Embryo implantation detection kit, and application and use method thereof

A detection kit and embryo implantation technology, applied in the field of kits

Pending Publication Date: 2020-05-08
THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims to solve the problem of how to detect abnormal maternal fetal immune tolerance in the prior art, and proposes an embryo implantation detection kit using decidua NK cells Siglec-7 as a detection index and its application and use method. Expression detection of Siglec-7, the kit can quickly detect abnormal immune tolerance of maternal fetal surface

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  • Embryo implantation detection kit, and application and use method thereof

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Embodiment 1

[0047] The current study found that recurrent unexplained abortion is mainly related to the abnormal immune tolerance of maternal fetal surface. Natural killer cells, or NK cells, account for 80% of uterine lymphocytes and play an important role in uterine pregnancy immunity. Siglec-7, sialic acid-binding immunoglobulin-like lectin-7, is mainly distributed in peripheral blood NK cells and is an important inhibitory receptor on the surface of NK cells.

[0048] We found that Siglec-7 is expressed in human decidua NK cells, and abnormal expression will lead to the destruction of the balance between embryonic trophoblast cells and endometrium. However, there is still a lack of corresponding diagnostic indicators for uterine NK cell function. At the same time, there are limitations in the evaluation of endometrial embryo implantation ability. Therefore, Siglec-7 can be used as a molecular marker to evaluate the implantation ability of endometrial embryos.

[0049]The invention p...

Embodiment 2

[0052] The invention provides an embryo implantation detection kit, which uses the expression degree of decidua NK cells Siglec-7 in a decidua sample as a detection index.

[0053] Further, the embryo implantation detection kit includes reagents for detecting the expression of Siglec-7 in decidua NK cells.

[0054] Specifically, the embryo implantation detection kit includes: PBS buffer, PBST buffer, PBT buffer, 4% paraformaldehyde, tetrahydrofuran, xylene I, xylene II, sodium citrate, citric acid, 30% H 2 o 2 , bovine serum albumin and DAPI.

[0055] The preparation method of the above reagents is: add 3g of sodium citrate, 0.4g of citric acid into 1000ml of distilled water; the routine storage concentration of H2O2 is 30%, and it should be sealed and stored away from light. Currently used; bovine serum albumin is powder, stored at 4°C, self-preparation is required when using: dissolve 0.3g BSA in PBT, and finally dilute to 10ml with PBT, PBT=0.1% Triton X-100 in PBS; store...

Embodiment 3

[0057] The present invention also proposes the detection method of embryo detection kit, specifically comprises the following steps:

[0058] X1. Sample preprocessing

[0059] Take 5 to 15 g of decidua tissue samples, discard the villi on the surface of the decidua, wash with PBS buffer for 2 to 3 times, and fix in 4% paraformaldehyde for 8 to 12 hours.

[0060] X2. Dehydration

[0061] The samples processed in X1 were dehydrated through pure water for 2 hours, 50% ethanol for 2 hours and 75% ethanol in sequence.

[0062] X3. Embedding

[0063] The samples treated in X2 were dehydrated with 75% ethanol and tetrahydrofuran at a volume ratio of 1:1, and then routinely embedded.

[0064] X4. Paraffin section

[0065] Cut the sample obtained by X3 into thin slices with a thickness of 3-6 μm.

[0066] X5. Baking Sheets and Baking Sheets

[0067] Place the thin slices with a thickness of 3-6 μm obtained in X4 in the systems of 37°C and 65°C for 8-12 hours respectively.

[006...

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Abstract

The invention discloses an embryo implantation detection kit, and application and a use method thereof. The invention discloses application of a uterine decidua NK cell Siglec-7 as a detection index in an embryo detection kit. The uterine decidua NK cell Siglec-7 in a uterine decidua sample is detected as the detection index. The kit comprises a buffer solution, a fixing agent, a dehydrating agent, an embedding agent, a dewaxing agent, an antigen repairing agent, a fluorescent reagent, a primary antibody, a secondary antibody, a redyeing solution and a mounting medium. The detection tool provided by the invention is used for detecting the embryo implantation capability, and the endometrial embryo implantation capability can be further evaluated. A possible detection and clinical diagnosismode is provided for clinical unexplained spontaneous abortion female patients, and a powerful reference is provided for discussing unrevealed influence factors of the unexplained spontaneous abortionfemale patients.

Description

technical field [0001] The invention relates to a kit and application thereof, in particular to an embryo implantation detection kit using uterine decidua NK cell Siglec-7 as a detection index and its application and usage method. Background technique [0002] The incidence of recurrent spontaneous abortion is 1%-5% in pregnancy, and the etiology is complex. In addition to chromosomal abnormalities, endocrine diseases, reproductive tract abnormalities and other reasons, there are still 70% of recurrent spontaneous abortions with unknown causes. Routine imaging and morphological tests cannot reveal the functional status of the endometrium. [0003] The State Intellectual Property Office published the application number CN201710649484.6 on September 22, 2017, and the title is an invention patent for a method and kit for preimplantation chromosome abnormality detection. Specifically, by improving single-cell expansion The augmentation method optimizes the steps of traditional...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6439
Inventor 马芳冯颖林羿母得志
Owner THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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