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Non-obstructive azoospermia auxiliary diagnosis gene detection kit

A technology for azoospermia and auxiliary diagnosis, which can be used in the determination/examination of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc. It can solve the problems of difficult clinical diagnosis of non-obstructive azoospermia and achieve high specificity.

Inactive Publication Date: 2019-06-28
YINFENG JILIN BIOLOGICAL ENG TECH CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In order to solve the problem of difficult clinical diagnosis of non-obstructive azoospermia, the present invention provides a gene detection kit for auxiliary diagnosis of non-obstructive azoospermia

Method used

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  • Non-obstructive azoospermia auxiliary diagnosis gene detection kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] The gene detection kit for auxiliary diagnosis of non-obstructive azoospermia provided by the present invention includes specific PCR primers, PCR buffer, dNTPs and DNA polymerase, wherein the specific PCR primers are the 69826819th position in the TEX11 gene for detecting male infertility A set of primers specific to the mutation site.

[0016] Wherein, the PCR buffer adopts 10 mmol / L Tris-Cl buffer, and the pH is 8.3-8.8.

[0017] DNA polymerase (DNA polymerase) is an important enzyme for cell replication DNA. DNA polymerase, using DNA as a replication template, is an enzyme that copies DNA from the 5' end to the 3' end. The main activity of DNA polymerase is to catalyze the synthesis of DNA and its complementary activities.

[0018] The upstream primer sequence of the specific PCR primer is shown in SEQ ID NO:1, and the downstream primer sequence is shown in SEQ ID NO:2. See Table 1 for sequence information.

[0019] Table 1

[0020]

[0021] Target gene name...

Embodiment 2

[0033] Example 2 The method of using the gene detection kit for auxiliary diagnosis of non-obstructive azoospermia

[0034] The gene detection kit for auxiliary diagnosis of non-obstructive azoospermia was used to detect the 69826819th mutation site in the TEX11 gene of male infertility. The specific operation process is as follows:

[0035] 1. PCR amplification

[0036] The PCR reaction conditions were: pre-denaturation at 95°C for 5min; denaturation at 95°C for 30s, annealing at 68°C for 2min30s, 30 cycles; extension at 72°C for 10min.

[0037] 2. Identification of PCR products: PCR products were identified by agarose electrophoresis analysis, and sent to Changchun Kumei Technology Co., Ltd. for sequencing after PCR identification.

[0038] 3. Observe the condition of the electrophoresis strip under ultraviolet light, and judge whether the sample contains a pathogenic mutation gene according to the condition of the electrophoresis strip.

[0039] Take the patient's blood a...

Embodiment 3

[0040] Embodiment 3 clinical trials

[0041] Screening of 30 patients with clinically confirmed non-obstructive azoospermia (the cause of these 30 patients is all caused by the mutation of the 69826819th gene in the TEX11 gene of male infertility, the position of the mutated gene is at 69826819, showing T >C homozygous mutation, the SNP of the two gene loci after the mutation is GG), using the non-obstructive azoospermia auxiliary diagnosis gene detection kit of embodiment 1, referring to the use method of embodiment 2 for these 30 patients Detection of genetic mutation sites. Observation of electrophoresis bands under ultraviolet light showed that there were no bands of 100 items, which indicated that the sample DNA template contained a mutant gene at the 69826819 mutation site, and the detection rate of gene mutation was 100%.

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Abstract

The invention discloses a non-obstructive azoospermia auxiliary diagnosis gene detection kit, relates to the field of azoospermia diagnosis, and solves the problem of difficult clinical diagnosis of male infertility. The non-obstructive azoospermia auxiliary diagnosis gene detection kit comprises a specific PCR primer, a PCR buffer solution, dNTPs and DNA polymerase, wherein the specific PCR primer is a group of specific primers for detecting the 69826819 mutation site in the non-obstructive azoospermia TEX11 gene. A specific primer SEQ ID NO: 1 and a specific primer SEQ ID NO: 2 are designed,the specific primers have higher specificity, the non-obstructive azoospermia auxiliary diagnosis gene detection kit consisting of the specific primers is adopted to detect the 69826819 mutation sitein the TEX11 gene of a male patient, and the detection rate of the gene mutation is 100%.

Description

technical field [0001] The invention relates to the technical field of diagnosis of azoospermia, in particular to a gene detection kit for auxiliary diagnosis of non-obstructive azoospermia. Background technique [0002] Male infertility refers to a couple who have a normal sex life after marriage and do not use any contraceptive measures, and the wife is infertile due to the man's reasons. Male infertility seriously affects men's health. According to the World Health Organization survey, about 8% of couples in the childbearing age suffer from infertility. Based on this, it is estimated that about 50 million to 80 million people in the world suffer from infertility problems , and showed an increasing trend year by year. Among them, the proportion of infertility caused by male reasons is generally not less than 50%. Male infertility can be divided into absolute infertility and relative infertility according to clinical manifestations; it can be divided into primary infertil...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
Inventor 刘睿智张红国魏本杰王瑞雪李博苏鹤
Owner YINFENG JILIN BIOLOGICAL ENG TECH CO LTD
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