Swine Seneca valley virus high-titer positive serum and preparation method thereof
A swine Seneca, positive serum technology, applied in biochemical equipment and methods, viruses, viruses/phages, etc., can solve the problem that there is no porcine Seneca Valley virus hyperimmune serum, no porcine Seneca Valley virus specificity. In order to reduce the risk of exogenous viruses, provide a large amount of serum, and produce high antigen yields
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[0032] The invention provides a method for preparing porcine Seneca Valley virus-specific high-titer positive serum, using guinea pigs as immunized animals, using the virus liquid of suspended cultured cells as porcine Seneca Valley virus antigen, and adopting porcine Seneca Valley virus antigen The guinea pigs were stimulated by the synergistic action of the solution and the inactivated vaccine to obtain positive sera with high and high titers.
[0033] The test animal that the present invention adopts is 350~450g guinea pig, and described guinea pig is not higher than 1:4 to porcine Seneca Valley virus serum neutralizing antibody titer, porcine circovirus type 2 antibody, porcine blue ear virus antibody detection Negative, swine Seneca Valley virus, foot-and-mouth disease virus, African swine fever virus antigen tests were negative.
[0034] The invention provides a method for preparing porcine Seneca Valley virus-specific high-titer positive serum, comprising the following ...
Embodiment 1
[0070] Example 1 The preparation method of porcine Seneca Valley virus-specific high-titer positive serum
[0071] 1. Preparation of porcine Seneca Valley virus liquid
[0072] After the cells were revived, they were cultured in suspension at 37±0.5°C in a shaker, and the cell density reached 4×10 6 When the cells / mL is above, scale up the culture step by step. Cell density up to 4×10 6 cells / mL, and when the cell viability is above 90%, inoculate the cultured suspension cells with an amount of 0.1% of the volume of the cell fluid to produce the virus seed, and set the culture parameters (pH value, DO value, stirring speed) at 37 Cultivate at ±0.5°C, take samples for 24 hours, observe under a microscope, harvest the virus culture when the cell viability is less than 20%, and freeze and thaw once (the purpose of freezing and thawing: to place the harvested virus liquid at -18°C overnight, Then thaw at room temperature to completely release the virus from the cells), and stor...
Embodiment 2
[0083] Example 2 The preparation method of porcine Seneca Valley virus-specific high-titer positive serum
[0084] 1. Preparation of porcine Seneca Valley virus antigen solution
[0085] After the cells were revived, they were cultured in suspension at 37±0.5°C in a shaker, and the cell density reached 4×10 6 When the cells / mL is above, scale up the culture step by step (pH7.2±0.1, dissolved oxygen DO 30%-60%, stirring speed 60-150rpm). Cell density up to 4×10 6cells / mL, and when the cell viability is above 90%, inoculate the cultured suspension cells with the amount of 0.5% of the cell fluid volume of the production virus seed, and set the culture parameters (pH7.2±0.1, dissolved oxygen DO 40%~ 60%, stirring speed 80~110rpm) cultivated at 37±0.5°C, sampled in 24 hours, observed under a microscope, when the cell viability was less than 20%, the virus culture was harvested, freeze-thawed once (freeze-thaw purpose: to harvest the The virus liquid was left standing overnight a...
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Abstract
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