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Method for judging cell state in body liquid environment based on liquid-phase chip technology

A liquid phase chip, cell state technology, applied in the field of assisted reproduction, can solve the problem of no ovum/embryo quality-related factors, and achieve the effect of improving the clinical pregnancy rate and embryo implantation rate, and improving the treatment outcome.

Active Publication Date: 2014-04-09
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] At present, commercialized liquid-phase chip kits include inflammatory factor combinations, tumor factor combinations, diabetes combinations, etc., but there are no factors related to egg / embryo quality

Method used

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  • Method for judging cell state in body liquid environment based on liquid-phase chip technology
  • Method for judging cell state in body liquid environment based on liquid-phase chip technology
  • Method for judging cell state in body liquid environment based on liquid-phase chip technology

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Experimental program
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Effect test

Embodiment 1

[0035] 1. According to the literature, select factors that are related to the potential of egg development and can predict the successful implantation of embryos, such as granulocyte-colony stimulating factor (CSF), interleukin 15 (interleukin 15, IL-15), stem cell factor ( stem cell factor, SCF) and interferon-inducible protein 10.

[0036] 2. Select and purchase antibodies suitable for liquid chip technology for the selected factors

[0037] 3. Protein preparation: If the antibody does not contain sodium azide, BSA, glycine, Tris or other additives containing free amino groups, it can be used directly for coupling; otherwise, the buffer needs to be replaced with a Bio-spin mini-column.

[0038] 4. Coupling reaction:

[0039] a. Microsphere activation: select microspheres, vortex to mix, centrifuge to remove supernatant, add EDC (50 mg / ml) and S-NHS (50 mg / ml), vortex to mix, centrifuge to remove supernatant, add PBS to refill hanging.

[0040] b. Protein coupling: Add 5-1...

Embodiment 2

[0045] 1. The patient undergoes superovulation induction, and after the follicles mature, eggs are collected under the guidance of B-ultrasound. The contents of one follicle are collected into a sterile test tube and poured into a sterile petri dish, and the eggs are searched under a stereo microscope to find the eggs. Then transfer to G-IVF (Vitrolife) culture medium, cultivate in 37 degrees, 6% carbon dioxide incubator, and the corresponding culture medium is analyzed by liquid phase chip technology for biomarkers.

[0046] 2. The procedure for measuring follicular fluid biomarkers is as follows:

[0047] (1) Prepare the specimen and centrifuge at 1000g for 15 minutes

[0048] (2) Prepare the coupled microspheres: add an appropriate amount of buffer to a 15 ml test tube, vortex the coupled microspheres for 30 seconds, add an appropriate amount of microspheres to the 15 ml test tube, keep in the dark, and equilibrate at room temperature for 20 minutes.

[0049] (3) Add cou...

Embodiment 3

[0055] During 2011.10-2012.3, we selected some in vitro fertilization-embryo transfer patients to conduct follicular fluid liquid-phase chip biomarker detection. The preliminary results showed that increasing the liquid-phase chip biological objective indicators would help increase the embryo implantation rate. For example, if G-CSF is selected, the accuracy rate of embryo implantation prediction can reach 80.6%, while the embryo implantation rate is generally around 30%-40%. See results Figure 5 .

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Abstract

The invention provides a method for judging a cell state in a body liquid environment based on a liquid-phase chip technology. A biological marker of a follicular fluid is analyzed by using the liquid-phase chip technology, so as to judge the quality of forming an embryo by an ovum from the same follicular of the follicular fluid; good-quality embryo selection is determined by combining with a morphological analysis. With the adoption of the method provided by the invention, the clinical pregnancy rate and the embryo implantation rate of IVF (In-Vitro Fertilization) can be improved by providing an objective indicator of the ovum quality, so as to improve the result of IVF therapy. A plurality of factor combinations for judging the ovum quality of the follicular fluid can be produced into a commercial liquid-phase chip kit which is used for assisting the reproduction technology.

Description

technical field [0001] The invention belongs to the technical field of assisted reproduction, and relates to a method for judging the state of cells in a body fluid environment based on a liquid chip technology, which is a method for judging the quality of eggs by analyzing components in the body fluid based on the liquid chip technology. Background technique [0002] (1) In vitro fertilization technology [0003] In vitro fertilization means that mammalian sperm and eggs are cultured together in vitro and developed into fertilized eggs (also called embryos) under strict laboratory conditions, because the fertilization process of this technology is completed in an artificially controlled environment in vitro Therefore, it is called in vitro fertilization (in vitro fertilization, IVF). After fertilized eggs are obtained through IVF technology, the fertilized eggs are placed in the uterus to develop into fetuses. This process is called embryo transfer (ET). In vitro fertiliz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/68G01N33/74
CPCG01N33/54313G01N33/68
Inventor 叶英辉黄荷凤周承亮曲凡
Owner ZHEJIANG UNIV
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