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Application of JUNO protein based on membrane block in rapid non-invasive detection of in vitro fertilization

A detection body and protein technology, applied in the field of reproduction, can solve the problems of fragile, invisible, and poor ICSI remedial effect.

Active Publication Date: 2021-07-20
WUHAN UNIV
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Problems solved by technology

However, clinical observations have found that the polar body of the oocyte is easily fragmented after being discharged, so it cannot be observed, or the first polar body splits into two second polar bodies, or the fragmentation into two parts easily leads to misjudgment
In prolonged fertilization methods, if fertilization fails, it will not be remedied because the eggs are already aged after 17-19 hours ICSI remedial effect is very poor

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

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

[0019] 1. Collect the fertilized droplets of a single oocyte and sperm from a female patient 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 were tested by ELISA to detect the negative or positive of JUNO protein in the fertilized droplets.

[0022] (2) Specific steps include:

[0023] 1. Coating: Dilute the fertilized droplets after centrifugation to 100ul with a buffer (carbonate buffer of 0.05MPH9.6), 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, pH7.5, the same below) for 3 times, each time for 2 minutes; add 100 μl of blocking solution (containing 3% BSA PBST), blocked at 37°C for 60 min; washed 3 times with PBST.

[0...

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Abstract

The present invention provides an application of the JUNO protein based on the membrane block in rapid non-invasive detection of in vitro fertilization. By detecting the JUNO protein released by the membrane block when oocytes are fertilized in fertilization droplets, it is possible to quickly and early judge whether the fertilization is successful. The present invention is based on the detection of the specificity of the JUNO protein released by the membrnae 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 the JUNO 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 earlier. If the fertilization is not successful, the oocytes have not yet aged at this time, and then quickly determine whether ICSI is needed for remediation. The detection method is non-invasive. It is suitable for large-scale promotion and application.

Description

technical field [0001] The invention belongs to the technical field of reproduction, and in particular relates to the application of a membrane block-based JUNO protein in rapid non-invasive detection of in vitro fertilization. Background technique [0002] In the field of assisted reproduction, in vitro fertilization technology has brought good news to the majority of infertility patients. Although the rate of in vitro fertilization is between 60% and 80%, there are still some cases where complete fertilization fails or the fertilization rate is low. At present, due to the limited detection methods, it is difficult to accurately and timely detect whether oocyte fertilization is successful, resulting in waste of resources, bringing great psychological pressure and pain to patients, and also causing huge human and financial resources to patients waste. Therefore, how to timely and accurately detect the success of in vitro fertilization has become a research and exploration ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/53
CPCG01N33/53G01N33/68
Inventor 罗孟成刘聪吴月蓉
Owner WUHAN UNIV
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