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Method for predicting whether sperms exist in testis of azoospermia patient or not according to seminal plasma mRNA

A technology for azoospermia and testis, applied in the field of medical testing, can solve the problem of inability to determine haploid sperm cells in patients with azoospermia, and achieve the effect of solving the problem of infertility and improving the success rate of sperm retrieval.

Pending Publication Date: 2020-08-11
广东省计划生育科学技术研究所
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Problems solved by technology

Currently, there is no way to determine the presence of haploid spermatids in the testes of azoospermic patients prior to surgical exploration

Method used

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  • Method for predicting whether sperms exist in testis of azoospermia patient or not according to seminal plasma mRNA
  • Method for predicting whether sperms exist in testis of azoospermia patient or not according to seminal plasma mRNA
  • Method for predicting whether sperms exist in testis of azoospermia patient or not according to seminal plasma mRNA

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Experimental program
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Embodiment 1

[0035] 1.1 Collection and processing of semen samples

[0036] 2ml of 30 normal semen and 42 azoospermic semen were collected and frozen in liquid nitrogen. Normal semen was obtained from the Human Sperm Bank of Guangdong Province. Azoospermia was defined as semen was taken every 4 to 5 weeks, and no sperm was found in microscopic examination after three semen centrifuges. Semen was centrifuged twice at low speed and the supernatant was used for RNA sequencing.

[0037] 1.2 Isolation and sequencing of RNA

[0038] The mirVana miRNA isolation kit can be used to separate large fragments and small fragments of RNA, and after separation, the quality, size distribution and quantity of RNA can be analyzed using Bioanalyer2100 (Agilent).

[0039] 1.3 Testicular multipoint biopsy

[0040] The patient was placed in the supine position, the surgical area was routinely disinfected, a sterile drape was spread, and internal spermatic nerve block anesthesia was performed with 1% lidocai...

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Abstract

The invention discloses a method for predicting whether sperms exist in the testis of an azoospermia patient or not according to seminal plasma mRNA. The method comprises the following steps of dividing seminal fluid specimens of the azoospermia patient into a testicular sperm group and a testicular azoospermia group by taking a testicular biopsy as a gold standard, determining the difference of exRNA expression profiles of two seminal fluid groups, carrying out comparison with an existing spermatogenic cell transcriptome database, and searching for differentially expressed exRNA capable of judging spermatogenic cell types, which is used for judging whether haploid sperm cells exist in the testis of the azoospermia patient or not. The clinical application of the scheme provided by the invention can greatly improve the sperm taking success rate of the azoospermia patient, and is also helpful for guiding the azoospermia patient to select an economical, painless and effective treatment method to solve the problem of infertility of the azoospermia patient.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to a method for predicting the presence or absence of sperm in the testis of azoospermia according to seminal plasma mRNA. Background technique [0002] About 10%-20% of male infertility patients are azoospermia, and azoospermia is divided into obstructive azoospermia (OA) and non-obstructive azoospermia (NOA). If there are sperms in the testicles of azoospermic patients, they can obtain sperms through surgical techniques such as testicular sperm extraction (TESE) and receive assisted reproductive technology so that they may obtain their own blood-related offspring. Almost 100% of OA patients can obtain them through TESE, etc. Sperm, while only about 50%-60% of NOA patients can obtain sperm. However, some azoospermia patients may still encounter the embarrassing situation of not being able to obtain sperm on the day of egg retrieval by the patient’s spouse. In NOA, this s...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/112
Inventor 闫威唐运革
Owner 广东省计划生育科学技术研究所
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