Application of PKP3 gene methylation in sperm activity detection agent and kit

A detection kit, sperm motility technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of accurate evaluation of sperm motility, less methylated regions, etc.

Pending Publication Date: 2020-02-28
深圳市龙华区人民医院
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Problems solved by technology

[0004] However, there are very few methylated regions related to sperm motility reported so far, and it is difficult to accurately evaluate sperm motility. Therefore, it is necessary to identify more methylated regions related to sperm motility, so as to provide a basis for the application of subsequent diagnosis and treatment. more basis

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  • Application of PKP3 gene methylation in sperm activity detection agent and kit
  • Application of PKP3 gene methylation in sperm activity detection agent and kit
  • Application of PKP3 gene methylation in sperm activity detection agent and kit

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Embodiment Construction

[0015] The principles and features of the present invention are described below in conjunction with the accompanying drawings, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.

[0016] 1. Sperm separation

[0017] Seven asthenospermia volunteers (group A) and eight healthy volunteers (group N) were recruited, and sperm samples were obtained from the volunteers for genome-wide sequencing of methylation levels. Computer-assisted sperm analysis (CASA) was performed using a Sperm Class Analyzer, and sperm morphology was assessed by Diff-Quik rapid staining. Sperm cells with different vigor were separated by the following method: centrifuged at 300×g for 20 min to prepare discontinuous density gradient fractions (divided into 47.5, 57, 76 and 95% concentration layers), and from the 95% concentration layer (fraction a) High motility sperm cells were collected, and low motility sperm cells were c...

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Abstract

The invention relates to application of methylation level of a human PKP3 gene promoter region in preparing a detection agent for preparing sperm activity. The human PKP3 gene promoter region is a region corresponding to space from 392017th to 394717th of a human 11th chromosome. Analysis, research and verification on the basis of whole genome methylation sequencing results finds that the methylation level of the human PKP3 gene promoter region can serve as a molecular marker to detect the sperm activity, and when all methylation sites of the region are not completely methylated, it can be thought that activity of corresponding sperms is weak. The molecular marker can provide assistant evaluation information for diagnosis of asthenozoospermia and selection of sperms before artificial fertilization.

Description

technical field [0001] The invention relates to the field of human reproductive health, more particularly, relates to the application of the methylation level of the promoter region of the human PKP3 gene in the preparation of a detection agent for detecting sperm motility, and a detection kit for detecting sperm motility. Background technique [0002] More than 40% of infertile men were detected to suffer from asthenospermia. The etiology of asthenospermia is multifactorial, for example, it may be closely related to endocrine disorders, environmental factors, life experiences and even aging. Gene knockout and mutation studies in mice have shown that mutations in some genes are associated with altered sperm motility, but none of these genes have been shown to be associated with sperm motility in human studies. Although nearly 50% of infertility cases are believed to be caused by genetic defects, in some cases of male infertility, genetic factors do not appear to be the main...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/154
Inventor 杜野桂耀庭邓琼马倩杨波
Owner 深圳市龙华区人民医院
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