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Detection of seminal plasma mannose-6-phosphate and neopterin as diagnostic markers for idiopathic male infertility and its application

A male infertility and marker technology, applied in the fields of analytical chemistry and clinical medicine, can solve the problems of complex experimental steps and low sensitivity, and achieve the effect of improving sensitivity and specificity, high sensitivity, and accurate quantification.

Active Publication Date: 2021-07-20
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-phosphate and neopterin as diagnostic markers for idiopathic male infertility and its application
  • Detection of seminal plasma mannose-6-phosphate and neopterin as diagnostic markers for idiopathic male infertility and its application
  • Detection of seminal plasma mannose-6-phosphate and neopterin as diagnostic markers for idiopathic male infertility and its application

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 contents. 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 inclu...

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 fields of analytical chemistry and clinical medicine, and provides a method for detecting mannose-6-phosphate and neopterin in seminal plasma related to idiopathic male infertility by UPLC-Q active MS and an application thereof. The present invention compares the metabolic small molecules in the seminal plasma of the normal control and idiopathic male infertility by using UPLC-Q exact MS, and finds that there are small molecules in the seminal plasma that can be used to evaluate whether there is idiopathic male infertility, and have diagnostic value. The combination of mannose-6-phosphate and neopterin, as well as the application of UPLC-Q exact MS for the detection of mannose-6-phosphate and neopterin, have developed an idiopathic male infertility diagnosis and monitoring that can be easily applied clinically Reagent test kit.

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 Patents(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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