Chemiluminescence immunoassay kit for Seneca valley virus nonstructural protein 3ABC antibody detection
A chemiluminescence immunoassay, non-structural protein technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, material analysis by optical means, etc., to achieve high protein yield, simple operation, and improved detection sensitivity. Effect
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Embodiment 1
[0049] Example 1 Preparation and assembly of chemiluminescent immunoassay kit for detection of Seneca Valley virus nonstructural protein 3ABC antibody
[0050] (1) Preparation of the kit:
[0051] 1. Expression of Seneca Valley virus nonstructural protein 3ABC antigen:
[0052] The 969bp gene encoding the Seneca Valley virus nonstructural protein 3ABC (shown in SEQ ID NO.1) was cloned into the pET-30a(+) prokaryotic expression vector. After digestion and sequencing were correct, the positive plasmid was transformed into the BL21DE3plysS strain, Kanamycin-resistant LB plates were used to screen single clones, shake the LB medium at 37°C overnight; transfer the overnight bacterial solution to fresh LB medium at a volume ratio (v / v) of 1:100, shake at 37°C at 200rpm After 3 hours until the OD600 value reaches about 0.6, add IPTG with a final concentration of 1 mmol / L to continue shaking the bacteria for 3 hours, and collect the bacterial liquid by centrifugation at 6000 rpm. Re...
Embodiment 2
[0089] Example 2 Sensitivity and specificity experiments of the chemiluminescent immunoassay kit for detection of Seneca Valley virus nonstructural protein 3ABC antibody
[0090] 1. Dilution of known background serum:
[0091] (1) A total of 30 serum samples were obtained from clinically healthy pigs that had not been infected with Seneca Valley virus. These sera were used to evaluate the specificity of the chemiluminescent detection method for antibodies to nonstructural proteins of Seneca Valley virus.
[0092] (2) 30 samples each of porcine vesicular disease positive serum, classical swine fever virus positive serum, and porcine circovirus-2 positive serum, these sera were used to evaluate the specificity of the chemiluminescent detection method for Seneca Valley virus nonstructural protein 3ABC antibody .
[0093] (3) A total of 80 serum samples were from pigs challenged experimentally with SVV. The pigs had not received any immunizations prior to the challenge and these...
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