Vaccine for use in protecting a pig against porcine endemic diarrhea virus
一种猪流行性腹泻、疫苗的技术，应用在保护猪免于PEDV感染领域，能够解决未明确报告实际保护等问题
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Embodiment 1
[0029] Example 1 Virus neutralization assay
[0030] Sera were tested for the presence of neutralizing antibodies against PEDV using a virus neutralization (VN) assay on Vero cells. Briefly, serial two-fold dilutions of test samples or reference sera were prepared in aMEM-TPB cell culture medium containing the antibiotics penicillin and streptomycin. Each dilution with an equal volume of 100 TCID 50Infectious PEDV strains expressing green fluorescent protein (GFP) after infection of susceptible Vero cells were mixed. After pre-incubation for 1 h at 37 °C, the virus / serum dilution mixture was transferred to a 96-well microtiter plate containing Vero-CCL81 cells. After 48 hours of incubation at 37°C, the extent of GFP fluorescence indicative of viral replication was determined by microscopy. Antibody titers were calculated as the log2 of the reciprocal of the highest serum dilution at which no virus replication was indicated. The obtained VN titers are expressed as mean va...
Embodiment 2
[0032] Example 2 Effect of Adjuvant Type
[0033] Research design
[0034] Twenty piglets seronegative for PEDV were available for this test. Piglets were vaccinated via the intramuscular (IM) route when they were about five weeks old and boosted 2 weeks thereafter. All applications were given into the neck. The vaccine contains 1.5·10 in 2 ml 7 TCID 50 Killed whole PEDV (cell culture-adapted DR13) was inactivated with 1 mM bisethyleneimine (BEI). Blood samples were collected immediately before and one, two, three, four and five weeks after the first vaccination. Sera were collected from blood samples and assayed for virus neutralizing antibody titers. Take rectal temperature on the day of each vaccination and thereafter.
[0035] Animals were divided into four groups of five animals each (see Table 1). The first group received a killed whole virus vaccine formulated in the commercially available adjuvant XSolve™ (available from MSD Animal Health, Boxmeer, The Nethe...
Embodiment 3
[0046] Example 3 Influence of Non-live Antigen Type
[0047] Research design
[0048] Sixty piglets seronegative for PEDV were available for this trial. When piglets were about five weeks old, they were immunized with 2 ml of vaccine via the intramuscular (IM) route and boosted 3 weeks thereafter. All applications were given into the neck. An overview of the experimental groups is given in Table 3. Blood samples were collected before and two, three, four, five and six weeks after the first vaccination. Sera were collected from blood samples and assayed for virus neutralizing antibody titers. Rectal temperature was measured on the day of vaccination and 4 and 24 hours thereafter. After each vaccination, piglets were palpated on the vaccination site at 2, 4, 6, 8, 10, 12 and 14 days post-vaccination.
[0049] table 3. Overview of animal groups and their handling
[0050]
[0051] Materials and methods
[0052] subunit vaccine
[0053] The antigen present in th...
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