Quantitative detection method and kit of sperm acrosin activity

A detection method and quantitative detection technology, which is applied in the direction of material analysis by observing the influence of chemical indicators, and analysis by making materials undergo chemical reactions, etc., which can solve the problems of difficult to achieve synchronous reaction, cumbersome operation, and influence on the accuracy of detection results And other issues

Active Publication Date: 2012-02-15
BRED LIFE SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the Kennedy method is cumbersome to operate: the sperm must be centrifuged again after the reaction is completed, and each sample needs to be set up for measurement and control wells. It is difficult to achieve simultaneous reactions during batch detection, which affects the accuracy of the detection results. The reaction time is longer (3 hours) )and many more

Method used

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  • Quantitative detection method and kit of sperm acrosin activity
  • Quantitative detection method and kit of sperm acrosin activity
  • Quantitative detection method and kit of sperm acrosin activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] One, prepare the sperm acrosome activity quantitative detection kit of the present invention

[0083] 1. Install the reaction device:

[0084] 1) Weigh 13.61 grams of potassium dihydrogen phosphate and dissolve in 1L of purified water, weigh 35.814 grams of disodium hydrogen phosphate 12 water and dissolve in 1L of purified water, and mix the two solutions until the pH value is 7 under the monitoring of a pH meter. The mixed solution is pH7, 0.1mol / L phosphate buffer solution. (pH7, 0.1mol / L phosphate buffer)

[0085] 2) Weigh 0.5 g of L-aminocaproic acid-globulin polymer, put it in a beaker, add pH 7, 0.1mol / L phosphate buffer and stir to dissolve, then use pH 7, 0.1mol / L phosphate buffer to volume to 1L, mix well and save. (The concentration of optically active aminocaproic acid polymer is 0.05%, which means that every 100 milliliters of buffer solution contains 0.05 grams of L-aminocaproic acid-globulin polymer)

[0086] 3) Select a polytetrafluoroethylene membra...

Embodiment 2

[0155] One, prepare the sperm acrosome activity quantitative detection kit of the present invention

[0156] 1. Install the reaction device:

[0157] 1) Weigh 1.361 grams of potassium dihydrogen phosphate and dissolve in 1L of purified water, weigh 3.5814 grams of disodium hydrogen phosphate 12 water and dissolve in 1L of purified water, and mix the two solutions until the pH value is 5 under the monitoring of a pH meter. The mixed solution is pH5, 0.01mol / L phosphate buffer solution. (pH5, 0.01mol / L phosphate buffer)

[0158] 2) Weigh 100.00 grams of L-aminocaproic acid-agarose polymer, put it in a beaker, add pH 5, 0.01mol / L phosphate buffer, stir to dissolve, then use pH 5, 0.01mol / L phosphate buffer to make up the volume to 1L, mix well and save. (The concentration of optically active aminocaproic acid polymer is 10%, which means that every 100 milliliters of buffer contains 10 grams of L-aminocaproic acid-globulin polymer)

[0159] 3) Take a nitrocellulose membrane (C...

Embodiment 3

[0228] One, prepare the sperm acrosome activity quantitative detection kit of the present invention

[0229] 1. Install the reaction device:

[0230] 1) Weigh 136.1 grams of potassium dihydrogen phosphate and dissolve in 1L of purified water, weigh 358.14 grams of disodium hydrogen phosphate 12 water and dissolve in 1L of purified water, and mix the two solutions until the pH value is 9 under the monitoring of a pH meter. The mixed solution is pH9, 1mol / L phosphate buffer solution. (pH9, 1mol / L phosphate buffer)

[0231] 2) Weigh 20 grams of L-aminocaproic acid polymer complex, put it in a beaker, add pH 9, 1mol / L phosphate buffer and stir to dissolve, then dilute to 1L with pH 9, 1mol / L phosphate buffer , and store after mixing. (The concentration of optically active aminocaproic acid polymer is 2.0%, which means that every 100 milliliters of buffer contains 2.0 grams of L-aminocaproic acid-globulin polymer)

[0232] 3) Take a polypropylene fiber membrane (PP) with a pore...

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Abstract

The invention relates to a solid phase filter membrane detection method of sperm acrosin activity. The method comprises steps that: sperms are absorbed on the surface of the solid phase filter membrane; liquid and impurities other than sperms are removed from the surface of the solid phase filter membrane by using a centrifugation method or a dumping method; an acrosin activity detection reactionis started, wherein the sperms absorbed on the surface of the solid phase filter membrane contact a substrate solution, such that the acrosin activity detection reaction is started; the reaction is carried out for 30 to 120 minutes under a temperature of 4 to 40 DEG C; the substrate is separated from the sperms on the solid phase, or a quality control stop buffer is added in; reaction products are processed through colorimetry; and acrosin activity is obtained by calculating the absorbance values obtained through colorimetry. The invention also relates to a quantitative detection kit for sperm acrosin activity. According to the invention, a special-purposed reaction apparatus is adopted, the operation is simple, the result is accurate, and the detection means is flexible. The method and the kit are suitable for batch detections, and are easy to popularize.

Description

technical field [0001] The invention relates to a method and a device for in vitro detection, in particular to a method and a kit suitable for detecting sperm acrosome activity. Background technique [0002] Acrosinase exists between the inner acrosome membrane of the sperm head and the equatorial membrane. When the sperm combines with the oocyte, the acrosome reaction occurs at the head of the sperm, the acrosome outer membrane ruptures, and the acrosome enzyme is released. Acrosin is an important proteolytic enzyme in the fertilization process, which can dissolve the corona radiata and zona pellucida around the oocyte, so that the sperm can pass through the zona pellucida and fuse with the egg cell to complete fertilization. Reduced sperm acrosome enzyme activity can affect the penetration of sperm through the zona pellucida of the oocyte, resulting in infertility. In addition to the quality of the sperm itself, severe reproductive system infections can also reduce the a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
Inventor 胡家纯钟彩颜王希上张翔程锦军庄学敏
Owner BRED LIFE SCI TECH
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