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Method for detecting sperm DNA fragments and device thereof

A technology for sperm and debris, which is applied in biochemical cleaning devices, biochemical equipment and methods, enzymology/microbiology devices, etc., and can solve the problems of complex operation, difficult clinical application, and expensive detection instruments.

Inactive Publication Date: 2009-09-09
SHENZHEN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of these methods have the following unilateral or multiple problems: ① expensive detection instruments are required; ② the operation is relatively complicated; ③ the detection scheme (method and reagent) is not perfect; ④ the detection cost is too high to be popularized and applied clinically

Method used

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Examples

Experimental program
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Embodiment Construction

[0044] The present invention will be described in further detail below through specific examples.

[0045] The sperm DNA fragment detection device (i.e. kit) of the present invention comprises a slide coated with a high melting point gel, a sealed tube encapsulated with an aqueous solution of a low melting point gel, a denaturing solution, a lysing solution, a culture solution, a fluorescent staining solution and Mounting solution. The preparation of each reagent is as follows:

[0046] A1. Preparation of coated slides

[0047] 1) Weigh 2.4g of Tris (trishydroxymethylaminomethane) into a beaker, add distilled water to 900ml, stir to dissolve; use 1mol / L HCl to adjust the pH to 7.4, add distilled water to 1000ml to make a Tris buffer.

[0048] 2) Weigh 1 g of high melting point gel agarose, put it in a beaker, add 100 ml of the above-mentioned Tris buffer, and mix.

[0049] 3) Put the suspension obtained in step 2) into a water bath at 100°C to completely melt.

[0050] 4) ...

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Abstract

The invention relates to a method for detecting sperm DNA fragments, comprising the following steps: 1) adjusting sperm concentration; 2) completely melting low melting point gel, and balancing for standby use; 3) mixing the sperm and the gel following proper proportion; 4) absorbing the obtained suspension to solidoid enveloped with high-melting point gel, and adding a cover plate to the suspension to solidify the suspension until the temperature thereof is 2-8 DEG C after standing; 5) removing the cover plate and soaking the enveloped solidoid horizontally into denatured liquid for reaction under the condition of the temperature being lower than the temperature of the low melting point gel; 6) transferring the enveloped solidoid to solution for reaction under the condition of the temperature being lower than the temperature of the low melting point gel; 7) washing the enveloped solidoid; 8) carrying out continuous dehydration following ethanol concentration sequence from low to high, and drying in the air; 9) dyeing; and 10) counting incidence rate of the DNA fragments under a microscope. The device for detecting sperm DNA fragments comprises an enveloped solidoid, low melting point gel aqueous solution, denatured liquid and washing liquid. The outstanding technological effects are: the result is easy to be judged, the operation is easy and clinical promotion and application are easy.

Description

technical field [0001] The invention relates to a method and a device for in vitro detection, in particular to a method and a device suitable for detecting sperm DNA fragments. Background technique [0002] At present, semen analysis is an indispensable part of infertility examination, mainly including sperm density, sperm motility and sperm morphology examination. Examinations were normal, therefore, further research into new infertility assessment indicators is needed. With the development of molecular biology technology, sperm DNA detection methods are gradually mature and perfect, especially more and more researches on sperm DNA fragmentation, the degree of sperm DNA fragmentation can be used as the etiological diagnosis of infertility, recurrent miscarriage and abnormal embryo A reliable indicator of sperm quality. The mechanism of sperm DNA fragmentation may be related to abnormal chromatin packaging, oxygen stress or abnormal apoptosis during spermatogenesis. The i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12M1/34G01N21/64
Inventor 刘瑜
Owner SHENZHEN PEOPLES HOSPITAL
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