Kit for diagnosing asthenospermia through piR-57660 and application

A technology of pir-57660 and diagnostic kits, applied in the field of medical diagnosis, can solve problems such as difficult storage of samples and complicated steps

Pending Publication Date: 2021-07-16
NANJING YOUZHIYUAN PHARMACEUTICAL TECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the diagnosis mainly relies on routine semen analysis, which is a clinical indicator for judging male fertility and diagnosing asthenospermia, but has the disadvantages of complicated steps and difficult storage of samples

Method used

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  • Kit for diagnosing asthenospermia through piR-57660 and application
  • Kit for diagnosing asthenospermia through piR-57660 and application

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Embodiment Construction

[0032] The present invention is illustrated below with reference to specific examples. Those skilled in the art can understand that these examples are only used to illustrate the present invention and do not limit the scope of the present invention in any way.

[0033] Quantitative detection experiment of piRNA in seminal plasma, the specific steps are:

[0034] (1) Collection of seminal plasma from normal persons and certain patients

[0035] (2) First extract 100 μl of seminal plasma total RNA using Trizol reagent.

[0036] (3) RNA reverse transcription reaction to obtain cDNA. The reverse transcription reaction system includes: 1 μl AMV, 2 μl dNTP, 4 μl AMV buffer, 5 μl RRI, 1 μl specific primer, 9.5 μl DEPC, 2 μl RNA. The reaction steps are 15 minutes at 16°C, 60 minutes at 42°C, 5 minutes at 85°C, and storage at 4°C.

[0037] (4) Perform Quantitative real-time PCR reaction. The reaction system is: 0.3 μl rTaq, 0.4 μl DNTP, 1.2 μl Mg2+, 2 μl 10× buffer, 14.5 μl ddH2O,...

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Abstract

The invention provides an asthenospermia marker as well as a detection method and a kit thereof. The asthenospermia marker provided by the invention comprises ribonucleic acid piR-57660 which stably exists in seminal plasma of a subject, can be detected and interacts with Piwi protein. The invention also provides a kit comprising a tool or element for detecting the asthenospermia marker. The combination, the method and the kit provided by the invention can be used for assisting in diagnosis and differential diagnosis of asthenospermia, prediction of occurrence and relapse of disease complications, curative effect evaluation, screening of active pharmaceutical ingredients, drug effect evaluation and the like, and have the advantages of high sensitivity, low detection cost, convenience in material taking, easiness in sample storage and the like; the method can be widely applied to general survey of asthenospermia and other related work, and becomes an effective means for early diagnosis of asthenospermia.

Description

technical field [0001] The invention belongs to the field of medical diagnosis and relates to the separation, qualitative and quantitative analysis of piRNA molecules in human seminal plasma. More specifically, the present invention is a method for detecting piR-57660 in the seminal plasma of patients with asthenospermia, and can diagnose asthenospermia in vitro through the change of piR-57660 in the seminal plasma of patients with asthenospermia. Background technique [0002] In recent years, due to the influence of various factors, the number of infertility patients has been increasing. Infertility will seriously affect the patient's body, mind and quality of life, and bring a heavy burden to the family. With the increasing pressure of people's life, environmental pollution, life and diet have undergone major changes, and these factors are closely related to the occurrence of infertility. [0003] Among infertile patients, most of them are oligoasthenospermia patients, wh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 赵艺陈骁锐骆阳夏玉贵屠盛林巧芳高娜
Owner NANJING YOUZHIYUAN PHARMACEUTICAL TECHNOLOGY CO LTD
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