Protein biomarker for early pregnancy diagnosis of sows and method for early pregnancy diagnosis of sows
A technology of biomarkers and protein markers, which is applied in biological testing, biomaterial analysis, instruments, etc., can solve problems such as long diagnosis time, influence of user experience, and unclear diagnosis of symptoms, so as to shorten the parity interval , Reduce the number of non-production days, reduce the effect of production costs
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Embodiment 1
[0047] Example 1: Screening of differential proteins in peripheral blood of pregnant and non-pregnant sows on the 15th day after mating by iTRAQ technology
[0048] In this example, the blood of pregnant and non-pregnant sows on the 15th day after mating was collected, the proteins were separated, and iTRAQ technology and bioinformatics methods were used to analyze and screen the differentially expressed proteins in the early pregnancy. The specific steps are as follows:
[0049] 1. Test sample
[0050] Eight normal-grown binary sows were randomly divided into control group and experimental group. According to the routine production process to check the estrus, the sows in the test group were injected with normal semen, and the sows in the control group were injected with dead semen (normal semen was boiled at high temperature and observed under a microscope to ensure that the sperm died). Blood was collected from pregnant sows and control sows on the 15th day after mating, a...
Embodiment 2
[0094] Example 2: Large group validation of differential protein expression in 15 days after breeding by ELISA between pregnant and non-pregnant peripheral blood
[0095] To further verify the protein expression results detected by iTRAQ, we used ELISA technology to verify the expression of the above four proteins in a large group.
[0096] ELISA kits for telomere repeat binding factor 2 (TERF2), haptoglobin (HP), adiponectin (ADIPOQ) and filamin C (FLNC) were purchased from Shanghai Hengyuan Biotechnology Co., Ltd.
[0097] 1. Sample information collection
[0098] On the 15th day after breeding, 21 sow blood samples were collected, and the plasma was separated, immediately placed in dry ice for cryopreservation, and transported back to the laboratory as soon as possible and stored in an ultra-low temperature refrigerator for future use.
[0099] 2. Sample information confirmation
[0100] After blood sample collection, B-ultrasound pregnancy test was performed on all breed...
Embodiment 3
[0142] Example 3: Application of protein biomarkers in the diagnosis of early pregnancy in sows
[0143] This example provides the application of a protein biomarker in the diagnosis of early pregnancy in sows. By detecting the expression level of protein biomarkers, we can judge whether the sow is pregnant. In this example, the ELISA enzyme-linked adsorption reaction was used as an example to diagnose early pregnancy. The ELISA kit was purchased from Shanghai Hengyuan Biotechnology Co., Ltd.
[0144] 1. Types and collection of blood samples
[0145] 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.
[0146] Plasma: collect specimens with EDTA or heparin as anticoagulant, centrifuge the specimens at 2-8°C at 1000×g for 15 minutes within ...
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