Detection kit and detection method for porcine circovirus type 2 antibody
A porcine circovirus and detection kit technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of long detection time, difficulty in quantification, and difficulty in automation, and achieve short reaction time, improved repeatability, and wide detection range Effect
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Embodiment 1
[0098] A kit for detecting porcine circovirus type 2 antibody, comprising magnetic suspension coated with porcine circovirus type 2 Cap protein, alkaline phosphatase-labeled porcine circovirus type 2 antibody solution, diluent, calibrator, Quality control products, washing solutions, luminescent substrates.
[0099] Preparation of magnetic suspension coated with porcine circovirus type 2 Cap protein:
[0100] (1) Take 1 mL of magnetic particles (purchased from JSR Corporation, Japan, item number: Magnosphere TM MS300 / Caboxyl) solution with a concentration of 10 mg / mL, washed twice with 0.1 mol / L 2-morpholineethanesulfonic acid (MES) buffer solution with a pH of 5.0, and finally suspended in 1 mL of 0.1 mol / L pH 5.0 In MES buffer;
[0101] (2) 70 μg of purified porcine circovirus type 2 Cap protein (purchased from Qingdao Yibang Bioengineering Co., Ltd.) was added;
[0102] (3) Weigh 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), dissolve it with 0.1mol / L MES buffer s...
Embodiment 2
[0164] A kit for porcine circovirus type 2 antibody detection, comprising a magnetic suspension coated with porcine circovirus type 2 Cap protein, and acridinium ester-labeled goat anti-pig IgG antibody (purchased from Beijing Suo Laibao Technology Co., Ltd.) Solutions, diluents, calibrators, quality controls, wash solutions, primary luminescent substrates and secondary luminescent substrates.
[0165] Preparation of magnetic suspension coated with porcine circovirus type 2 Cap protein:
[0166] (1) Take 1mL of the solution containing magnetic particles with a concentration of 10mg / mL, wash it twice with 0.1mol / L boric acid buffer solution with a pH of 9.5, and finally suspend in 1mL of a 0.1mol / L boric acid buffer solution with a pH of 9.5;
[0167] (2) Add 80 μg of purified porcine circovirus type 2 Cap antigen, vortex and mix;
[0168] (3) Add 0.5 mL of 0.1 mol / L boric acid buffer (containing 3 mol / L ammonium sulfate) with a pH of 9.5, and shake and react at 37°C for 20 ho...
Embodiment 3
[0212] A test kit for porcine circovirus type 2 antibody detection, comprising a magnetic suspension coated with porcine circovirus type 2 Cap protein, horseradish peroxidase-labeled porcine circovirus type 2 Cap antigen solution, diluent, Calibrator, Quality Control, Washing Solution, Primary Luminescent Substrate and Second Luminescent Substrate.
[0213] Preparation of magnetic suspension coated with porcine circovirus type 2 Cap protein:
[0214] (1) Take 1mL of the solution containing magnetic particles, the concentration is 10mg / mL, wash twice with 0.01mol / L phosphate (PBS) buffer solution with pH 7.4, and finally suspend in 1mL 0.01mol / L PBS with pH 7.4 in the buffer;
[0215] (2) Add 0.1 mL of 25% (v / v) glutaraldehyde solution, shake and react at 37°C for 2 hours;
[0216] (3) Wash 3 times with 1mL 0.01mol / L PBS buffer solution with pH 7.4;
[0217] (4) Adding 30 μg of purified porcine circovirus type 2 Cap recombinant antigen, shaking and reacting at 37°C for 20 ho...
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