Detection of seminal plasma deoxycytidine and cytidine as diagnostic markers for idiopathic male infertility and its application

A technology for male infertility and deoxycytidine, which is applied in the fields of analytical chemistry and clinical medicine, can solve the problems of complex experimental steps and low sensitivity, and achieve the effects of improved sensitivity and specificity, high sensitivity and specificity, and easy detection

Active Publication Date: 2021-08-17
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Detection of seminal plasma deoxycytidine and cytidine as diagnostic markers for idiopathic male infertility and its application
  • Detection of seminal plasma deoxycytidine and cytidine as diagnostic markers for idiopathic male infertility and its application
  • Detection of seminal plasma deoxycytidine and cytidine as diagnostic markers for idiopathic male infertility and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] Example 1 Study object selection and grouping according to embodiment

[0106] This part of the study objects from Nanjing Medical University Hospital of first diagnosis of idiopathic cases of male infertility and reproductive health controls. Research and informed consent were approved by the ethics committee of the Medical University in Nanjing, meets the requirements of relevant laws and regulations. Cases and controls in the understanding of the content of signed informed consent. All subjects underwent a complete physical examination and completed a basic information including personal, lifestyle, occupational and environmental exposures, genetic risk factor questionnaire, sexual function and reproductive function, disease history and physical activity. The first stage 148 to meet the requirements included idiopathic male infertility cases and 148 healthy controls; 15 meet the requirements of the second stage idiopathic male infertility patients and 15 healthy controls...

Embodiment 2

[0138] Example 2 UPLC-MS metabolic sygental male infertility deoxytidine and cytosine screening

[0139] Pre-sample processing

[0140] 1.1. Take 10 μl of the semicake, 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 30s, precipitation protein.

[0141] 1.2. The centrifuge is centrifuged at 4 ° C for 15 min, and the supernatant was transferred to a 1.5 mL of the inlet EP tube, and the supernatant was concentrated in the centrifugal concentration dryer at room temperature.

[0142] 1.3. With 10 μl of ultra-pure water (reagent D), it is prepared.

[0143] 2. Instrument detection

[0144] 2.. Analysis Instrument: UPLC Ultimate 3000System (Dionex) High Performance Liquid Chromatograph; Q-EXACTIVE High Differentiation Mass Spectrometer.

[0145] 2.2. Liquid phase conditions:

[0146] 2.2.1 Liquid chromatography column is Hypersil Gold C18 column (100 mm × 2.1mm, particle size 1.9 μm, thermo ...

Embodiment 3

[0159] Example 3 Stability analysis of sequetal deoxytidine and cytanoside

[0160] The stability of the sequetal deoxycytidine and the cytosine level was evaluated using the method of Example 2 (the interval time was 2 weeks). The results show that the levels of deoxyphine and cytosine assay in the semers are stable ( figure 2 ), Have the characteristics of diagnostic / monitoring markers.

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Abstract

The invention belongs to the field of analytical chemistry and clinical medicine, and discloses the detection of seminal plasma deoxycytidine and cytidine as idiopathic male infertility diagnostic markers and its application. The marker is deoxycytidine and / or cytidine, has high sensitivity and specificity in diagnosing idiopathic male infertility, and can be used to prepare reagents for diagnosing or monitoring idiopathic male infertility.

Description

[0001] Invention [0002] The invention belongs to the analysis of analytical chemistry and clinical medicine, involving deoxyl cytosine and cytosine and its application based on UPLC-Q EXACTIVE MS detection in semers. Background technique [0003] In 1988, WHO first proposed the concept of reproductive health, that is, human and its individuals in the structure, functions and behaviors of the body, the physiology, function, and social harmonious health status of the structure, function and society in the process of reproduction throughout the life. At present, there are about 10-15% of childbearing age in the world with birth barriers. Due to the large population base, the number of infertile infertility in the newly married couple far exceeds millions, including male factors leading to less than 50%. Current research believes that genetic factors such as Y chromosome AZF gene region sequence changes, DAZ gene copy number change, sex hormone receptor gene sequence change, etc. Env...

Claims

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Application Information

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Patent Type & Authority Patents(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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