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
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
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...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 


