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A sperm chromatin structure detection kit and application thereof

A technology for structure detection and chromatin, which is applied in the field of medicine, can solve problems such as unanalyzable, unanalyzable fragmentation, proportion of immature sperm, and many product components, so as to simplify detection steps and operation time, and facilitate clinical and scientific research work the effect of

Active Publication Date: 2019-01-01
ZHEJIANG BOZHEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The detection box for detecting sperm chromatin in the prior art has the following disadvantages: 1) the composition is complex, consisting of 3-4 components, wherein each component is prepared from a variety of reagents, and the experimental steps in the detection process are cumbersome and costly. 2) can not be detected by flow cytometer
Most of the existing products can only be stained and observed through a fluorescent microscope, only the number of sperm nucleus fragments can be observed, and the degree of fragmentation and the proportion of immature sperm cannot be analyzed; 3) Even if the existing products can be detected by flow cytometry However, there are many components in the product, the sample processing is complicated, and the subsequent data analysis can only be carried out by its unique software, which cannot be analyzed by conventional flow software

Method used

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Examples

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Comparison scheme
Effect test

Embodiment 1

[0049] An embodiment of the present invention proposes a sperm chromatin structure detection kit, which includes: A reagent and B reagent;

[0050] Wherein, the preparation method of A reagent is: hydrochloric acid 1g, Tween-20 0.5g, add PBS 90ml, adjust pH to be 1, constant volume to 100ml, put into sealed reagent bottle, obtain A reagent, and carry out packing.

[0051] The preparation method of the B reagent is as follows: take 10 mg of acridine orange and dissolve it in 100 ml of purified water or distilled water, fully dissolve to obtain the B reagent, and carry out sub-packaging.

[0052] Another embodiment of the present invention proposes the application of a sperm chromatin structure detection kit. The sperm chromatin structure detection kit in Example 1 is used to detect the fragmentation degree of chromatin DNA and the proportion of immature sperm, which includes :

[0053] Take the flow tube, add diluted with PBS to a concentration of 1 × 10 6 100uL of sperm samp...

Embodiment 2

[0057] An embodiment of the present invention proposes a sperm chromatin structure detection kit, which includes: A reagent and B reagent;

[0058] Among them, the preparation method of A reagent is: hydrochloric acid 1g, Tween-20 0.1g, add PBS 90ml, adjust pH to 1, set the volume to 100ml, put into a sealed reagent bottle, obtain A reagent, and carry out packing.

[0059] The preparation method of the B reagent is as follows: 1 mg of acridine orange is weighed and dissolved in 100 ml of purified water or distilled water, fully dissolved to obtain the B reagent, and subpackaged.

[0060] Another embodiment of the present invention proposes the application of a sperm chromatin structure detection kit. The sperm chromatin structure detection kit in Example 2 is used to detect the fragmentation degree of chromatin DNA and the proportion of immature sperm, which includes :

[0061] Take the flow tube, add diluted with PBS to a concentration of 1 × 10 6 100uL of sperm sample per ...

Embodiment 3

[0065] An embodiment of the present invention proposes a sperm chromatin structure detection kit, which includes: A reagent and B reagent;

[0066] Wherein, the preparation method of A reagent is: hydrochloric acid 1g, Tween-20 0.1g, add PBS 90ml, adjust pH to 4, set to 100ml, put into sealed reagent bottle, obtain A reagent, and carry out packing.

[0067] The preparation method of the B reagent is as follows: Weigh 0.1 mg of acridine orange and dissolve it in 100 ml of purified water or distilled water, fully dissolve to obtain the B reagent, and carry out sub-packaging.

[0068] Another embodiment of the present invention proposes the application of a sperm chromatin structure detection kit. The sperm chromatin structure detection kit in Example 3 is used to detect the fragmentation degree of chromatin DNA and the proportion of immature sperm, which includes :

[0069] Take the flow tube, add diluted with PBS to a concentration of 1 × 10 6 200uL of sperm samples per ml; t...

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Abstract

A sperm chromatin structure detection kit and application thereof are provided. The kit includes an A reagent and a B reagent, with the A reagent including HCl, Tween-20 and a phosphate buffer solution, and the B reagent including acridine orange and water. The kit is used in human sperm chromatin DNA staining to determine the chromatin DNA fragmentation level and to calculate the ratio of spermswith DNA fragmentation, and can analyze the ratio of immature sperms through the degree of combination between chromatin and protein.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a sperm chromatin structure detection kit and its application. Background technique [0002] The latest World Health Organization statistics show that among married couples, the average incidence of unwanted infertility is 7%-46%, and it is increasing year by year. Half of the causes of infertility are caused by male factors. Clinically, male infertility is divided into two categories: sexual dysfunction and normal sexual function. The latter can be further divided into azoospermia, oligospermia, asthenia Spermia, asthenospermia, and normal sperm count infertility. The degree of sperm DNA fragmentation is considered to be a new indicator for evaluating semen quality and predicting fertility. Male infertility patients have significantly increased sperm DNA fragmentation, and the integrity of sperm DNA is an effective basis for judging sperm quality. The degree of sperm DNA frag...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N15/14G01N1/38
CPCG01N1/38G01N15/14G01N21/6402
Inventor 倪万茂迟妍妍陈乐芝倪万根倪声雷张鸿张鸿君林鹏程
Owner ZHEJIANG BOZHEN BIOTECH CO LTD
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