Journal: Porcine Health Management

Article Title: Evaluation of classical swine fever E2 (CSF-E2) subunit vaccine efficacy in the prevention of virus transmission and impact of maternal derived antibody interference in field farm applications

doi: 10.1186/s40813-020-00188-6

Figure Lengend Snippet: Pig groups and experiment schedules in the study

Article Snippet: The dynamics of the CSF-specific antibody level were monitored by using an IDEXX CSFV Ab blocking ELISA test kit and the gray dotted line indicated adequate antibody level (blocking percentage > 40%).

Techniques: Vaccines, Sampling

Journal: Porcine Health Management

Article Title: Evaluation of classical swine fever E2 (CSF-E2) subunit vaccine efficacy in the prevention of virus transmission and impact of maternal derived antibody interference in field farm applications

doi: 10.1186/s40813-020-00188-6

Figure Lengend Snippet: The CSF-E2 subunit vaccine could prevent CSFV horizontal transmission on a vaccination–challenge animal trial. SPF pigs were immunized with CSF-E2 subunit vaccine (group A, circle and black line) or placebo (group C, square and light gray dashed line) and challenged with CSFV ALD strain (1 × 10 5 TCID 50 ) after two vaccinations. Groups A and C were raised in two isolated ABSL-2 units. The sentinel SPF pigs (group B, triangle and gray line) were cohabited with group A at 4 days after the virus challenge to monitor the transmission of CSFV. Pigs in group C were sacrificed 7 days after the virus challenge due to severe clinical symptoms. a The CSFV-specific neutralizing antibody level was analyzed after the CSFV challenge. The gray dotted line indicated the protective neutralizing antibody level (1:32). b The viremia level was monitored after the virus challenge to identify the infection of CSFV. c and d The number of leukocytes and platelets were counted after virus challenge to monitor CSFV infection-induced leukopenia (< 11,000 cells/μL, gray dotted line) and thrombocytopenia (< 211,000 cells/μL, gray dotted line)

Article Snippet: The dynamics of the CSF-specific antibody level were monitored by using an IDEXX CSFV Ab blocking ELISA test kit and the gray dotted line indicated adequate antibody level (blocking percentage > 40%).

Techniques: Transmission Assay, Isolation, Virus, Infection

Journal: Porcine Health Management

Article Title: Evaluation of classical swine fever E2 (CSF-E2) subunit vaccine efficacy in the prevention of virus transmission and impact of maternal derived antibody interference in field farm applications

doi: 10.1186/s40813-020-00188-6

Figure Lengend Snippet: Evaluation of vaccine induced immune response on sows and the passive immunity of piglets. Sows were immunized with two doses of CSF-E2 vaccine at 3 and 5 weeks before parturition (group D, n = 25) or one dose of LPC vaccine before insemination (group E, n = 35). a The CSF-specific antibody level was detected using an IDEXX CSFV Ab test kit to monitor vaccine induced immune response. The gray dotted line indicates a positive antibody response (blocking percentage > 40%). b The saliva CSFV RNA was assessed for 1 month after vaccination using real-time PCR. Pearson’s chi-square test with Yate’s continuity correction showed a significant difference between the CSFV positive count of groups D and E ( p < 0.001). c The piglet serum samples randomly selected from CSF-E2 vaccinated sows (group F, n = 20) and LPC vaccinated sows (group G, n = 20) were analyzed by using the IDEXX CSFV Ab test kit. The mean blocking percentage of each group was calculated and Welch’s two-sample t-test was performed to evaluate the statistical difference between groups. *** p < 0.001

Article Snippet: The dynamics of the CSF-specific antibody level were monitored by using an IDEXX CSFV Ab blocking ELISA test kit and the gray dotted line indicated adequate antibody level (blocking percentage > 40%).

Techniques: Blocking Assay, Real-time Polymerase Chain Reaction

Journal: Porcine Health Management

Article Title: Evaluation of classical swine fever E2 (CSF-E2) subunit vaccine efficacy in the prevention of virus transmission and impact of maternal derived antibody interference in field farm applications

doi: 10.1186/s40813-020-00188-6

Figure Lengend Snippet: The interference of maternally derived antibodies on CSFV vaccines. Piglets with high maternal derived antibody were immunized with two doses of CSF-E2 vaccine (group H, n = 6), LPC vaccine (group I, n = 6), or placebo (group J, n = 6) at 3 and 6 weeks of age and received a booster immunization with one dose of LPC vaccine at 16 weeks of age (*). The dynamics of the CSF-specific antibody level were monitored by using an IDEXX CSFV Ab blocking ELISA test kit and the gray dotted line indicated adequate antibody level (blocking percentage > 40%). Different letters indicate statistically significant difference ( p < 0.001)

Article Snippet: The dynamics of the CSF-specific antibody level were monitored by using an IDEXX CSFV Ab blocking ELISA test kit and the gray dotted line indicated adequate antibody level (blocking percentage > 40%).

Techniques: Derivative Assay, Vaccines, Blocking Assay, Enzyme-linked Immunosorbent Assay

Journal: Porcine Health Management

Article Title: Evaluation of classical swine fever E2 (CSF-E2) subunit vaccine efficacy in the prevention of virus transmission and impact of maternal derived antibody interference in field farm applications

doi: 10.1186/s40813-020-00188-6

Figure Lengend Snippet: Application of CSFV vaccines in a field farm with long-term observation. Pigs were divided into three groups and immunized with one dose of CSF-E2 vaccine (group K, n = 10) or placebo (group M, n = 10) at 4 weeks of age or two doses of LPC vaccine (group L, n = 10) at 12 and 16 weeks of age. The serum samples were collected from weaning to finishing in four-week intervals and analyzed using an IDEXX CSFV Ab blocking ELISA test kit. The gray dotted line represents the adequate antibody level (blocking percentage > 40%) and different letters indicate a statistically significant difference ( p < 0.05)

Article Snippet: The dynamics of the CSF-specific antibody level were monitored by using an IDEXX CSFV Ab blocking ELISA test kit and the gray dotted line indicated adequate antibody level (blocking percentage > 40%).

Techniques: Vaccines, Blocking Assay, Enzyme-linked Immunosorbent Assay

Journal: Virology Journal

Article Title: Generation and immunogenicity analysis of recombinant classical swine fever virus glycoprotein E2 and E rns expressed in baculovirus expression system

doi: 10.1186/s12985-021-01507-1

Figure Lengend Snippet: Expression and characterization of recombinant CSFV E2 and E rns protein. a SDS-PAGE analysis of E2 protein. Lane 1, purified E2 protein; Lane 2, E2 protein treated with PNGase F. b SDS-PAGE analysis of E rns protein. Lane 1, purified E rns protein; Lane 2, E rns protein treated with PNGase F. c Western blot of purified E2 protein. Anti-His antibody was used for the western blot. Lane 1, purified E2 protein; Lane 2, E2 protein treated with PNGase F. d Western blot of purified E rns protein. Anti-His antibody was used for the western blot. Lane 1, purified E rns protein; Lane 2, E rns protein treated with PNGase F

Article Snippet: The presence of E2-specific antibody was determined by blocking ELISA with IDEXX CSFV antibody test kit (IDEXX GmbH, Switzerland) according to the manufacturer’s instructions.

Techniques: Expressing, Recombinant, SDS Page, Purification, Western Blot

Journal: Virology Journal

Article Title: Generation and immunogenicity analysis of recombinant classical swine fever virus glycoprotein E2 and E rns expressed in baculovirus expression system

doi: 10.1186/s12985-021-01507-1

Figure Lengend Snippet: Humoral immune responses in immunized rabbits. Twenty 14-week-old New Zealand white rabbits (four per group) were intramuscularly immunized at days 0 and 28. Serum samples were collected at each week after the first immunization. a The presence of E2-specific antibodies was tested using the IDEXX CSFV antibody test kit. The results were reported as the antibody blocking rate and a rate ≥ 40% was interpreted as positive. b E rns -specific antibodies were detected by ELISA as we described in Materials and Methods. Relative antibody concentrations were determined using the ELISA plate reader at 450 nm. c The titers of CSFV specific neutralization antibodies were determined via immunoperoxidase monolayer assay (IPMA) and the results were expressed as the reciprocal of the highest dilution that caused 50% neutralization

Article Snippet: The presence of E2-specific antibody was determined by blocking ELISA with IDEXX CSFV antibody test kit (IDEXX GmbH, Switzerland) according to the manufacturer’s instructions.

Techniques: Blocking Assay, Enzyme-linked Immunosorbent Assay, Neutralization

Journal: Virology Journal

Article Title: Generation and immunogenicity analysis of recombinant classical swine fever virus glycoprotein E2 and E rns expressed in baculovirus expression system

doi: 10.1186/s12985-021-01507-1

Figure Lengend Snippet: Agarose gel electrophoresis of samples detected by RT-nestPCR assay. The spleens of the rabbits were collected and total RNA was extracted. The viral RNA was detected by the RT-nestPCR amplification. Lane M: DL 100 plus DNA Marker. Lane 1–4: CSFV-E2 group; Lane 5–8: CSFV-E rns group; Lane 9–12: CSFV E2 + E rns group; Lane 13–16: C-strain group; Lane 17–20: PBS group

Article Snippet: The presence of E2-specific antibody was determined by blocking ELISA with IDEXX CSFV antibody test kit (IDEXX GmbH, Switzerland) according to the manufacturer’s instructions.

Techniques: Agarose Gel Electrophoresis, Amplification, Marker

Journal: Toxins

Article Title: Gastric Ulceration and Immune Suppression in Weaned Piglets Associated with Feed-Borne Bacillus cereus and Aspergillus fumigatus

doi: 10.3390/toxins12110703

Figure Lengend Snippet: Detecting Foot and mouth disease (FMD)-specific antibodies post inoculation with B. cereus , A. fumigatus or the combination of B. cereus and A. fumigatus . As for FMD-specific antibodies, a statistically significant decline was found in the B. cereus group compared to the control group on day 7 ( p < 0.05). However, no statistical difference was found among three treated groups on day 14. The data were expressed as the mean ± SD ( n = 9; n = 6). *: p < 0.05; **: p < 0.01.

Article Snippet: CSFV-specific antibodies, PEDV-specific antibodies and FMD-specific antibodies were measured using the specific commercial kit (IDEXX, Beijing, China) according to the manufacturer’s protocol.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: A Genetically Engineered Bivalent Vaccine Coexpressing a Molecular Adjuvant against Classical Swine Fever and Porcine Epidemic Diarrhea

doi: 10.3390/ijms241511954

Figure Lengend Snippet: Humoral immune response and PEDV-specific IFN-γ production induced by recombinant CSFVs in rabbits. Rabbits were inoculated twice intravenously with the indicated virus at a 2-week interval (n = 3). DMEM was used as a negative control (n = 3). The serum of the rabbits was collected at 14 days postbooster, and virus-specific antibodies and neutralizing antibodies were determined by ELISA and neutralizing assay, respectively. PEDV-specific IFN-γ in peripheral blood lymphocytes collected 14 days postbooster was measured using a commercially available cytokine ELISA kit (Mlbio, Shanghai, China). ( A ) CSFV-specific antibody titers. ( B ) CSFV-neutralizing antibody titers. ( C ) PEDV-specific IgG concentration. ( D ) PEDV-neutralizing antibody titers. ( E ) PEDV-specific IFN-γ concentration. The comparisons were performed using Student’s t test, one-way ANOVA or the nonparametric Kruskal–Wallis test. ns, not significant; ** p < 0.01. *** p < 0.001.

Article Snippet: To quantify CSFV-specific antibodies, a commercial ELISA kit from IDEXX (USA) was employed, as mentioned previously [ ].

Techniques: Recombinant, Virus, Negative Control, Enzyme-linked Immunosorbent Assay, Neutralizing Assay, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: A Genetically Engineered Bivalent Vaccine Coexpressing a Molecular Adjuvant against Classical Swine Fever and Porcine Epidemic Diarrhea

doi: 10.3390/ijms241511954

Figure Lengend Snippet: Humoral immune response and PEDV-specific IFN-γ production induced by recombinant CSFVs in pigs. Specific CSFV- and PEDV-free piglets aged 5 weeks were intramuscularly vaccinated twice with the indicated inoculum at a 2-week interval. Sera of the piglets were collected at the indicated timepoints, and virus-specific antibodies and neutralizing antibodies were determined by ELISA and the neutralizing assay. PEDV-specific IFN-γ in PBMCs collected 14 days postbooster was measured. ( A ) CSFV-specific antibody titers. ( B ) CSFV neutralizing antibody titers. ( C ) PEDV-specific IgG concentration. ( D ) PEDV neutralizing antibody titers. ( E ) PEDV-specific IgA concentration. ( F ) PEDV-specific IFN-γ concentration. The comparisons of the values were performed using Student’s t test. ** p < 0.01.

Article Snippet: To quantify CSFV-specific antibodies, a commercial ELISA kit from IDEXX (USA) was employed, as mentioned previously [ ].

Techniques: Recombinant, Virus, Enzyme-linked Immunosorbent Assay, Neutralizing Assay, Concentration Assay

Journal: Veterinary Research

Article Title: Surface displaying of swine IgG1 Fc enhances baculovirus-vectored vaccine efficacy by facilitating viral complement escape and mammalian cell transduction

doi: 10.1186/s13567-017-0434-5

Figure Lengend Snippet: Detection of antibody and cellular immune responses induced by recombinant baculoviruses. A Antibody levels in sera of immunized pigs by indirect ELISA. Four groups of pigs were intramuscularly injected twice with commercial vaccine C-strain, BV-VSVG-ED-pFc-CMV-S/P-E2 or BV-VSVG-ED-CMV-E2 at a 1-week interval. Serum samples from each group were collected weekly and tested. A dashed line indicates the positive cutoff value of ELISA (OD630 nm = 0.35). B IFN-γ levels in sera of immunized pigs by ELISA. Serum samples from each group were collected and tested using a commercial ELISA kit (4A Biotech, Beijing, China) for quantitative detection of pig IFN-γ. Bars with different letters were significantly different ( P < 0.05).

Article Snippet: The CSFV-specific antibodies were tested in sera collected at 0, 7, 14, 21 and 28 days post-infection (dpi) using commercial ELISA kit (Keqian, Wuhan, China), and IFN-γ was assayed in sera collected at 0, 14 and 28 dpi using a commercial ELISA kit (4A Biotech, Beijing, China).

Techniques: Recombinant, Indirect ELISA, Injection, Enzyme-linked Immunosorbent Assay