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Protein biomarker for diagnosis of early pregnancy of sheep and method for detecting early pregnancy of sheep by using protein biomarker

A technology of biomarkers and protein markers, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as unclear symptom diagnosis, influence of experience level, long detection time and days, and achieve high early pregnancy detection value Effect

Active Publication Date: 2021-08-27
XINJIANG ACADEMY OF AGRI & RECLAMATION SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Traditional methods of pregnancy diagnosis, such as testing, external observation, ultrasonic diagnosis, vaginal examination, etc., have their own defects, the detection time is long, the symptom diagnosis is not clear, and it is affected by the experience of the user, etc. Misdiagnosis, There are varying degrees of misdiagnosis

Method used

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  • Protein biomarker for diagnosis of early pregnancy of sheep and method for detecting early pregnancy of sheep by using protein biomarker
  • Protein biomarker for diagnosis of early pregnancy of sheep and method for detecting early pregnancy of sheep by using protein biomarker
  • Protein biomarker for diagnosis of early pregnancy of sheep and method for detecting early pregnancy of sheep by using protein biomarker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037]Example 1: Screening of differential proteins in the peripheral blood of pregnant and non-pregnant sheep on the 14th day after mating by TMT technology

[0038] In this example, the blood of pregnant and non-pregnant sheep was collected on the 14th day after mating, the proteins were separated, and the proteins differentially expressed in early pregnancy were analyzed and screened using TMT technology and bioinformatics methods. The specific steps are as follows:

[0039] 1. Test samples

[0040] Eight normal Kazakh sheep were randomly divided into control group and experimental group. According to the conventional production process, the ewes in the test group received normal semen, and the ewes in the control group received dead sperm (the normal semen was boiled at high temperature and observed under a microscope to ensure that the sperm died). On the 14th day after mating, the blood of pregnant ewes and ewes of the control group was collected, the plasma was separat...

Embodiment 2

[0088] Example 2: Large group verification of ELISA differential protein expression in pregnant and non-pregnant peripheral blood 14 days after mating In order to further verify the protein expression results of TMT detection, we used ELISA technology to detect the expression of four proteins related to early pregnancy in sheep The situation was verified with a large group.

[0089] The ELISA kits of sheep chitinase-3-like protein-1 (CHI3L1), sheep proteasome B subunit type 4 (PSMB4) and sheep soluble galectin 3 (LGALS3BP) were purchased from Shanghai Hengyuan Biotechnology Co., Ltd. .

[0090] 1. Sample information collection

[0091] Blood samples from 30 sheep were collected on the 14th day after mating, and the serum was separated, frozen and stored in dry ice immediately, and transported back to the laboratory as soon as possible and stored in an ultra-low temperature refrigerator for later use.

[0092] 2. Sample information confirmation

[0093] After the blood sampl...

Embodiment 3

[0134] Example 3: Application of protein biomarkers in the diagnosis of early pregnancy in sheep

[0135] This example provides the application of a protein biomarker in the diagnosis of early pregnancy in sheep. Whether the sheep is pregnant can be judged by detecting the expression level of protein biomarkers. In this case, ELISA enzyme-linked adsorption reaction is used as an example to detect. The ELISA kit was purchased from Shanghai Hengyuan Biotechnology Co., Ltd.

[0136] 1. Types and Collection of Blood Samples

[0137] Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or overnight at 4°C, then centrifuge at 1000×g for 20 minutes, take the supernatant, or store the supernatant at -20°C or -80°C , but repeated freezing and thawing should be avoided.

[0138] Plasma: Use EDTA or heparin as an anticoagulant to collect samples, and centrifuge the samples at 1000×g at 2-8°C for 15 minutes within 30 minutes after co...

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Abstract

The invention discloses a protein biomarker which is one of a CHI3L1 protein with the serial number being <210>1, a PSMB4 protein with the serial number being <210>2, and an LGALS3BP protein with the serial number being <210>3. The protein biomarker is applied to early pregnancy diagnosis of sheep. The invention discloses a sheep early pregnancy diagnosis method which is characterized by comprising the specific steps of (1) conducting blood sample collection, specifically, collecting sheep peripheral blood 14 days after hybridization; (2) carrying out blood sample treatment, specifically, carrying out serum separation or plasma treatment on the blood sample in the step (1); and (3) performing kit detection, specifically, detecting the blood sample treated in the step (2) by using an ELISA kit, and detecting the contents of the CHI3L1 protein, the PSMB4 protein and the LGALS3BP protein.

Description

technical field [0001] The invention belongs to the field of biotechnology diagnosis, and in particular relates to a protein biomarker used for the diagnosis of early sheep pregnancy and a method for the diagnosis of early sheep pregnancy. Background technique [0002] The pregnancy diagnosis of sheep has always been valued by enterprises, because the accurate diagnosis of early pregnancy can not only reduce the economic loss caused by empty pregnant ewes, but also facilitate timely grouping of pregnant ewes, reasonable feeding and management, and also have It is beneficial to improve the breeding efficiency of sheep, promote the intensive management of livestock, and improve the efficiency. How to find non-pregnant ewes, shorten the parity interval and reduce the number of non-productive days (NPD) through effective and accurate diagnostic methods has been perplexing the sheep industry. [0003] Traditional methods of pregnancy diagnosis, such as testing, external observat...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/689
Inventor 黄涛王衠萱公红斌孙义姗杨敏杨华徐梦思谢苏高若男孙晓梅和军飞卢世豪李清春陈鑫祁梦凡黎明国何凡
Owner XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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