Porcine circovirus 3 type antibody detection kit and application thereof
A porcine circovirus and antibody detection technology is applied in the biological field to achieve rapid detection, easy diagnosis and prevention and control
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[0017] As an embodiment of the present invention, in the ELISA antibody detection kit, the porcine circovirus type 3 Cap protein is the protein encoded by the sequence SEQ ID No.1.
[0018] The ELISA antibody detection kit of the present invention can be used to detect the antibody level in serum, and corresponding preventive measures can be provided accordingly to provide effective protection for pig farms, especially small pig farms.
[0019] As a preferred embodiment of the present invention, in the ELISA antibody detection kit, the support medium coated with porcine circovirus type 3 Cap protein is a microwell plate, and the porcine circovirus type 3 Cap protein is The coating concentration is 2-4 μg / ml, and the coating amount is 200-400 ng / well.
[0020] As an embodiment of the present invention, in the ELISA antibody detection kit, the kit further includes a sample diluent, a washing solution, a stop solution, a positive control, and a negative control.
[0021] The kit...
Embodiment 1
[0043] The preparation of embodiment 1 porcine circovirus type 3 Cap protein
[0044] 1.1 Isolation and identification of porcine circovirus type 3
[0045] Source of the disease: In a commercial pig farm in China, compared with the historical average, the mortality rate of sows increased by 9.4%, the pregnancy rate decreased by 1.2%, and the number of mummies increased by 8.2%. Clinically, affected sows are anorexic with signs of multifocal papules, macules and superficial dermatitis. Aborted litters contained mummified fetuses of different gestational ages, consistent with symptoms of PCV2 infection causing abortion. Although the overall clinical presentation and signs of abortion observed in sows were consistent with reproductive failure caused by PCV2, different tissues of all sows, including kidney, lymph nodes, lungs, skin and stillbirth, were detected by immunohistochemistry and quantitative PCR. Tests for PCV2, PRRSV, PPV, CSFV, and Mycoplasma hyopneumoniae were all ...
Embodiment 2
[0054] The preparation of embodiment 2 porcine circovirus type 3 ELISA antibody detection kit
[0055] 2.1 Preparation of ELISA antibody detection kit
[0056] ELISA antibody detection kit (referred to as kit 1) includes the following components:
[0057] Coated microwell plate: the porcine circovirus type 3 Cap protein prepared in Example 1 was coated with a coating concentration of 2-4 μg / ml, a coating amount of 200-400 ng / well, and 100 μl with pH9.6 carbonate buffer solution. Coat each well on a microplate, overnight at 2-8°C or for 2 hours at 37°C; discard the coating solution, wash 2-4 times with phosphate buffer, pat dry or drain; add blocking solution ( Such as 1-5% bovine serum albumin (BSA, 5% skimmed milk, 5% goat serum, 5% rabbit serum, 1% gelatin or 1% casein) at 37°C for 1.5-2 hours; discard the blocking solution and use phosphoric acid Wash 3 times with saline buffer and pat or drain dry.
[0058] Sample diluent: phosphate buffered saline.
[0059] Washing so...
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