Kit for evaluating semen quality and use method thereof

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

Active Publication Date: 2016-06-29
成都思瑞多医疗科技有限公司
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  • 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

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  • Kit for evaluating semen quality and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] A test kit for sperm quality assessment, the test kit includes cell lysate without enzyme activity inhibitors, detection buffer 1, protease inhibitor cocktail, reagent A solution, reagent B solution, bovine serum albumin, fluorescent Reagent, sialidase standard stock solution and detection buffer 2;

[0056] The detection buffer 1 is a phosphate buffer;

[0057] The reagent A liquid is: 1% BCA disodium salt, 2% anhydrous sodium carbonate, 0.16% sodium tartrate, 0.4% sodium hydroxide, 0.95% sodium bicarbonate, mixed to adjust the pH value to 11.2;

[0058] The reagent B liquid is: 4% copper sulfate;

[0059] The detection buffer 2 is a phosphate buffer containing 0.05 mmol / l sodium acetate and 0.25 μg / μl bovine serum albumin.

[0060] Adopt kit of the present invention to measure the sialidase activity of normal sperm (people) and sterile sperm (people) respectively, the result shows that the sialidase activity of normal sperm is obviously higher than that of sterile s...

Embodiment 2

[0062] A test kit for sperm quality assessment, the test kit includes cell lysate without enzyme activity inhibitors, detection buffer 1, protease inhibitor cocktail, reagent A solution, reagent B solution, bovine serum albumin, fluorescent Reagent, sialidase standard stock solution and detection buffer 2;

[0063] The detection buffer 1 is a phosphate buffer;

[0064] The reagent A liquid is: 1% BCA disodium salt, 2% anhydrous sodium carbonate, 0.16% sodium tartrate, 0.4% sodium hydroxide, 0.95% sodium bicarbonate, mixed to adjust the pH value to 11.3;

[0065] The reagent B liquid is: 4% copper sulfate;

[0066] The detection buffer 2 is a phosphate buffer containing 0.05 mmol / l sodium acetate and 0.25 μg / μl bovine serum albumin.

[0067] The sialidase standard stock solution is Clostridium perfringens sialidase with a concentration of 5 U / ml.

Embodiment 3

[0069] A test kit for sperm quality assessment, the test kit includes cell lysate without enzyme activity inhibitors, detection buffer 1, protease inhibitor cocktail, reagent A solution, reagent B solution, bovine serum albumin, fluorescent Reagent, sialidase standard stock solution and detection buffer 2;

[0070] The detection buffer 1 is BWW culture fluid;

[0071] The reagent A liquid is: 1% BCA disodium salt, 2% anhydrous sodium carbonate, 0.16% sodium tartrate, 0.4% sodium hydroxide, 0.95% sodium bicarbonate, mixed to adjust the pH value to 11.25;

[0072] The reagent B liquid is: 4% copper sulfate;

[0073] The detection buffer 2 is a phosphate buffer containing 0.05 mmol / l sodium acetate and 0.25 μg / μl bovine serum albumin.

[0074] The sialidase standard stock solution is Clostridium perfringens sialidase with a concentration of 5 U / ml.

[0075] The fluorescent reagent is sodium 2'-4-methylumbelliferyl-α-D-N-acetylneuraminic acid.

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Abstract

The invention provides a kit for evaluating semen quality and a use method thereof. The kit comprises a cell lysis buffer containing no enzyme activity inhibitor, a detecting buffer solution 1, a protease inhibitor cocktail, a reagent A solution, a reagent B solution, bovine serum albumin, a fluorescent reagent, a sialidase standard stock solution and a detecting buffer solution 2. The defecting buffer solution 1 is a phosphate buffer solution or BWW culture solution. The reagent A solution is prepared from 1% of BCA disodium salt, 2% of anhydrous sodium carbonate, 0.16% of sodium tartrate, 0.4% of sodium hydroxide and 0.95% of sodium bicarbonate, the raw materials are mixed, and the pH value is adjusted to be 11.2 to 11.3. The reagent B solution is prepared from 0.4% of copper sulfate. The detecting buffering solution 2 is a phosphate buffer solution containing 0.05 mol/l sodium acetate and 0.25 microgram/microlitre bovine serum albumin. The activity of sialidase in semen is detected, and diagnostic bases and clinical consulting are provided for male infertility patients with unclear clinical reasons.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a test kit for evaluating sperm quality and a use method thereof. 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 asthenoz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 马芳潘倩徐晨
Owner 成都思瑞多医疗科技有限公司
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