Application of HFM1 gene in preparation of diagnostic kit for detecting non-obstructive azoospermia

A detection kit, azoospermia technology, applied in biochemical equipment and methods, microbial determination/examination, DNA/RNA fragments, etc., to achieve the effect of reducing the economic burden of patients and promoting the development policy

Active Publication Date: 2022-05-06
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0007] However, there is no report about the HFM1 mutation site and its use in the genetic diagnosis of non-obstructive azoospermia.

Method used

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  • Application of HFM1 gene in preparation of diagnostic kit for detecting non-obstructive azoospermia
  • Application of HFM1 gene in preparation of diagnostic kit for detecting non-obstructive azoospermia
  • Application of HFM1 gene in preparation of diagnostic kit for detecting non-obstructive azoospermia

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Embodiment 1

[0021] experimental method

[0022] 1. DNA extraction from peripheral blood (TIANamp Genomic DNA Kit, DP: 304)

[0023] 1) In accordance with the relevant national policies and regulations, and on the basis of the consent of the sampling subject, draw 2-5ml of peripheral venous blood from the first family member, put it into an EDTA anticoagulant tube, and store it at -80°C for later use;

[0024] 2) Take 200ul frozen blood, add 20ul Proteinase K solution, mix well

[0025] 3) Add 200ul buffer solution GB, mix thoroughly by inversion, place at 70°C for 10min, the solution should become clear, and briefly centrifuge to remove water drops on the inner wall of the tube cap.

[0026] 4) 4. Add 200 μl of absolute ethanol, shake and mix well for 15 sec. At this time, flocculent sediment may appear, and briefly centrifuge to remove water droplets on the inner wall of the tube cap.

[0027] 5) Add the solution and flocculent precipitate obtained in the previous step into an adsorpti...

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Abstract

The invention relates to a non-obstructive azoospermia detection kit, the kit comprises a primer pair for detecting a genetic marker related to non-obstructive azoospermia, the nucleotide sequence of the genetic marker is the sequence of an HFM1 gene, and the mutation site of the sequence is c.1006 + 1Ggt; t. The invention also provides application of the detection kit in diagnosis of non-obstructive azoospermia and application of the HFM1 gene in preparation of the diagnostic kit for detecting non-obstructive azoospermia. The invention provides a new application of the HFM1 gene, so that an effective way for performing non-obstructive azoospermia gene diagnosis, prenatal gene screening and genetic counseling is provided; the application effect shows that the SNP site of the gene and the detection primer provided by the invention can be effectively used for rapid detection of the HFM1 gene mutation site in clinical patients and fetal villus or amniotic fluid.

Description

technical field [0001] The invention belongs to the field of medical gene diagnosis, and relates to a genetic disease non-obstructive azoospermia pathogenic gene HFM1 mutation site and its application in gene diagnosis. Background technique [0002] Non-obstructive azoospermia (NOA) is generally considered a non-medically controllable cause of male infertility. These patients represent 10% of all infertile men and the cause of their azoospermia is abnormal spermatogenesis. The establishment of in vitro fertilization, with intracytoplasmic sperm injection (ICSI) as the standard of care, has enabled many of these men to successfully father a child by surgically removing sperm from their testicles. [0003] The dideoxy chain termination method is Sanger sequencing: the reaction system includes target DNA fragments, deoxynucleotide triphosphate (dNTP), dideoxynucleotide triphosphate (ddNTP), sequencing primers and DNA polymerase, etc. The heart of the sequencing reaction is th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 张雨翔姚晨成田汝辉白昊威李铮
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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