A kind of porcine blue ear virus chimeric antigen and colloidal gold immunochromatographic test strip for detecting porcine blue ear virus antibody
An immunochromatographic test strip, porcine blue ear virus technology, applied in the direction of virus/phage, virus, virus peptide, etc., can solve the problems of missed detection, easy to disperse virus, etc., achieve high specificity, high practical value, easy to promote and use Effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0027] Example 1: Recombinant expression of the chimeric antigen of the present invention
[0028] 1. Construction of recombinant expression plasmid pGAPZ-NG for porcine PRRS virus chimeric antigen gene
[0029] Using bioinformatics technology to analyze the antigenicity and conservation of the structural proteins N protein and GP5 protein of different lineage strains of porcine PRRS virus, design and synthesize the specific chimeric antigen NG gene of porcine PRRS virus and optimize it, and use chemically synthesized Methods The chimeric antigen gene sequence of porcine PRRS virus (shown in SEQ ID NO: 2) was synthesized. Digest the pGAPZ vector with HindⅢ and NotI. The digestion system is 50uL: 1uL each of HindⅢ and NotI, 20uL of the vector fragment, 5uL of 10X digestion buffer, and 23uL of ddH2O. box recycling. Then the digested vector was ligated with the NG gene sequence to construct a 10 μL ligation system: 5.5 μL of the NG gene fragment, 2.5 μL of the pGAPZ vector, 0.5...
Embodiment 2
[0039] Embodiment 2: Colloidal gold immunochromatographic detection test strip for preparing porcine blue ear virus antibody
[0040] 1. Preparation of gold-labeled conjugates of SPA labeled with gold particles
[0041] Take 2ml of 40nm colloidal gold particles and use 0.1mol / L potassium carbonate to adjust the pH to 6.8, add 10ug Staphylococcus aureus protein A (SPA), mix quickly and mix at room temperature on a 3D rotator for 30min, then add the final concentration 1% BSA and mix on a 3D rotary mixer for 30min, centrifuge the gold standard solution at 12000r / min at 4°C for 10min, carefully discard the supernatant, wash the precipitate twice with 0.01M PBS buffer, and then centrifuge the resulting precipitate It is the purified SPA gold-labeled complex. The prepared colloidal gold-labeled SPA is resuspended in 0.01MPBS and stored at 4°C for later use.
[0042] 2. Nitrocellulose membrane testing line and quality control line spraying and colloidal gold bonding pad preparation...
Embodiment 3
[0048] Embodiment 3: Colloidal gold immunochromatography test strip specificity test
[0049] Crossover test was carried out on the known porcine foot-and-mouth disease type O positive serum, porcine circovirus type II positive serum, porcine rotavirus positive serum, classical swine fever virus positive serum and porcine blue ear virus positive serum, the results are shown in figure 2 .
[0050] figure 2 The results showed that only the porcine blue ear virus positive serum sample had a positive reaction with the detection line, but when it reacted with several other sera, the detection line had no color development and a negative reaction, indicating that the chimeric antigen was used for detection Antibodies against porcine PRRS virus are highly specific.
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More