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Secreting type IgA endomethal antibody detection kit and use method of secreting type IgA endomethal antibody detection kit

An endometrial and antibody detection technology, applied in the field of enzyme immunoassay, can solve problems such as damage to the endometrium, dysfunction of the physiological barrier network in the internal environment, and failure of gestational egg implantation.

Inactive Publication Date: 2013-07-10
TIANJIN BAODI HOSPITAL
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  • Abstract
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AI Technical Summary

Problems solved by technology

The normal position of the endometrium has no antigenicity to the body, but the female reproductive tract infection changes the internal environment, destroys the physiological barrier, and disrupts the network function of idiotype antibodies and anti-idiotype antibodies in women. At the same time, it can also make the immune system Damage and damage to the endometrium will cause the body to use its own endometrial tissue as an antigen to stimulate the body to produce anti-endometrial antibodies, and the ectopic endometrium can also be used as an antigen to stimulate the body’s immune system to produce specific anti-endometrial antibodies Antibodies, which can bind to antigens in the endometrium, undergo antigen-antibody reactions, activate the complement system, and undergo a series of immune reactions, resulting in insufficient glycogen secretion of nutritional embryos. At the same time, anti-endometrial antibodies are often associated with endometriosis Symptoms coexist, can inhibit ovulation, interfere with the transportation of sperm and eggs, hinder the combination of sperm and eggs, affect the development of early embryos, interfere with embryo implantation, etc., eventually lead to the failure of implantation of pregnant eggs and cause infertility in patients

Method used

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  • Secreting type IgA endomethal antibody detection kit and use method of secreting type IgA endomethal antibody detection kit
  • Secreting type IgA endomethal antibody detection kit and use method of secreting type IgA endomethal antibody detection kit
  • Secreting type IgA endomethal antibody detection kit and use method of secreting type IgA endomethal antibody detection kit

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Embodiment Construction

[0009] 1. Preparation of anti-endometrium antibody ELISA plate

[0010] Dilute the endometrial antigen with coating solution to 1:200 times, add 100 μl to the microwells of the coated microwell reaction plate, add 350 μl / well of phosphate buffer containing 1% bovine serum albumin, and block at room temperature for 2 hours , overnight at 4°C. After shaking off the liquid in the hole, wash 3 times with washing solution, 3 minutes each time.

[0011] 2. Preparation of enzyme-labeled antibody

[0012] (1) Dissolve 5mg of horseradish peroxidase in 0.5ml0.2mol / L pH5.6HAC-NaAc buffer solution, add 0.1mol / L NaIO 4 0.25ml, 30 minutes at 4°C.

[0013] (2) Add 0.5ml of 2.5% ethylene glycol, 30min at room temperature.

[0014] (3) Add 5-10 mg of secondary anti-endometrial antibody, adjust the pH of the reaction solution to about 9.0 with 1.0 mmol / L pH9.5 carbonate buffer, and overnight at 4°C.

[0015] (4) Add NaHB 4 Mix 0.1ml, and dialyze against 0.01mol / L pH7.4 PBS at 4°C for 2 ho...

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Abstract

The invention discloses a secreting type IgA endomethal antibody detection kit and a use method of the secreting type IgA endomethal antibody detection kit, belonging to the technical field of enzyme immunoassay. The use method of the kit disclosed by the invention comprises following steps of: breaking a disulfide bond (-S-S-) structure of a secreting type IgA endomethal antibody polymer in cervical wecretion by N-acetyl-L-cysteine, converting into a soluble monomer, adding the soluble monomer onto an endomethal antibody detection elisa plate, detecting the secreting type IgA endomethal antibody by an enzyme-linked immuno sorbent assay (ELISA) method, and carrying out warm bath, washing and IgG-HRP adding, thus reacting and washing the two parts, adding an enzyme substrate for color development. The invention provides the detection method which is simple, quick, sensitive, exact and cheap in pretreatment, is suitable for the detection of the secreting type endomethal antibody in cervical wecretion, and can be taken as an important examine index for infertility.

Description

technical field [0001] The invention relates to a secreted IgA anti-endometrium antibody enzyme immunoassay kit and a detection method thereof, which belong to the technical field of enzyme immunoassay. Background technique [0002] Infertility is divided into infertility and infertility. According to the cause of occurrence, it can be divided into organic, immune, iatrogenic, infectious, and physiological infertility. According to whether there is a history of pregnancy, it can be divided into Primary infertility and secondary infertility. There are also 10% to 20% of patients whose cause is unknown, and in such patients, immune factors account for 30 to 40% and play a role that cannot be ignored. Therefore, the clinical diagnosis and treatment of immune infertility has been paid more and more attention. It will help clinical diagnosis and treatment and bring good news to patients. Anti-endometrial antibodies (AEAb) mainly include three types: IgG, IgM, and IgA. Pregnancy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/531
Inventor 李立和王彦平李铮
Owner TIANJIN BAODI HOSPITAL
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