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Kit for detecting idiopathic azoospermia and gene panel

A kit and azoospermia technology, which are applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems that have not been reported, reduce workload, increase detection rate, and save work the effect of time

Pending Publication Date: 2019-05-28
XUKANG MEDICAL SCI & TECH (SUZHOU) CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, there are no reports about the test kit and gene panel for detecting idiopathic azoospermia of the present invention

Method used

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  • Kit for detecting idiopathic azoospermia and gene panel
  • Kit for detecting idiopathic azoospermia and gene panel
  • Kit for detecting idiopathic azoospermia and gene panel

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Experimental program
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Effect test

Embodiment

[0026] The present invention uses the following gene Panel to perform exons on blood samples from 100 idiopathic azoospermia patients and 300 Chinese Han azoospermia patients and blood samples provided by qualified volunteers who donated sperm from the Shanghai Human Sperm Bank Sequencing, the specific steps are as follows:

[0027]

[0028]

[0029] 1DNA extraction and detection

[0030] Genomic DNA of all samples was extracted from EDTA-anticoagulated peripheral whole blood using QIAamp DNA Blood Midi Kit (Qiagen). The DNA was quantified using OD measurement (NanoDrop, Thermo Scientific) and used for library preparation (approximately 3 μg) according to Agilent's requirements.

[0031] 2 Tn5 transposase-based linker insertion and fragmentation

[0032] (1) Configure the reaction system on ice:

[0033]

[0034]

[0035] (2) Blow and suck and mix well, then spin and centrifuge;

[0036] (3) Incubate at 50~60℃ in PCR instrument for 5 minutes;

[0037] (4) Put the reaction system on ice ...

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Abstract

The invention relates to the field of gene detection technology, in particular to a kit for detecting idiopathic azoospermia and a gene panel. A small amount of (3 ml) peripheral blood from a patientis utilized to extract DNA, gene mutation of the patient is detected through library-establishing, high-throughput sequencing, etc., and through bioinformatics analysis, such as mutation frequency, mutation property, biological harm and conservative property, evaluation is performed to find 30 new morbigenous genetic mutations in the patient. Only 3 ml of peripheral blood of the patient is neededfor detection and 30 detection sites are covered, thereby improving the detection rate of the mutation, and 30 sites are carried out in one reaction system, to greatly reduce the workload, improve work efficiency, and save work time.

Description

Technical field [0001] The invention relates to the technical field of gene detection, in particular to a kit and gene panel for detecting idiopathic azoospermia. Background technique [0002] Azoospermia is the most serious condition that causes male infertility. The core feature is the absence of sperm in the ejaculated semen. According to statistics, it affects about 1% of men in the world. Azoospermia has a high degree of heterogeneity in etiology. Many studies have confirmed that genetic abnormalities are an important cause of azoospermia, accounting for about 15% of all azoospermia cases. Known causes include Yq microdeletion and Klinefelter synthesis. Levy and so on. In 2015, Yatsenko, A.N. and their colleagues found that a rare mutation of TEX11 caused spermatogenesis disorders in a group of sporadic azoospermia patients, and published their results in the top academic journal of medical research "The Lancet". During the same period, there were many studies. For example...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
Inventor 孙斐陈石涛郑小国王贵栓
Owner XUKANG MEDICAL SCI & TECH (SUZHOU) CO LTD
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