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Application of SPACA7 gene

A kind of use and genetic technology, applied in the field of non-obstructive azoospermia detection kits, can solve the problems of cumbersome PCR operation and many primers, and achieve the effect of simple operation, short time-consuming, and beneficial to clinical testing

Inactive Publication Date: 2018-11-23
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Shenzhen Second People's Hospital has applied for a publication number of CN105176976A, titled "A primer pair for detecting genetic markers associated with non-obstructive azoospermia" and a publication number of CN106170562A, titled "A detection and identification A kit for genetic markers related to azoospermia” and patent applications for inventions such as “a genetic marker related to non-obstructive azoospermia” with the publication number CN104531683A, which are characterized by the use of high-throughput sequencing methods to screen Different primer pairs are used to detect the mutation sites of genes in patients with azoospermia, but the disadvantage is that more primer pairs are required, and the PCR operation is cumbersome

Method used

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  • Application of SPACA7 gene
  • Application of SPACA7 gene
  • Application of SPACA7 gene

Examples

Experimental program
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Effect test

experiment example 1

[0035] Experimental example 1: RT-PCR detection of mRNA expression of SPACA7 gene disappears in non-obstructive azoospermia.

[0036] (1) Sample information:

[0037] The information of 44 patients with non-obstructive azoospermia and 30 cases of normal fertility is shown in Table 1:

[0038] Table 1. Sample information (1-44 are patients with non-obstructive azoospermia, 45-74 are normal fertile people)

[0039]

[0040]

[0041]

[0042] (2) Method:

[0043] 1. Sample RNA extraction:

[0044] The total RNA of 74 cases of semen samples was extracted according to the instructions of the Trizol reagent (Invitrogen Company, the U.S.), and the steps were as follows:

[0045] ①Place 1ml of semen sample in a sterile centrifuge tube and centrifuge at 8000g for 5 minutes;

[0046] ② Discard the supernatant and seminal plasma, add TRIzol to the precipitate, mix well, and place at room temperature for 5 minutes to fully lyse the sample;

[0047]③Centrifuge at 12000g at 4°...

experiment example 2

[0075] Experimental Example 2: Immunohistochemical detection of the protein encoded by the SPACA7 gene lost expression in non-obstructive azoospermia.

[0076] 1. Objective: To detect the expression of protein encoded by SPACA7 gene in non-obstructive azoospermia.

[0077] 2. Methods: Formalin-fixed paraffin-embedded tissue sections were dewaxed with xylene, and 3% hydrogen peroxide was used to eliminate endogenous peroxidase activity and 10% bovine serum albumin to prevent non-specific binding. The specimen was added with a primary antibody (Abcam, MA, USA), incubated overnight at 4°C, and the antibody diluent was used instead of the primary antibody to obtain a negative control. Thereafter, sections were incubated at 37°C in MaxVision TM HRP-Polymer anti-Mouse IHC Kit (Maixin, Fujian, China) was treated for 15-20 minutes, soaked in 3-amino-9-ethylcarbazole, counterstained with Mayer's hematoxylin, dehydrated, and finally precipitated as crystals. The result is as figure...

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Abstract

The invention discloses application of an SPACA7 gene. The SPACA7 gene is used for preparing a product for detecting non-obstructive azoospermia. The product comprises a pair of primers for specifically amplifying the SPACA7 gene; the sequence of an upstream primer is shown as SEQ ID NO: 1 and the sequence of a downstream primer is shown as SEQ ID NO: 2. The SPACA7 gene provides a new detection index for detecting the non-obstructive azoospermia. The distinctive detection between normal bearers and azoospermia patients can be achieved by using only a pair of primers through the RT-PCR detection; the whole operation process is simpler, consumes shorter time and is beneficial to clinical testing.

Description

technical field [0001] The invention belongs to the field of genetic engineering technology and reproductive medicine, in particular to a non-obstructed azoospermia detection kit. Background technique [0002] According to the statistics of the World Health Organization, infertility accounts for about 20% of couples of childbearing age, of which male factor causes infertility accounts for about 50%. Male infertility has brought a heavy burden to individuals, families, and society. The World Health Organization announced that infertility, cardiovascular disease, and tumors are listed as one of the three major diseases that affect human life and health today. With the improvement of social industrialization and the acceleration of the pace of life, coupled with factors such as environmental pollution and bad living habits, the quality of male semen is declining, and the number of male infertility patients such as azoospermia, oligospermia and asthenospermia is increasing year ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/156
Inventor 张菁华刘井清张丹唐爱发
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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