Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Method for detecting neuraminidase influencing sperm functions

A technology of sialidase and detection method, which is applied in the field of medical detection, can solve the problems of inapplicability to clinical application, detection and reporting of sialidase activity, and failure to detect sialidase smoothly, so as to ensure accuracy and avoid dead sperm Effect

Active Publication Date: 2013-12-18
成都思瑞多医疗科技有限公司
View PDF2 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the literature only discloses the expression detection of sialidase in sperm, and does not further report the detection of sialidase activity; and the disclosed expression detection method cannot successfully detect sialidase in fresh sperm, and cannot Suitable for clinical application

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for detecting neuraminidase influencing sperm functions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The sialidase detection method affecting sperm function is characterized in that: the first step is to visually quantify the expression of sialidase in sperm by analyzing the fluorescence intensity of single sperm, or to visually observe the expression of sialidase in sperm by using fluorescence imaging under a microscope. If the sialidase is highly expressed in the sperm, proceed to the second step, using the in vitro capacitation fluid to capacious the sperm, and detect the sialidase activity with a fluorescent method.

Embodiment 2

[0046] The sialidase detection method affecting sperm function is characterized in that: the first step is to visually quantify the expression of sialidase in sperm by analyzing the fluorescence intensity of single sperm, or to visually observe the expression of sialidase in sperm by using fluorescence imaging under a microscope. If the sialidase is highly expressed in the sperm, proceed to the second step, using the in vitro capacitation fluid to capacious the sperm, and detect the sialidase activity with a fluorescent method.

[0047] The in vitro capacitation solution is HSA 5mg / ml HTF.

Embodiment 3

[0049] The implementation mode of this embodiment is basically the same as embodiment 1, on this basis:

[0050] Follow the steps below to detect the expression of sperm sialidase NEU1:

[0051] A Collect fresh liquefied semen samples, centrifuge to separate the sperm, and wash the sperm twice with PBS;

[0052] B Count the number of sperm and place on ice for 20 min;

[0053] C adopts 0.1% Triton X-100 pretreated the sperm, washed the sperm twice with PBS, and pretreated the sperm again with 3% PFA;

[0054] D uses 1% BSA-PBS solution to block;

[0055] E Incubate with 1ug / ul anti-human NEU1 antibody for 1 h, wash the sperm twice with PBS;

[0056] F At room temperature, the sperm were incubated with conventional fluorescent secondary antibodies at a ratio of 1:1000 for 1 h;

[0057] G Use flow cytometry or fluorescence microscope to make sperm cell smears, and observe the expression of sialidase in sperm.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a method for detecting neuraminidase influencing sperm functions. The method is a novel laboratory detecting method for assessing quality and functions of a sperm and can provide detection and clinical consultations for male infertility patients without clear clinical reasons for infertility. The method is characterized in that expressions of neuraminidase in the sperm are visually quantized through single sperm fluorescence strength analysis or are visually observed by means of fluorescence imaging under a microscope, and if the neuraminidase is highly expressed in the sperm, capacitation of the sperm is achieved through external capacitation liquid, and neuraminidase activity is detected in a fluorescence method.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a method for detecting sialidase that affects sperm function. Background technique [0002] With the continuous development of social industry, the total incidence of infertile couples increased significantly. In 2004, the statistics of the European Society of Reproduction: 25% of couples of childbearing age were unable to conceive within one year, 15% of them sought medical treatment, and 50% were infertility caused by male factors. The causes of male infertility are sexual dysfunction (1.7%), varicocele (12.3%), reproductive tract infection (6.6%), congenital dysplasia (2.1%) acquired disease (2.6%), Endocrine disorders (0.6%), immune factors (3.1%), and other abnormalities (3%), but up to 60% to 75% of patients cannot find the cause, which is called idiopathic male infertility, and they only show a few Abnormal sperm quality such as sperm, weak sperm or...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/573G01N33/531
Inventor 马芳许文明徐克惠岳焕勋蒋敏欧阳运薇林丽
Owner 成都思瑞多医疗科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com