Application of serum miRNA biomarker

A biomarker and serum technology, applied in the field of biomarkers, can solve the problem that a single gene cannot fully explain the pathogenic mechanism, and achieve the effects of convenient source, accurate quantification, and convenient use

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

At present, most of the relevant studies explore the etiology of oligospermia and azoospermia from the perspective of a single gene, but the change of a single gene cannot fully explain their pathogenic mechanism

Method used

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  • Application of serum miRNA biomarker
  • Application of serum miRNA biomarker
  • Application of serum miRNA biomarker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, collection of clinical samples and arrangement of clinical data

[0045] According to the standards of the World Health Organization (“Laboratory Test Manual for Human Semen and Sperm-Cervical Mucus Interaction [M]” (4th Edition), edited by the World Health Organization, translated by Gu Qun, etc., Beijing: People’s Medical Publishing House, 2001.5.1 ) determined as azoospermia or severe oligospermia. The semen examination should be analyzed at least twice (including centrifuge thick section analysis). All the subjects were married for 2 to 13 years, the woman's examination was normal, they lived together for more than 2 years after marriage, and no contraceptive measures were taken. After strict screening, the infertility caused by various infection sequelae, vas deferens obstruction and hypothalamus-pituitary disease were excluded. No endocrine therapy was performed. Cytogenetic analysis showed a normal karyotype.

Embodiment 2

[0046] Example 2. Preliminary screening of serum-specific miRNA expression chip for oligospermia and azoospermia

[0047] (1) Screening of chip samples

[0048]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.

[0049] (2) Collection of serum samples

[0050] Collect 2ml of peripheral blood from three groups of subjects, quickly transfer it to 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 a centrifuge t...

Embodiment 3

[0054] Example 3, Real-time fluorescence quantitative PCR method verification of oligospermia and azoospermia serum-specific miRNA

[0055] (1) Screening of verification samples

[0056] 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.

[0057] (2) Collection of serum samples

[0058] Collect 2ml of peripheral blood from three groups of subjects, quickly transfer it to 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 supe...

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Abstract

The invention provides application of a serum miRNA biomarker, and relates to biomarkers. The serum miRNA biomarker comprises at least one of miR-34c, miR-1275, miR-375, miR-410 and miR-758. The serum miRNA biomarker can be applied to preparation of a reagent for detecting oligospermia and azoospermia. A serum miRNA biomarker composition comprises at least one of the miRNA and sequences of reverse transcription primers, forward primers and reverse primers of each miRNA, and can be applied to preparation of a reagent for detecting oligospermia and azoospermia. The determined close correlation between the miRNA combination and indexes of oligospermia and azoospermia cases can be used as the biomarker to screen and diagnose patients with oligospermia and azoospermia, and an efficient basis is provided to clinical individual intervention treatment.

Description

technical field [0001] The present invention relates to biomarkers, in particular to the application of a serum miRNA biomarker. technical background [0002] Infertility is a global medical problem. According to the statistics of the World Health Organization, about 5% to 10% of couples of childbearing age in the world have infertility problems, and male infertility accounts for about half of them. 90% of male infertility is caused by spermatogenesis disorder, clinically manifested as azoospermia or oligospermia. Oligospermia means that there are less than 20 million sperm per milliliter of semen. Because the number of sperm is low, the chance of meeting the egg is less, resulting in male infertility. Azoospermia accounts for about 15% to 20% of male infertility patients. Azoospermia refers to multiple semen examinations (generally more than 3 times) but no sperm was found. If necessary, patients with azoospermia can undergo testicular biopsy, spermography, endocrine hor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/68
Inventor 曾骥孟李志明顾龙庄炫
Owner XIAMEN UNIV
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