A kind of antigen for porcine infectious pleuropneumonia antibody detection and preparation method
A pleuropneumonia and antibody detection technology, applied in the direction of color/spectral characteristic measurement, measurement device, instrument, etc., can solve the problems of inability to meet the needs of clinical detection, lack of cross-protection, difficult to implement, etc., and achieve good detection effect, strong specificity, good stability
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[0014] The preparation method of the antigen of the present invention is as follows:
[0015] 1 Preparation of seeds for antigen production
[0016] 1.1 Preparation of chicken broth agar (referred to as S agar) Take 100ml of chicken broth, 3g of tryptone blood agar matrix, 0.5g of polypeptone, heat and dissolve, adjust the pH value to 7.2-7.4, sterilize at 115°C for 30 minutes, and cool to 55°C At about ℃, add 5ml of inactivated chicken serum and 1ml of 1% coenzyme I (NADH) according to the sterile requirements.
[0017] 1.2 Primary seed propagation and identification Streak inoculation of APP1-15 freeze-dried strains on S agar plate, place in 5%-10% CO 2 Under the conditions, culture at 37°C for 16-18 hours, select more than 5 typical colonies (or bacterial lawns) with iridescent luster, inoculate them into the yolk sac of SPF chicken embryos aged 5-6 days, continue to incubate at 37°C, and collect them within 30 hours The yolk fluid of dead chicken embryos shall be regarde...
Embodiment 1
[0037] 1 Preparation of seeds for antigen production
[0038] 1.1 First-level seed propagation and identification Streak inoculation of APP1-15 freeze-dried strains on S agar plate, place in 5%-10% CO 2 Under the conditions, culture at 37°C for 16-18 hours, select more than 5 typical colonies (or bacterial lawns) with iridescent luster, inoculate them into the yolk sac of SPF chicken embryos aged 5-6 days, continue to incubate at 37°C, and collect them within 30 hours The yolk fluid of dead chicken embryos shall be regarded as first-class seeds after passing the pure inspection. Stored at -20°C, the usable period should not exceed 1 month; stored below -70°C, the usable period should not exceed 3 months, and the subculture on the medium should not exceed 6 generations.
[0039] 1.2 Propagation of secondary seeds Take primary seeds of various types of strains, inoculate S agar plate by streaking, and inoculate with 5% to 10% CO 2 Under the conditions, cultivate at 37°C for 16...
Embodiment 2
[0051] 1 Preparation of seeds for antigen production
[0052] 1.1 First-level seed propagation and identification Streak inoculation of APP1-15 freeze-dried strains on S agar plate, place in 5%-10% CO 2 Under the conditions, culture at 37°C for 16-18 hours, select more than 5 typical colonies (or bacterial lawns) with iridescent luster, inoculate them into the yolk sac of SPF chicken embryos aged 5-6 days, continue to incubate at 37°C, and collect them within 30 hours The yolk fluid of dead chicken embryos shall be regarded as first-class seeds after passing the pure inspection. Stored at -20°C, the usable period should not exceed 1 month; stored below -70°C, the usable period should not exceed 3 months, and the subculture on the medium should not exceed 6 generations.
[0053] 1.2 Propagation of secondary seeds Take primary seeds of various types of strains, inoculate S agar plate by streaking, and inoculate with 5% to 10% CO 2 Under the conditions, cultivate at 37°C for 16...
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