Application of zona block-based ovastacin protein in rapid non-invasive detection of in vitro fertilization

A technology for detecting bodies and proteins, applied in the field of reproduction, can solve the problems of unobservation, misjudgment as polar bodies, and many shortcomings in the observation of second polar bodies.

Active Publication Date: 2021-06-15
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In prolonged fertilization methods, failure to fertilize will not be remedied as eggs are already aged after 17-19 hours ICSI remediation is poor
There are many shortcomings in the observation of the second polar body in the short-term fertilization method, which may easily lead to misjudgment
For example: after the polar body is discharged, it is easy to break and cannot be observed; theoretically, the first polar body may be split into two polar bodies, or broken into two parts, which will be misjudged as a first polar body and a second polar body Diodes, which are misjudged as successful fertilization or egg removal, or other fragments similar to polar bodies may be produced in the zona pellucida during the culture process, which will be misjudged as polar bodies, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] (1) The collection and detection process of ELISA detection ovastacin protein is as follows:

[0019] 1. Collect the fertilized droplets of the female patient's single egg and sperm after 4-6 hours of co-cultivation.

[0020] 2. Centrifuge at 3000rpm at 4°C for 5 minutes to remove excess sperm.

[0021] 3. The fertilized droplets are subjected to ELISA detection to detect the negative or positive of ovastacin protein in the fertilized droplets.

[0022] (2) Specific steps include:

[0023] 1. Coating: Dilute the fertilized droplets after centrifugation to 100ul with a buffer (0.05M PH 9.6 carbonate buffer), and coat a 96-well ELISA microtiter plate (Corning company product, the same below), 37 ℃, coating for 1h;

[0024] 2. Blocking: wash the coated ELISA plate with PBS-T (PBS containing 0.1% Tween-20, pH 7.5, the same below) for 3 times, 2 min each time; add 100 μl blocking solution (PBST containing 3% BSA ), blocked for 60 min at 37°C; washed three times with PBST...

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Abstract

The present invention provides an application of zona block-based ovastacin protein in rapid non-invasive detection of in vitro fertilization, by detecting the ovastacin protein released from oocyte zona block in fertilized droplets, early and accurately judging whether the fertilization is successful or not. The present invention is based on the detection of the specificity of the ovastacin protein released by the zona block mechanism of the oocyte during the fertilization process. The protein is released from the oocyte only after the oocyte is successfully fertilized to prevent polyspermy. By detecting ovastacin protein to determine whether the oocyte was fertilized successfully. Compared with the traditional method of egg removal by scientific researchers, this method is more objective and the detection time is relatively early. At this time, the oocytes have not yet aged, and then quickly determine whether ICSI is needed for remediation. The detection method is non-invasive and suitable for large-scale Promote apps.

Description

technical field [0001] The invention belongs to the technical field of reproduction, and in particular relates to the application of zona block-based ovastacin protein in rapid non-invasive detection of in vitro fertilization. Background technique [0002] In vitro fertilization-embryo transfer technology uses drugs to make a batch of women's eggs grow and mature synchronously. After being taken out by puncture, they are placed in a petri dish with her husband's optimized sperm for co-incubation. After a period of in vitro fertilization, they develop into fertilized eggs. , and then gradually develop into embryos by cleavage; then select high-quality embryos and place them in the woman's uterus, and finally develop into fetuses. [0003] There are currently two main methods of in vitro fertilization in assisted reproduction, namely long-term fertilization and short-term fertilization. Long-term fertilization is to judge whether the fertilization is successful by observing t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573G01N33/569
CPCG01N33/56966G01N33/573G01N2333/96491G01N2800/36
Inventor 罗孟成刘聪吴月蓉
Owner WUHAN UNIV
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