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Serum miRNA biomarker composition and application thereof

A biomarker and composition technology, applied in the field of biomarkers, can solve the problems of poor sensitivity of miRNA amplification, difficulty in detecting miRNAs, and analysis of PCR specificity, so as to achieve good sensitivity and specificity, improve reverse transcription efficiency, pain relieving effect

Active Publication Date: 2015-03-25
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Traditional miRNA detection techniques such as northern hybridization and microarray hybridization have poor amplification sensitivity to low-abundance miRNAs
Fluorescent quantitative PCR (qRT-PCR) is the most sensitive and reliable method for detecting gene expression, but conventional qRT-PCR technology is difficult to detect about 22bp miRNAs, in order to overcome this problem, Life Sciences and others have developed Taqman detection method, but The cost of this method is high, and the specificity of PCR response cannot be analyzed by solvent curve

Method used

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  • Serum miRNA biomarker composition and application thereof
  • Serum miRNA biomarker composition and application thereof
  • Serum miRNA biomarker composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, the collection of sample and the arrangement of sample data

[0038] A large number of serum samples were collected from the First Affiliated Hospital of Xiamen University. After sorting out the sample data, 9 cases of azoospermia were selected, aged 23-29 years, with an average age of 26±3 years. No sperm was found. There were 9 cases in the oligospermia group. The age of oligospermia patients was 22 to 30 years old, with an average age of 26±4 years old. 6 / mL. The control group consisted of 9 healthy male volunteers with a reproductive history, aged 24-29 years, with an average age of 26±3 years, and routinely normal semen.

Embodiment 2

[0039] Example 2. Preliminary screening of serum-specific miRNA expression chip for oligospermia and azoospermia

[0040] (1) Screening of chip samples

[0041] From the collected clinical serum samples, 9 cases of azoospermia were selected, aged 23 to 29 years, with an average age of 26±3 years, and no sperm was found in more than two routine semen examinations. There were 9 cases in the oligospermia group. The age of oligospermia patients was 22 to 30 years old, with an average age of 26±4 years old. 6 / mL. The control group consisted of 9 healthy male volunteers with a reproductive history, aged 24-29 years, with an average age of 26±3 years, and routinely normal semen.

[0042] (2) Collection of serum samples

[0043]Collect 2ml of peripheral blood from three groups of subjects to be tested, quickly transfer it into a blood collection tube, let it stand at room temperature for 15-30 minutes, centrifuge at 1900×g at 4°C for 10 minutes, carefully draw the supernatant into...

Embodiment 3

[0047] Example 3, Real-time fluorescence quantitative PCR method verification of oligospermia and azoospermia serum-specific miRNA

[0048] (1) Screening of verification samples

[0049] From the collected clinical serum samples, 20 cases of azoospermia were selected, aged 23 to 35 years, with an average age of 29±6 years. No sperm was found in more than two routine semen examinations. In the oligospermia group, there were 20 cases. The age of oligospermia patients was 22 to 34 years old, with an average age of 28±6 years old. 6 / mL. The control group consisted of 20 healthy male volunteers with a reproductive history, aged 24-36 years, with an average age of 30±6 years, and routinely normal semen.

[0050] (2) Collection of serum samples

[0051] Collect 2ml of peripheral blood from three groups of subjects to be tested, quickly transfer it into a blood collection tube, let it stand at room temperature for 15-30 minutes, centrifuge at 1900×g at 4°C for 10 minutes, carefull...

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Abstract

The invention relates to a biomarker, and particularly relates to a serum miRNA biomarker composition and an application thereof. The serum miRNA biomarker composition comprises at least one miRNA and sequences of reverse transcription primers, forward primers and reverse primers of all miRNAs. The serum miRNA biomarker composition can be used for preparing oligozoospermia and azoospermia detection reagents. An oligozoospermia and azoospermia detection kit comprises the serum miRNA biomarker composition, Taq enzyme, dNTP, MgCl2 and a PCR buffer solution. The serum miRNA biomarker composition provided by the invention has the advantages of high sensitivity, low sample consumption, wide detection range and wide linear quantitative range.

Description

technical field [0001] The present invention relates to biomarkers, in particular to a serum miRNA biomarker composition and application. Background technique [0002] miRNA, also known as microRNA, is a kind of single-stranded non-coding RNA composed of about 22 nucleotides, which mainly regulates the expression of target genes at the post-transcriptional level by direct cleavage of messenger RNA or indirect inhibition of translation. miRNAs have different expression patterns in different stages of individual development, such as cell differentiation, proliferation, apoptosis, etc., and in different tissues, indicating that they play an important regulatory role in development and differentiation. The study of miRNA expression profiles in tissues has found that miRNAs have strong cell, tissue or disease specificity, and these specifically expressed miNRAs are not only the basis for their functional studies, but also good disease markers. Studies in recent years have shown ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 曾骥孟李志明顾龙庄炫
Owner XIAMEN UNIV
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