Diagnosis marker kallistatin for ovarian hyperstimulation syndrome and its application

A marker and syndrome technology, applied in the field of biotechnology and medical testing, can solve problems such as insufficient ovarian hyperstimulation syndrome

Active Publication Date: 2021-03-16
GUANGDONG NO 2 PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to ultrasound technology, most of the existing technologies use its clinical manifestations and basic information to support the disease, and there is no sufficient evidence and disease mechanism to support the occurrence of ovarian hyperstimulation syndrome.

Method used

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  • Diagnosis marker kallistatin for ovarian hyperstimulation syndrome and its application
  • Diagnosis marker kallistatin for ovarian hyperstimulation syndrome and its application
  • Diagnosis marker kallistatin for ovarian hyperstimulation syndrome and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1Kallistatin protein is widely expressed in ovarian granulosa cells

[0032] This example detects the expression of Kallistatin protein in granulosa cell line KGN and primary ovarian granulosa cells.

[0033]1. Expression of Kallistatin in granulosa cell line KGN cells

[0034] The granulosa cell line KGN cells were cultured, and the expression of Kallistatin factor in the cells was detected by immunofluorescence method.

[0035] (1) Culture of KGN cells

[0036] Take DMEM / F12 medium, add FBS (10%) and penicillin / streptomycin double antibody (1%) to prepare cell culture medium, culture cells in a 100mm cell culture dish, and change the medium every other day until the cell density reaches 80% Cells were passaged at left and right.

[0037] (2) Cell immunofluorescence detection

[0038] 1) Digest and centrifuge the cells with a degree of confluence of about 70%, resuspend with 3ml of cell culture medium, add about 250ul of cell suspension to a small dish of l...

Embodiment 2

[0063] Example 2 Expression of Kallistatin protein in clinical samples of high-risk OHSS and assisted reproduction control patients

[0064] This example studies the expression of Kallistatin protein in the follicular fluid and serum of OHSS high-risk group and control group patients undergoing assisted reproduction.

[0065] On the basis of the informed consent of the patients and the consent of the hospital ethics committee, patients were selected for in vitro fertilization / intracytoplasmic sperm injection-embryo transfer (IVF) in the reproductive center of the Third Affiliated Hospital of Guangzhou Medical University from March 2016 to August 2016. / ICSI-ET) assisted infertility female patients, collect follicular fluid and / or blood samples on the day of egg retrieval, and / or extract granulosa cells from the follicular fluid for experiments. Patients were divided into two groups: OHSS high-risk group and control group according to their general characteristics such as age, ...

Embodiment 3

[0104] Example 3 Expression of Kallistatin gene in clinical samples of high-risk OHSS and assisted reproduction control patients

[0105] This example studies the expression of Kallistatin gene in the follicular fluid of OHSS high-risk group and control group patients undergoing assisted reproduction.

[0106] 1. RNA extraction: Extract the total RNA from the follicular fluid of the control group patients with high risk of OHSS and assisted reproduction. Collect an appropriate amount of follicular fluid from the patient on the day of egg retrieval, centrifuge at 1500-1800rpm at 4°C for 15min, and pipette 1ml of supernatant into two 2ml EP tubes, and mark the patient's name, group and other information on the tube wall. Store in -80°C refrigerator. Add an appropriate amount of Trizol, repeatedly pipetting to lyse the cells, and then transfer into an RNase-free EP tube; if it is tissue, use a mortar to grind the tissue, then add an appropriate amount of Trizol, and repeatedly p...

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Abstract

The invention discloses an ovarian hyperstimulation syndrome (OHSS) detection marker Kallistatin, and also discloses an application of Kallistatin protein in screening and / or preparation of an ovarianhyperstimulation syndrome detection reagent, and the ovarian hyperstimulation syndrome detection kit. Through research, the inventor of the invention finds that Kallistatin is widely expressed in ovarian granular cells, and the expression level in cell nucleuses is higher than that in cytoplasm. The inventor finds for the first time that compared with a control group for assisted reproduction, inclinical follicular fluid specimens and serum specimens of an OHSS high-risk group, the expression of Kallistatin protein is obviously reduced; compared with a normal pregnant woman, the content of Kallistatin in serum of an OHSS patient is remarkably reduced. It is prompted that Kallistatin can be used as a specific detection marker of OHSS, is used for screening and preparing an OHSS detectionkit, and has good sensitivity and specificity. The invention provides a novel specific molecular marker for clinical detection of OHSS, and enriches the clinical detection method of OHSS.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and medical detection, in particular to Kallistatin, a diagnostic marker for ovarian hyperstimulation syndrome, and applications thereof. Background technique [0002] Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic disease related to assisted reproductive technology and a major complication of controlled ovarian hyperstimulation. It can seriously threaten the life of patients and should be actively prevented and treated. OHSS is characterized by the development of multiple follicles in bilateral ovaries, enlarged ovaries, abnormal capillary permeability, and extravasation of abnormal body fluids and proteins into the third space of the human body, which causes a series of clinical symptoms. It usually occurs in the first trimester after an HCG trigger or after transplantation. OHSS is often clinically manifested as nausea, vomiting, abdominal distension, oliguria, weight gain, an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68C12Q1/6883
CPCC12Q1/6883C12Q2600/158G01N33/6893G01N2333/8121G01N2800/36
Inventor 姚亚超李磊易婧雅刘静曹东林
Owner GUANGDONG NO 2 PROVINCIAL PEOPLES HOSPITAL
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