Method for evaluating semen quality

A technology of sperm quality and sperm, which is applied in the field of sperm quality assessment, can solve the problems of high price, complicated technical process, high requirements for experimental equipment and operators' skills and experience, and achieve the goal of reducing negative impact, simple operation method, and few types of consumables Effect

Active Publication Date: 2016-06-15
成都思瑞迪医疗科技有限公司
View PDF4 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the sialidase activity of the sperm itself is not necessarily related to the enzyme activity of the capacitation fluid, this method cannot detect the sialidase contained in the sperm itself, and cannot evaluate the quality of the sperm itself
At the same time, there are still the following disadvantages in the method: 1) The technical process is complicated; 2) It requires high skills and experience for experimental equipment and operators; 3) It is not suitable for clinical laboratories and routine technicians; 4) Use abcam company kits are expensive

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 evaluating semen quality

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] A method for sperm quality assessment, the specific steps are as follows:

[0051] A. Sperm washing

[0052] Collect fresh liquefied semen samples, wash the sperm with phosphate buffer, fully remove the remaining seminal plasma components, and then centrifuge to separate the sperm;

[0053] B. Extraction of sperm protein

[0054] Add cell lysate and protease inhibitor cocktail to the sperm separated in step A, mix well, and fully lyse when there is no obvious cell precipitation, centrifuge at 4°C at high speed, and the supernatant is the protein extract;

[0055] C. Determination of protein concentration of sperm protein extract

[0056] (1) Use the cell lysate to prepare the protein standard stock solution of bovine serum albumin, and then use the lysate to dilute the protein standard stock solution into the protein standard solution according to the concentration gradient of 2 / 1.0 / 0.5 / 0.25 / 0.125 / 0 mg / ml, and then use it to detect Add each protein standard solution ...

Embodiment 2

[0063] A method for sperm quality assessment, the specific steps are as follows:

[0064] A. Sperm washing

[0065] Collect fresh liquefied semen samples, wash the sperm with phosphate buffer, fully remove the remaining seminal plasma components, and then centrifuge to separate the sperm;

[0066] B. Extraction of sperm protein

[0067] Add cell lysate and protease inhibitor cocktail to the sperm separated in step A, mix well, and fully lyse when there is no obvious cell precipitation, centrifuge at 4°C at high speed, and the supernatant is the protein extract;

[0068] C. Determination of protein concentration of sperm protein extract

[0069] (1) Use the cell lysate to prepare the protein standard stock solution of bovine serum albumin, and then use the lysate to dilute the protein standard stock solution into the protein standard solution according to the concentration gradient of 2 / 1.0 / 0.5 / 0.25 / 0.125 / 0 mg / ml, and then use it to detect Add each protein standard solution ...

Embodiment 3

[0076] A method for sperm quality assessment, the specific steps are as follows:

[0077] A. Sperm washing

[0078] Collect fresh liquefied semen samples, wash the sperm with BWW culture medium, fully remove the remaining seminal plasma components, and then centrifuge to separate the sperm;

[0079] B. Extraction of sperm protein

[0080] Add cell lysate and protease inhibitor cocktail to the sperm separated in step A, mix well, and fully lyse when there is no obvious cell precipitation, centrifuge at 4°C at high speed, and the supernatant is the protein extract;

[0081] C. Determination of protein concentration of sperm protein extract

[0082] (1) Use the cell lysate to prepare the protein standard stock solution of bovine serum albumin, and then use the lysate to dilute the protein standard stock solution into the protein standard solution according to the concentration gradient of 2 / 1.0 / 0.5 / 0.25 / 0.125 / 0 mg / ml, and then use it to detect Add each protein standard solutio...

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 evaluating semen quality. The method comprises the following specific steps: 1, collecting a fresh liquefied seminal fluid sample, washing semen with a phosphate buffer solution or a BWW culture solution, removing residual seminal plasma constituents completely, and then conducting centrifugal separation on the semen; 2, adding cell lysis buffer and a protease inhibiting agent cocktail to the semen obtained through separation in step 1, mixing the mixture to be uniform, and after no obvious cell sedimentation is found, in other words, complete splitting decomposition is achieved, conducting high speed centrifugation at the temperature of 4 DEG C, wherein supernatant is protein extract; 3, measuring the protein concentration of the protein extract; 4, measuring sialidase activity of the semen, and finally obtaining the sialidase activity value / protein concentration value, so that the process is ended. According to the method, through detection of activity of sialidase in the semen, the diagnosis basis and clinical reference are provided for a patient suffering from male infertility with the clinical reasons unclear.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a method for evaluating sperm quality. 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 oligospermia, Abnormal sperm quality such as asthenozoospermia or teratozoosperm...

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): C12Q1/34
Inventor马芳马学李福平
Owner成都思瑞迪医疗科技有限公司