Transfer factor against swine blue ear disease
A transfer factor and porcine blue-ear disease technology, which is applied in the field of transfer factor against porcine blue-ear disease, can solve the problems of weak drug competition, limited curative effect, and inability to prevent and reduce repeated freezing and thawing time. Accurate curative effect and long-lasting effect
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Embodiment 1
[0042] Embodiment 1: Preparation of anti-pig PRRS transfer factor
[0043] 1. Inoculate porcine PRRS virus VR2332 (Genbank accession No. 217415) or LV (Genbank accession No. 96262) in the allantoic fluid of 9-11-day-old chicken embryos and culture at 37°C for 48 hours. Take chicken embryo allantoic fluid, freeze and thaw repeatedly 5 times at -20°C, centrifuge at 5000rpm for 30min, discard the precipitate and collect the supernatant;
[0044] 2. Take the supernatant, and use the sucrose density gradient centrifugation to purify the supernatant to extract the virus. The sucrose concentration is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, and centrifuged at 150000rpm for 4 hours , determine the position of the centrifuge tube where the porcine PRRS virus is located according to the molecular weight of the porcine PRRS virus, and suck it out for subsequent use;
[0045] 3. The pigs immunized with the obtained porcine PRRS virus antigen were selected as healthy immunized pigs, and th...
Embodiment 2
[0055] Example 2: Preparation of anti-PRRS transfer factor
[0056] 1. Inoculate porcine PRRS virus in the allantoic fluid of 9-11 day old chicken embryos and culture at 37°C for 72h. Take chicken embryo allantoic fluid, freeze and thaw repeatedly 5 times at -20°C, centrifuge at 5000rpm for 30min, discard the precipitate and collect the supernatant;
[0057] 2. Get the supernatant, and the supernatant is purified by sucrose density gradient centrifugation to extract the virus. The sucrose concentration is 15%, 30%, 45%, 60%, 75%, centrifuged at 180000rpm for 3 hours, and determined according to the molecular weight of porcine PRRS virus The position of the centrifuge tube where the porcine PRRS virus is located is sucked out for later use;
[0058] 3. The pigs immunized with the obtained porcine PRRS virus antigen were selected as healthy immunized pigs, and the above-mentioned extracted virus antigens were subcutaneously injected into the immunized pigs at multiple points, a...
Embodiment 3
[0068] Example 3: Detection of the anti-PRRS transfer factor prepared by the process of the present invention
[0069] The anti-pig PRRS transfer factor prepared by the process described in Examples 1 and 2 is detected as follows:
[0070] 1. Ultraviolet spectrophotometric measurement: the sample obtained above has an absorption peak at 252±2nm, and ABS 260 / ABS 280 >2.0.
[0071] 2. The standard solution is light yellow and the pH value is between 6.0 and 8.0.
[0072] 3. Bacteriological examination: there is no need for aerobic, anaerobic, saprophytic bacteria and fungi in the samples obtained above.
[0073] 4. Detection of 20% sulfosalicylic acid: the sample obtained above has no turbidity and precipitation, indicating that the protein reaction is negative, and it does not contain macromolecular protein.
[0074] 5. Determination of nucleic acid content: the nucleic acid content of the samples obtained above was determined by the orcinol method to be 615.08 μg / mL and 5...
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