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CCL8 protein as biomarker of non-obstructive azoospermia and application of CCL8 protein

A biomarker, azoospermia technology, applied in the field of biomarkers of non-obstructive azoospermia, 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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  • CCL8 protein as biomarker of non-obstructive azoospermia and application of CCL8 protein
  • CCL8 protein as biomarker of non-obstructive azoospermia and application of CCL8 protein

Examples

Experimental program
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Embodiment 1

[0043] This embodiment provides a diagnostic kit for non-obstructive azoospermia, which contains an antibody to CCL8 protein (R&D Systems, MAB281-100), a secondary antibody to HRP (R&D Systems, HAF008), and a CCL8 protein standard ( R&DSystems, 281-CP-010 / CF) 96-well plate, diluent, chromogenic reagent and stop solution, wherein the diluent is PBS, chromogenic reagent (substrate chromogenic solution A: take 13.6 g of sodium acetate, 1.6 g of citric acid g and 30% hydrogen peroxide 0.3mL, add distilled water to 500mL; substrate color development 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 solution 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 CCL8 protein antibody, dilute the antibody to a concentration of 500ng / mL, and add the diluted antibody to the 96-well plate...

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 CCL8 protein in each semen sample was detected using the kit in Example 1, including the following steps:

[0048] (1) The 80ng / mL CCL8 protein standard was diluted by comparison to obtain 80ng / mL, 40ng / mL, 20ng / mL, 10ng / mL, 5ng / mL and 0ng / mL standard solution for making a standard curve;

[0049] (2) The new type of semen sample obtained by masturbation of the patient 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 washin...

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Abstract

The invention discloses a CCL8 protein serving as a biomarker of non-obstructive azoospermia and application of the CCL8 protein. The biomarker comprises a CCL8 protein. The invention finds that the concentration of the CCL8 protein in the seminal fluid of the NOA patient is obviously lower than that of a normal male for the first time, and proposes that the CCL8 protein can be used as a potential early warning molecule for evaluating the spermatogenic dysfunction of the NOA patient, so that expansion and diagnosis of NOA indexes are facilitated, the diagnosis range is expanded, and the CCL8 protein has important significance in the field of NOA diagnosis.

Description

technical field [0001] The invention belongs to the technical field of reproductive medicine, and relates to CCL8 protein as a biomarker of 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-obst...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/689G01N33/6863G01N33/54306G01N2800/367G01N2333/521
Inventor 赵亮宇汤育新陈玉琢叶昆彭经璇楚溪
Owner THE FIFTH AFFILIATED HOSPITAL SUN YAT SEN UNIV
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