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A kind of serum miRNA biomarker composition and application

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: 2017-08-29
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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  • A kind of serum miRNA biomarker composition and application
  • A kind of serum miRNA biomarker composition and application
  • A kind of serum miRNA biomarker composition and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Collection of samples and 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 from 23 to 29 years old, with an average age of 26±3 years old. No sperm was found. There were 9 cases in the oligospermia group. The age of the oligospermia patients was 22-30 years old, with an average age of 26±4 years old. The sperm counts in routine semen examinations were less than 20×10. 6 / mL. In the control group, there were 9 healthy male volunteers with a history of childbirth, aged 24 to 29 years old, with an average age of 26±3 years old, with normal semen.

Embodiment 2

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

[0040] (1) Screening of chip samples

[0041] Nine cases of azoospermia were selected from the collected clinical serum samples, aged from 23 to 29 years old, with an average age of 26±3 years old. No sperm was found in more than two routine semen examinations. There were 9 cases in the oligospermia group. The age of the oligospermia patients was 22-30 years old, with an average age of 26±4 years old. The sperm counts in routine semen examinations were less than 20×10. 6 / mL. In the control group, there were 9 healthy male volunteers with a history of childbirth, aged 24 to 29 years old, with an average age of 26±3 years old, with normal semen.

[0042] (2) Collection of serum samples

[0043] Collect 2ml of peripheral blood from the three groups to be tested, quickly transfer it to blood collection tube, let stand at room temperature for 15-30 minutes, centrifuge at 1900×g...

Embodiment 3

[0047] Example 3. Verification of real-time fluorescent quantitative PCR method for serum-specific miRNA in oligospermia and azoospermia

[0048] (1) Screening of verification samples

[0049] Twenty cases of azoospermia were selected from the collected clinical serum samples, aged from 23 to 35 years old, with an average age of 29±6 years old. No sperm was found in more than two routine semen examinations. There were 20 cases in the oligospermia group. The age of the patients with oligospermia was 22 to 34 years old, with an average age of 28±6 years old. The sperm counts in routine semen examinations were less than 20×10. 6 / mL. In the control group, there were 20 healthy male volunteers with a history of childbirth. They were 24-36 years old, with an average age of 30±6 years old, and had normal semen.

[0050] (2) Collection of serum samples

[0051] Collect 2ml of peripheral blood from the three groups to be tested, quickly transfer it to blood collection tube, let stand at room...

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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 type of single-stranded non-coding RNA composed of approximately 22 nucleotides. It mainly uses the direct shearing of messenger RNA or indirect inhibition of translation to regulate the expression of target genes at the post-transcriptional level. 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. Through the study of tissue miRNA expression profiles, it has been found that miRNAs have strong cell, tissue or disease specificity. These specifically expressed miNRAs are not only the basis of their functional studies, but also good disease markers. Recent studies have...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 曾骥孟李志明顾龙庄炫
Owner XIAMEN UNIV
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