to analyze and classify the qRT-PCR results, and the predicted result was that 53 patients were ineffective with interferon treatment and 49 patients were effective in interferon treatment. Comparing the predicted results with the actual clinical results, the correct rate is calculated to reach 77.45%. Using the TaqMan probe of hcmv-mir-US4 sho...

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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 curative effect of interferon in treating chronic HBV and its application. Background technique [0002] Hepatitis B is an infectious disease with a worldwide distribution and serious harm. 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 are chronically infected, and about 1 million die each year from liver failure caused by HBV infection. Cirrhosis of the liver and primary liver cancer (HCC). There are currently about 93 million people with chronic HBV infection in my country, of which more than 20 million are patients with active hepatitis B who need treatment. In addition, HBV is highly infectious, and treatment of hepatitis B is undoubtedly very important. [0003] The current antiviral therapy for the treatment of chronic hepatitis B mainly re...

Claims

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Application Information

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Patent Type & Authority Patents(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