NANOS2 marker for predicating or diagnosing azoospermia or oligospermia and application thereof

An azoospermia and marker technology, applied in the field of biomedicine, can solve the problems of slow effect, long detection time, difficulty in patient prediction, diagnosis, treatment and prognosis judgment, and achieve high accuracy and specificity, and the method is simple. Effect

Inactive Publication Date: 2018-11-16
YUANJIAN BIOTECH SHANGHAI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current treatment methods for this disease include drug therapy based on drugs for invigorating the kidney, strengthening yang and promoting sperm production, or surgical treatment, but the effect is poor and the effect is slow, so the treatment effect is not ideal
Male spermatogenesis disorder is receiving increasing attention, but due to the complexity of the spermatogenesis proces

Method used

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  • NANOS2 marker for predicating or diagnosing azoospermia or oligospermia and application thereof
  • NANOS2 marker for predicating or diagnosing azoospermia or oligospermia and application thereof
  • NANOS2 marker for predicating or diagnosing azoospermia or oligospermia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Preparation of NANOS2 polyclonal antibody

[0031]Firstly, the NANOS2 gene was amplified by PCR method, and then the target fragment was recovered by agarose gel electrophoresis, followed by enzyme digestion and overnight ligation, and the NANOS2 gene was cloned into the pGEX4T3 plasmid, and the NANOS2 / pGEX4T3 plasmid was transformed into Escherichia coli BL21 cells. BL21 Escherichia coli cells express the purified NANOS2-GST fusion protein, and the classic IPTG-induced protein expression method is used to induce the expression of the target protein. Afterwards, Escherichia coli was lysed and GST-beads were used to purify the NANOS2-GST fusion protein. Rabbits were immunized with the NANOS2-GST fusion protein to prepare specific NANOS2 polyclonal antibodies, and the protein function was tested in vitro. Wherein, the NANOS2 protein used in the preparation of the NANOS2 polyclonal antibody is a full-length protein.

[0032] Wherein, the synthetic method of NA...

Embodiment 2

[0042] Example 2: Low expression of NANOS2 in human SCO azoospermia

[0043] The present invention adopts immunofluorescent staining method to stain the paraffin sections of normal testis and SCO azoospermia testis, and the staining results are as follows: Figure 4 As shown, there are 3 to 6 cells expressing NANOS2 protein around each lumen in the normal testis (left picture), that is, cells with positive green fluorescence (as shown by the arrow in the figure); while SCO has no There are no cells expressing NANOS2 protein in the lumen of the testis with sperm (right picture), that is, the green fluorescence is a negative signal.

[0044] The normal testes and SCO type azoospermic testes were divided into six groups, such as Figure 5 Shown; Among them, the normal testis group consists of two groups, namely control 1 and control 2, and the SCO azoospermia testis group is four groups, respectively SCO1-4. Take 5 seminiferous tubules in each group for statistics, and detect t...

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Abstract

The invention discloses an NANOS2 marker for predicating or diagnosing azoospermia or oligospermia. The NANOS2 marker can be DNA or mRNA of NANOS2 or protein coded by the NANOS2. The invention furtherprovides an NANOS2 polyclonal antibody, and an in vitro diagnostic reagent prepared according to the NANOS2 marker. The in vitro diagnostic reagent is used for predicating or diagnosing azoospermia or oligospermia, has the advantages of high convenience and rapidness, has high accuracy and specificity, can be applied to large-scale screening, can further serve as a clinical detection means, and particularly provides a novel approach for the diagnosis and detection of SCO azoospermia.

Description

technical field [0001] The invention relates to the technical field of biomedicine, and relates to a NANOS2 marker for predicting and diagnosing azoospermia or oligospermia and its application, specifically, the application of the protein encoded by the NANOS2 gene and its antibody in the diagnosis of SCO azoospermia. Background technique [0002] Male azoospermia and oligospermia are one of the main causes of male infertility. The causes of male infertility and oligospermia are complex, some are due to congenital deficiencies, some are caused by acquired disorders, including abnormalities in chromosome number and structure, microbial infections, and drug effects. The current method for the treatment of this disease includes drug therapy based on drugs for invigorating the kidney, strengthening yang and promoting sperm production, or through surgical treatment, but the effects are poor and slow to take effect, so the therapeutic effect is not satisfactory. Male spermatogeni...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577
CPCG01N33/577G01N33/68G01N2800/367
Inventor 杜飞
Owner YUANJIAN BIOTECH SHANGHAI CO LTD
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