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Detection of seminal plasma mannose-6-phosphoric acid and neopterin as diagnostic marker for idiopathic male sterility and application thereof

A male infertility, idiopathic technology, applied in the field of analytical chemistry and clinical medicine, can solve the problems of complex experimental steps and low sensitivity, and achieve the goal of improving sensitivity and specificity, high sensitivity and specificity, and preventing disease progression. Effect

Active Publication Date: 2020-12-25
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 mannose-6-phosphoric acid and neopterin as diagnostic marker for idiopathic male sterility and application thereof
  • Detection of seminal plasma mannose-6-phosphoric acid and neopterin as diagnostic marker for idiopathic male sterility and application thereof
  • Detection of seminal plasma mannose-6-phosphoric acid and neopterin as diagnostic marker for idiopathic male sterility and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Embodiment 1 Research Object Selection and Grouping Basis

[0084] 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

[0115] Example 2 UPLC-MS metabolomics screening of idiopathic male infertility mannose-6-phosphate and neopterin

[0116] 1. Sample pretreatment

[0117] 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.

[0118]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.

[0119] 1.3. Reconstitute with 10 μL of ultrapure water (reagent D) and wait for analysis.

[0120] 2. Instrument testing

[0121] 2.1. Analytical instruments: UPLC Ultimate 3000system (Dionex) high-performance liquid chromatography; Q-Exactive high-resolution mass spectrometer.

[0122] 2.2. Liquid phase conditions:

[0123] 2.2.1 The liquid chromatographic column is a Hypersil GOLD C18 chrom...

Embodiment 3

[0136] Example 3 Stability analysis of seminal plasma mannose-6-phosphate and neopterin

[0137] The method of Example 2 was used to evaluate the stability of seminal plasma mannose-6-phosphate and neopterin levels (the interval was 2 weeks). The results showed that the measured levels of mannose-6-phosphate and neopterin 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 provides UPLC-Q exactive MS detection of mannose-6-phosphoric acid and neopterin related to idiopathic male sterility in seminal plasma and application thereof. According to the invention, the UPLC-Q exactive MS is adopted to compare metabolic small molecules in normal control and idiopathic male sterility seminal plasma, and the fact that the seminal plasma contains a combination of mannose-6-phosphoric acid and neopterin which can be used for evaluating whether a patient suffers from idiopathic male sterility and has diagnostic value is found, and the application of the UPLC-Q exactive MS for detecting mannose-6-phosphoric acid and neopterin is found, so that an idiopathic male sterility diagnosis and monitoring kit which can be conveniently applied clinically is developed.

Description

[0001] field of invention [0002] The invention belongs to the field of analytical chemistry and clinical medicine, and relates to detection of mannose-6-phosphate and neopterin in seminal plasma related to idiopathic male infertility based on UPLC-Q active MS and application thereof. Background technique [0003] In 1988, the World Health Organization (WHO) first proposed the concept of reproductive health, that is, the complete and harmonious physiological, psychological and social conditions of human beings and their individuals in the structure, function and behavior of reproductive-related organisms throughout life. health status. 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 research suggests that genetic factors and env...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/06G01N30/88G01N2030/045G01N2030/8813
Inventor 陈敏健王洁黄芮段家伟胡艳辉管昱盛张晓玲张婷陈道桢王玉邦王心如夏彦恺
Owner NANJING MEDICAL UNIV
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