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Molecular marker related to azoospermia in semen as well as detection method and application of molecular marker

A technique for molecular markers and azoospermia, which is applied in the fields of analytical chemistry and clinical medicine, can solve problems that have not received corresponding attention, and achieve the effects of preventing disease progression, accurate quantification, and delaying disease progression

Pending Publication Date: 2022-04-12
苏州南医大创新中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Seminal plasma, as the liquid component of semen, can well reflect the quality of semen, and has the advantages of being non-invasive and easy to collect. However, the application of metabolomics analysis of seminal plasma metabolic small molecules in the diagnosis and monitoring of idiopathic male infertility Has not yet received the corresponding attention

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  • Molecular marker related to azoospermia in semen as well as detection method and application of molecular marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Embodiment 1 Research Object Selection and Grouping Basis

[0074] The subjects of this part of the study were first-diagnosed azoospermia cases and normal controls with normal semen parameters from the Affiliated Hospital of Nanjing Medical University and the Reproductive Medicine Center of the Sixth Affiliated Hospital of Sun Yat-sen University. The research content and the informed consent form were approved by the Ethics Committee of Nanjing Medical University and complied with the requirements of relevant laws and regulations. The cases and controls signed the informed consent form after understanding the contents. All the research subjects underwent a complete physical examination and completed a report that included personal basic information, living habits, occupational and environmental exposures, genetic risk factors, sexual function and reproduction. Questionnaires on function, disease history, and physical activity. Specific sample classification criteria a...

Embodiment 2

[0089] Example 2 UPLC-MS metabolomics screening of azoospermia 1,3-dihydroxypropan-2-yl 13-methyltetradecanoic acid

[0090] 1. Sample pretreatment

[0091] 1.1. Take 10 μL of semen, add 10 μL of internal standard A, add 10 μL of internal standard B, add 10 μL of internal standard C, add 40 μL of methanol (reagent A), vortex for 30 seconds, and precipitate protein.

[0092] 1.2. Centrifuge at 16000g at 4°C for 15min in a centrifuge, transfer the supernatant to a 1.5mL imported EP tube, and concentrate the supernatant to dryness in a centrifugal concentrator at room temperature.

[0093] 1.3. Reconstitute with 10 μL of ultrapure water (reagent D) and wait for analysis.

[0094] 2. Instrument testing

[0095] 2.1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.

[0096] 2.2. Liquid phase conditions:

[0097] 2.2.1 The liquid chromatographic column is a Hypersil GOLD C18 chromatogr...

Embodiment 3

[0108] Example 3 Diagnosis of azoospermia by 1,3-dihydroxypropan-2-yl 13-methyltetradecanoic acid

[0109] According to the above-mentioned UPLC-Q exactive MS metabolomics method, the inventors drew the ROC curve by detecting 1,3-dihydroxypropan-2-yl-13-methyltetradecanoic acid in the semen samples of 62 cases and 84 controls And evaluate the sensitivity and specificity of prediction, and then evaluate the ability of detecting the level of this metabolic small molecule in semen to evaluate azoospermia.

[0110] See figure 1 , the sensitivity of 1,3-dihydroxypropan-2-yl 13-methyltetradecanoic acid is 96.3%, the specificity is 91.4%, and the area under the ROC curve is 0.965, so 1,3-dihydroxypropan-2-yl 13-methyltetradecanoic acid has a good ability to diagnose azoospermia.

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Abstract

The invention relates to a molecular marker related to azoospermia in semen as well as a detection method and application of the molecular marker, and belongs to the technical field of analytical chemistry and clinical medicine. Metabolic small molecules in normal control and azoospermia seminal fluid are compared by adopting UPLC-Qexactive MS (Ultra Performance Liquid Chromatography-Quantitative Exactive MS), 1, 3-dihydroxypropyl-2-yl 13-methylmyristic acid which can be used for evaluating whether the seminal fluid suffers from azoospermia or not and has a diagnostic value is found to exist in the seminal fluid, and the application of the UPLC-Qexactive MS for detecting the 1, 3-dihydroxypropyl-2-yl 13-methylmyristic acid is found to be used for evaluating whether the seminal fluid suffers from azoospermia or not. And the azoospermia diagnosis and monitoring kit convenient for clinical application is developed. Metabolic small molecules are novel biomarkers, are highly associated with disease outcomes, and are stable, non-invasive, easy to detect and accurate in quantification. The 1, 3-dihydroxypropyl-2-yl 13-methylmyristic acid provided by the invention can be used as a diagnostic marker of azoospermia, provides a basis for further examination of clinicians, provides support for quickly and accurately mastering disease states of patients and timely taking prevention and treatment schemes, and delays and prevents disease progression.

Description

technical field [0001] The invention relates to a molecular marker related to azoospermia in semen, a detection method and application thereof, and belongs to the technical fields of analytical chemistry and clinical medicine. Background technique [0002] According to the statistics of the World Health Organization (WHO), about 15% of couples of childbearing age in the world suffer from infertility. Due to the large population base in our country, the number of infertility patients among newly married couples is far more than one million, and the infertility caused by male factors is higher than 50%. Infertility is a common problem among couples worldwide, an estimated 4-17% of couples visit a doctor to correct their infertility, but it is generally believed that more infertile couples do not go diagnosis and treatment. Surveys show that the male factor is the main or only factor accounting for about 20-50% of the causes of infertility. The incidence of azoospermia accou...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72G01N33/50C07C59/105
Inventor 沙家豪袁艳陈敏健杨晓玉王嘉琛姚莹
Owner 苏州南医大创新中心
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