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Biomarker related to non-obstructive azoospermia and application thereof

A biomarker, azoospermia technology, applied in the field of reproductive medicine, can solve problems such as difficult quantitative detection

Pending Publication Date: 2022-02-18
THE FIFTH AFFILIATED HOSPITAL SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, hsa_circMAN1A2 can be used as a circRNA marker for the diagnosis of non-obstructive azoospermia; CN112176052A discloses a plasma exosomal tsRNA marker and its application related to the diagnosis of non-obstructive azoospermia. The marker is tRF-Gly -GCC-002, which is differentially expressed in the plasma of patients with non-obstructive azoospermia and obstructive azoospermia, can better diagnose non-obstructive azoospermia. It can be seen that a variety of diagnostic NOA indicators have been developed, but These indicators are mainly concentrated in non-coding RNA, its quantitative detection is relatively difficult, and there are still some patients who cannot use these indicators to make a correct diagnosis

Method used

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  • Biomarker related to non-obstructive azoospermia and application thereof
  • Biomarker related to non-obstructive azoospermia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] This embodiment provides a diagnostic kit for non-obstructive azoospermia, the kit contains SHISA5 protein antibody (Novus Biologicals, H00051246-M03), SHISA5 protein standard (Novus Biologicals, NBP1-83190PEP) 96-well plate , diluent, chromogenic reagent and stop solution, wherein the diluent is PBS, and the chromogenic reagent is (substrate color development solution A: take 13.6g of sodium acetate, 1.6g of citric acid and 0.3mL of 30% hydrogen peroxide, add distilled water to 500mL ; Substrate Chromogenic Solution B: Take 0.2g disodium edetate, 0.95g citric acid, 50mL glycerin and 0.15g TMB (dissolved in 3mL DMSO), add distilled water to 500mL, ready to use), stop The liquid is sulfuric acid.

[0044]In addition, the 96-well plate needs to be coated. The coating method is: use phosphate buffer (pH 7.2) to dilute the SHISA5 protein antibody, dilute the antibody to a concentration of 500ng / mL, and add the diluted antibody to the 96-well plate. Place in a refrigerator ...

Embodiment 2

[0046] In this example, the kit in Example 1 is used to detect the semen sample.

[0047] The semen of 10 NOA patients and the semen of 10 normal men were respectively taken, and the concentration of SHISA5 protein in each semen sample was detected using the kit in Example 1, including the following steps:

[0048] (1) Dilute the 20ng / mL SHISA5 protein standard by comparison to obtain 20ng / mL, 10ng / mL, 5ng / mL, 2.5ng / mL, 1.25ng / mL and 0ng / mL standard solution for making a standard curve ;

[0049] (2) The new type of semen sample obtained by the patient's masturbation was placed in a 37°C water bath for 30 minutes for liquefaction;

[0050] (3) After the semen is completely liquefied, centrifuge at 3000g / 5min, and take the supernatant seminal plasma;

[0051] (4) Take 100mL of seminal plasma and standard products of various concentrations, place them in a 96-well plate coated with antibodies, and incubate at 37°C for 30min;

[0052] (5) After incubation, wash 3 times with wa...

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Abstract

The invention discloses a biomarker related to non-obstructive azoospermia and an application of the biomarker. The biomarker comprises an SHISA5 protein. According to the present invention, the concentration of the Wnt receptor inhibition protein SHISA5 in the NOA patient semen is significantly lower than the concentration of the Wnt receptor inhibition protein SHISA5 in the normal male semen for the first time, and the SHISA5 protein can be adopted as the potential early warning molecule for evaluating the spermatogenic dysfunction of the NOA patient, such that the NOA diagnosis index can be easily expanded so as to expand the diagnosis range, and the important significance is provided for the NOA diagnosis field.

Description

technical field [0001] The invention belongs to the technical field of reproductive medicine, and relates to a biomarker related to non-obstructive azoospermia and its application. Background technique [0002] The incidence of azoospermia accounts for 10%-20% of male infertility. According to the presence or absence of sperm in the testes, it can be divided into two categories: one is obstructive azoospermia (OA) with normal spermatogenesis in the testis, and the other is non-obstructive azoospermia (non-obstructive azoospermia) with impaired testicular spermatogenesis. -obstructive azoospermia, NOA). An accurate diagnosis of non-obstructive azoospermia can provide valuable guidance to clinicians. [0003] Testicular volume, serum follicle-stimulating hormone (FSH) level, etc. can assist in the diagnosis of NOA patients. It has also been reported that RNA is used as a marker of NOA. For example, CN113215246A discloses a circRNA marker for the diagnosis of non-obstructive ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543C07K14/47
CPCG01N33/689G01N33/54306C07K14/4703G01N2333/4704G01N2800/367
Inventor 赵亮宇汤育新戴英波叶昆何侃成谢能庆
Owner THE FIFTH AFFILIATED HOSPITAL SUN YAT SEN UNIV
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