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Sperm DNA fragmentation detection kit

A technology for detecting kits and sperm, which is applied in the preparation of test samples, measuring devices, and the determination/inspection of microorganisms, etc. It can solve the problems of unclear tails of sperm, dark background color, and difficult interpretation, etc., so as to facilitate clinical routine application and promotion, simple operation, and the effect of saving manpower and material resources

Active Publication Date: 2014-10-01
BRED LIFE SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, some merchants have launched commercial sperm DNA fragment detection kits on the market, but most of them have some problems, such as: the background color is too dark, the tail of sperm is not clear, and the halo is too small, which makes interpretation difficult, etc.

Method used

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  • Sperm DNA fragmentation detection kit
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Examples

Experimental program
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preparation example Construction

[0087] The preparation method of described coated glass slide is:

[0088] 1) Weigh 1-50 g of low-melting point agarose and place it in a container that can hold 1000 ml, add 50-200 ml of purified water, stir in a water bath at 50±5°C to completely dissolve the agarose.

[0089] 2) Add purified water to make up to 1000ml, stir and mix well, and keep warm in a water bath at 50±5°C.

[0090] 3) Place the solution in step 2) on the surface of the glass slide while it is still hot, and dry it.

[0091] The preparation method of the fusible gel is:

[0092] 1) Weigh 1-50g of agarose, put it in a beaker, put it in a container that can hold 1000ml, add 50-200ml of purified water, and stir in a water bath at 50±5°C to completely dissolve the agarose.

[0093] 2) Add purified water to make up to 1000ml, stir and mix, and keep warm in a water bath at 50±5°C for 2 hours.

[0094] 3) While hot, divide the solution in step 2) into 0.5ml sample tubes, 0.14ml per tube.

[0095] The prepa...

Embodiment 1

[0117] This embodiment provides a sperm DNA fragment detection kit, comprising:

[0118] coated slides;

[0119] fusible gel;

[0120] A liquid;

[0121] liquid B;

[0122] Wright's stain;

[0123] Wright's buffer;

[0124] SCD preservation solution.

[0125] The coated glass slide is a glass slide coated with an aqueous solution of 0.5% low melting point agarose; the fusible gel is an aqueous solution containing 0.25% agarose; the A solution is a solution containing 0.05% sodium chloride , 0.55% aminoacetic acid, 0.25% glacial acetic acid and 0.045% hydrogen peroxide aqueous solution; the B liquid is containing 0.25% SDS, 2.422% Tris, 0.125% disodium edetate dihydrate, 0.15% Tween 20 And the aqueous solution with a pH value of 7.2~7.4; the Wright’s staining solution is a methanol solution containing 0.1% Wright’s pigment and 0.03% Gibson’s pigment; the Wright’s buffer is 0.03mol / L with a pH value of 6.2~6.4 Phosphate buffer saline; the SCD preservation solution is an a...

Embodiment 2

[0155] This embodiment provides a sperm DNA fragment detection kit, comprising:

[0156] coated slides;

[0157] fusible gel;

[0158] A liquid;

[0159] liquid B;

[0160] Wright's stain;

[0161] Wright's buffer;

[0162] SCD preservation solution.

[0163] The coated glass slide is a glass slide coated with an aqueous solution of 2% low-melting point agarose; the fusible gel is an aqueous solution containing 1% agarose; the A solution is a solution containing 0.2% sodium chloride , 2.2% aminoacetic acid, 0.10% glacial acetic acid and 0.18% hydrogen peroxide aqueous solution; the B liquid is containing 1% sodium dodecyl sulfate (SDS), 10% Tris, 0.5% dihydrate ethylenediaminetetraacetic acid Disodium, 0.6% Tween 20 and the aqueous solution of pH value 7.6~8.0; Described Wright's staining solution is the methanol solution containing 0.4% Wright's pigment, 0.12% Gibson's pigment; Described Wright's buffer solution is the pH value 0.12mol / L phosphate buffer solution of 6....

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Abstract

The invention provides a sperm DNA fragmentation detection kit. The kit comprises an enveloped glass slide, a fusible gel, an A solution, a B solution, a Wright stain, a Wright buffer solution and a SCD (sperm chromatin diffusion) storage solution. The invention further provides a method for detecting sperm DNA fragmentation by using above kit. The reagent structure is optimized according to the detection method so that the reagent is stable and reliable; the operation of detecting by using the reagent is simple and free from special detection instruments, the clinical conventional application and popularization are facilitated; by adopting the unique SCD storage solution, the sperm sample can be effectively stored, the DNA fragmentation rate cannot be changed after being cryopreserved at -20 DEG.C within two weeks, the required sample can be easily stored through the SCD storage solution, the detection can be performed without the limitation of time and number, and large number of manpower and material resources are saved.

Description

technical field [0001] The invention belongs to the category of in vitro detection reagents, in particular to a detection kit for sperm DNA fragments. Background technique [0002] The degree of fragmentation of sperm DNA reflects the integrity of sperm genetic material. Sperm chromatin diffusion assay is one of the main methods to detect sperm DNA fragmentation. Sperm with complete DNA diffuses to form a characteristic halo after undergoing denaturation and removal of nucleoprotein, while sperm with DNA fragments do not produce this characteristic halo, and the DNA of the sperm is judged according to the presence or absence and size of the halo Completeness. [0003] At present, some merchants have launched commercial sperm DNA fragment detection kits on the market, but most of them have some problems, such as: the background color is too dark, the sperm tail is not clear, and the halo is too small, which makes interpretation difficult. Contents of the invention [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCG01N1/2813G01N1/30G01N33/487
Inventor 程浩陶思恩邓少君张翔庄学敏钟彩颜
Owner BRED LIFE SCI TECH
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