HCV antigen detection board and method for detecting HCV antigen by using same
An antigen detection and detection board technology is applied in the detection of hepatitis C virus HCV antigen and the detection of virus antigens, which can solve the problems of low detection rate of hepatitis C virus core antigen, and achieve shortened detection time, high sensitivity, The effect of reducing the background
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Embodiment 1
[0048] Obtaining the HCV antigen detection panel:
[0049] A, preparation of anti-HCV serum
[0050] A1. Recombinant HCV antigen NS expressed in four highly conserved region genes with a concentration of 512Eu / ml 3 、NS 4 、NS 5 2 ml of the equal volume mixture of C and C, and immunize monkeys, rabbits or other suitable animals;
[0051] A2. Blood was collected at 2, 3, and 4 weeks respectively, and the HCV antibody titer was tested with a conventional hepatitis C virus antibody diagnostic kit. When the titer did not reach 1:4000, a booster immunization injection was required;
[0052] A3. When the anti-HCV serum titer is greater than 1:8000, blood is collected from the carotid artery, and serum is prepared according to a conventional method to obtain anti-HCV serum, which is stored below -60°C until use;
[0053] B. Purification of Anti-HCV Antibody
[0054] B1. Take protein A or protein A / G affinity chromatography column, equilibrate at room temperature for 30 minutes, an...
Embodiment 2
[0064] Obtaining the HCV antigen detection panel:
[0065] A, preparation of anti-HCV serum
[0066] A1. Recombinant HCV antigen NS expressed in four highly conserved region genes with a concentration of 1024Eu / ml 3 、NS 4 、NS 5 1ml of the equal volume mixture of C and C, to immunize monkeys, rabbits or other suitable animals;
[0067] A2. Blood was collected at 2, 3, and 4 weeks respectively, and the HCV antibody titer was tested with a conventional hepatitis C virus antibody diagnostic kit. When the titer did not reach 1:4000, a booster immunization injection was required;
[0068] A3. When the anti-HCV serum titer is greater than 1:8000, blood is collected from the carotid artery, and serum is prepared according to a conventional method to obtain anti-HCV serum, which is stored below -60°C until use;
[0069] B. Purification of Anti-HCV Antibody
[0070] B1. Take protein A or protein A / G affinity chromatography column, equilibrate at room temperature for 30 minutes, and...
Embodiment 3
[0089] Preparation of biotin-labeled HCV antibody use solution: After dissolving conventional long-arm biotin in water for injection as usual, mix it with conventional HCV monoclonal antibody at a molar ratio of 1:10; leave it at room temperature for 30 minutes, or After 2 hours in the refrigerator at 4°C, dialyze overnight with phosphate buffered saline PBS as usual, add an equal amount of glycerol, and store at -20°C for later use to obtain biotin-labeled HCV antibodies; use phosphate buffered saline PBS+2% volume ratio Dilute the biotin-labeled HCV antibody to a volume concentration of 1:500 to 1:2000 in the enzyme junction solution composed of bovine serum albumin to obtain a biotin-labeled HCV antibody use solution, which is stored at 4°C for future use.
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