Diagnostic marker of idiopathic male infertility by detecting deoxycytidine and cytidine in seminal plasma and application thereof

A technology for male infertility and deoxycytidine, applied in the fields of analytical chemistry and clinical medicine, can solve the problems of complex experimental steps and low sensitivity

Active Publication Date: 2019-01-11
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

NMR technology is characterized by no damage to the components to be measured, simple sample pretreatment, but low sensitivity; gas chromatography-mass spectrometry has good sensitivity and reproducibility, but derivatization methods are generally used for sample pretreatment , making the experimental steps more complicated

Method used

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  • Diagnostic marker of idiopathic male infertility by detecting deoxycytidine and cytidine in seminal plasma and application thereof
  • Diagnostic marker of idiopathic male infertility by detecting deoxycytidine and cytidine in seminal plasma and application thereof
  • Diagnostic marker of idiopathic male infertility by detecting deoxycytidine and cytidine in seminal plasma and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Embodiment 1 Research Object Selection and Grouping Basis

[0105] The subjects of this part of the study are from the first-diagnosed idiopathic male infertility cases and healthy childbearing controls in the Affiliated Hospital of Nanjing Medical 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. Cases and controls signed the informed consent after understanding the content. All subjects underwent a complete physical examination and completed a questionnaire including basic personal information, living habits, occupational and environmental exposures, genetic risk factors, sexual and reproductive function, disease history, and physical activity. 148 eligible idiopathic male infertility cases and 148 healthy controls were included in the first stage; 15 idiopathic male infertility cases and 15 healthy controls were includ...

Embodiment 2

[0137] Example 2 UPLC-MS metabolomics screening of deoxycytidine and cytidine in idiopathic male infertility

[0138] 1. Sample pretreatment

[0139] 1.1. Take 10 μL of seminal plasma, 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.

[0140] 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.

[0141] 1.3. Reconstitute with 10 μL of ultrapure water (reagent D) and wait for analysis.

[0142] 2. Instrument testing

[0143] 2.1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.

[0144] 2.2. Liquid phase conditions:

[0145] 2.2.1 The liquid chromatographic column is a Hypersil GOLD C18 chromato...

Embodiment 3

[0158] Example 3 Stability analysis of seminal plasma deoxycytidine and cytidine

[0159] The method of Example 2 was used to evaluate the stability of seminal plasma deoxycytidine and cytidine levels (the interval was 2 weeks). The results showed that the measured levels of deoxycytidine and cytidine in seminal plasma were stable ( figure 2 ), with properties as diagnostic / monitoring markers.

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Abstract

The invention belongs to the field of analytical chemistry and clinical medicine, and discloses a diagnostic marker of idiopathic male infertility by detecting deoxycytidine and cytidine in seminal plasma and an application thereof. The marker is deoxycytidine and/or cytidine, has high sensitivity and specificity in the diagnosis of the idiopathic male infertility, and can be used for preparing adiagnosis or monitoring reagent for idiopathic male infertility.

Description

field of invention [0001] The invention belongs to the field of analytical chemistry and clinical medicine, and relates to detection of deoxycytidine and cytidine in seminal plasma related to idiopathic male infertility based on UPLC-Q active MS and application thereof. Background technique [0002] In 1988, the WHO first proposed the concept of reproductive health, which refers to the complete and harmonious state of physiology, psychology and society of human beings and their individuals in the structure, function and behavior of reproductive-related organisms throughout their life. Currently, about 10-15% of couples of reproductive age worldwide suffer from fertility disorders. Due to the large population base in my country, the number of infertility patients among newlyweds is far more than one million, and the infertility caused by male factors is higher than 50%. Current studies suggest that genetic factors such as sequence changes in the AZF gene region of the Y chro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72
Inventor 陈敏健丁兴旺庄婷钰管昱盛徐菠周昆胡艳辉冒志磊霍薇薇
Owner NANJING MEDICAL UNIV
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