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An antigen for detecting sperm antibody, its expression vector, its kit and detection method

A technology for expressing vectors and antibodies, which is applied in the field of reproductive medicine and immune applications, can solve the problems of high false positive rate, unreliable sensitivity and accuracy, and low positive rate, and achieve the effect of less specimen consumption and reliable test results

Active Publication Date: 2021-06-04
东莞兰卫医学检验实验室有限公司
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

However, the commonly used ELISA kits in China have a high false positive rate or a low positive rate, and the sensitivity and accuracy of the domestic kits are unreliable, which cannot meet the clinical needs for infertility testing, so a new, cheap one is urgently needed. Anti-sperm antibody test kit with reliable test results to meet clinical needs

Method used

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  • An antigen for detecting sperm antibody, its expression vector, its kit and detection method
  • An antigen for detecting sperm antibody, its expression vector, its kit and detection method
  • An antigen for detecting sperm antibody, its expression vector, its kit and detection method

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Embodiment 1

[0057] An antigen used to detect sperm antibodies is a polypeptide encoded by the gene fragment of the PB functional region of the recombinant IZUMO gene, its amino acid sequence has the sequence shown in SEQ ID NO: 1, and its gene sequence has the sequence shown in SEQ ID NO: 2 .

Embodiment 2

[0059] A kind of expression vector used for detecting the antigen of sperm antibody in embodiment 1, the name of this expression vector is pET30a-IZUMO-His, the construction diagram of this expression vector is shown in the appendix figure 1 .

Embodiment 3

[0061] A method for constructing an expression vector for detecting an antigen of a sperm antibody according to embodiment 2, comprising the following steps:

[0062] Step 1, synthesize the gene fragment of the PB functional region of the recombinant IZUMO gene: obtain the PB gene sequence by gene synthesis and insert a His tag at the 3' end to obtain the gene fragment with the sequence shown in SEQ ID NO:1;

[0063] Step 2, amplification of the PB gene fragment: using the gene fragment obtained in Step 1 as a template, perform PCR amplification to obtain the above-mentioned expression vector for the antigen used for detecting sperm antibodies.

[0064] Wherein, in step 2, the conditions for PCR amplification are: pre-denaturation at 94°C for 5 minutes, denaturation at 94°C for 40 seconds, annealing at 60°C for 30 seconds, extension at 72°C for 45 seconds, 30 cycles, and extension at 72°C for 10 minutes. Wherein, the reaction system is shown in Table 1.

[0065] Table 1 React...

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Abstract

The invention relates to the technical fields of reproductive medicine and immune application, in particular to an antigen for detecting sperm antibodies, an expression carrier thereof, a kit and a detection method thereof. An antigen used for detecting sperm antibodies of the present invention is a polypeptide encoded by a gene fragment of the PB functional region of the recombinant IZUMO gene, and its sequence has the sequence shown in SEQ ID NO:1. The antigen used to detect sperm antibody is the key protein of sperm acrosome reaction, which determines the success of fertilization, and the antibody against this antigen is one of the important causes of immune infertility. The ELISA kit for sperm antibody detection can detect anti-sperm antibodies, overcomes the existing defects of low sensitivity and low specificity, and is convenient, sensitive, requires less specimen, can be tested in large quantities, and can be popularized and scaled in ordinary laboratories Etc. Therefore, the present invention is expected to be used in the screening and diagnosis of antisperm antibodies in infertile patients.

Description

technical field [0001] The invention relates to the technical fields of reproductive medicine and immune application, in particular to an antigen for detecting sperm antibodies, an expression carrier thereof, a kit and a detection method thereof. Background technique [0002] Infertility refers to a state in which a couple lives together for at least one year after marriage, has a normal sex life, and has not taken any contraceptive measures and is infertile. According to the survey data conducted by the World Health Organization (WHO) in 33 centers in 25 countries using standardized infertility diagnostic methods in the late 1980s, there are about 60 million to 80 million couples in the world who are infertile. Due to factors such as environment, hormones and related diseases, the number of infertile couples is increasing day by day, and about 8% to 12% of couples in the world have experienced infertility. Relevant studies have shown that the fertility of couples of childb...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C12N15/70C12N15/66G01N33/68G01N33/577
CPCC07K14/47C12N15/66C12N15/70G01N33/577G01N33/68G01N2333/46
Inventor 徐军发王鑫徐健庭
Owner 东莞兰卫医学检验实验室有限公司
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