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Performance of ELISAs for detection of antibodies against porcine respiratory and reproductive syndrome virus in serum of pigs after PRRSV type 2 live vaccination and challenge.

ABSTRACT: BACKGROUND:The aim of the study was to evaluate the performance of different newly developed and/or commercially available ELISAs for detection of PRRSV specific antibodies. Consequently, ten PRRSV negative piglets (group V) were vaccinated with a PRRSV type 2 vaccine. Blood samples were taken before as well as seven, 21 and 42 days after vaccination. At day 42 after vaccination (day 0 of the study) all of the piglets from group V and 10 non-prevaccinated PRRSV negative piglets (group N) were challenged with an HP PRRSV type 2 field strain. Blood samples were taken before and at days 3, 7, 10, 14, 21 and 28 after challenge. The success of vaccination and challenge was measured with RT qPCR. All serum samples were tested with six ELISAs for detection of PRRSV antibodies. Three of them are nucleocapsid-based, two use a glycoprotein extract and one uses inactivated whole virus as antigen. The specificity of the ELISAs was evaluated using 301 serum samples of piglets from PRRSV negative herds. RESULTS:The piglets from group V tested positive by RT qPCR at day 7 after vaccination and all piglets tested positive at day 3 after challenge. PRRSV specific antibodies were seen with all nucleocapsid-based ELISAs from day 21 after vaccination onwards in group V and from day 10 after challenge in group N. The glycoprotein-based ELISAs detected antibodies from day 42 after vaccination (group V) and day 21 after challenge (group N). The agreement according to kappa-coefficient was almost perfect. The glycoprotein-based ELISAs were able to distinguish PRRSV type 2, although with some cross reactions. Regarding specificity, the ELISAs performed differently (specificity between 97.4 % and 100 %), whereas most of the ELISAs with higher sensitivity had a slightly lower specificity. CONCLUSIONS:All tested ELISA were able to detect PRRSV antibodies in the serum of pigs vaccinated with a PRRSV type 2 vaccine and after challenge with an HP PRRSV type 2 field strain. The onset on antibody detection differed, depending on the type of antigen used in the ELISAs. Most of the ELISAs with a higher sensitivity had a lower specificity.

SUBMITTER: Sattler T

PROVIDER: S-EPMC5382508 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Performance of ELISAs for detection of antibodies against porcine respiratory and reproductive syndrome virus in serum of pigs after PRRSV type 2 live vaccination and challenge.

Sattler Tatjana T Pikalo Jutta J Wodak Eveline E Schmoll Friedrich F

Porcine health management 20151211

<h4>Background</h4>The aim of the study was to evaluate the performance of different newly developed and/or commercially available ELISAs for detection of PRRSV specific antibodies. Consequently, ten PRRSV negative piglets (group V) were vaccinated with a PRRSV type 2 vaccine. Blood samples were taken before as well as seven, 21 and 42 days after vaccination. At day 42 after vaccination (day 0 of the study) all of the piglets from group V and 10 non-prevaccinated PRRSV negative piglets (group N) ...[more]

PMID: 28405425

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Project description:BackgroundIn this study, six enzyme-linked immunosorbent assays (ELISA), intended for routine porcine reproductive and respiratory syndrome virus (PRRSV) herd monitoring, are tested for their ability to detect PRRSV specific antibodies in the serum of pigs after vaccination with an inactivated PRRSV type 1 vaccine and subsequent infection with a highly pathogenic (HP) PRRSV field strain. For this reason, ten piglets (group V) from a PRRSV negative herd were vaccinated twice at the age of 2 and 4 weeks with an inactivated PRRSV vaccine. Ten additional piglets (group N) from the same herd remained unvaccinated. Three weeks after second vaccination, each of the piglets received an intradermal application of an HP PRRSV field strain. Serum samples were taken before first vaccination as well as before and 3, 7, 10 and 14 days after HP PRRSV application. All serum samples were tested for PRRSV RNA by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) as well as for PRRSV antibodies with all six study ELISAs.ResultsAt the beginning of the study (before vaccination), all of the piglets were PRRSV antibody negative with all study ELISAs. They also tested negative for PRRSV RNA measured by RT-qPCR. From day 3 after HP PRRSV application until the end of the study, a viremia was detected by RT-qPCR in all of the piglets. On day 0 (day of HP PRRSV application), nine out of ten piglets of the pre-vaccinated group tested PRRSV antibody positive with one of the tested ELISAs, although with lower S/P values than after infection. On day 10 after HP PRRSV application, all study ELISAs except one had significantly higher S/P or OD values, respectively more positive samples, in group V than in group N.ConclusionsOnly one of the tested ELISAs was able to detect reliably PRRSV antibodies in pigs vaccinated with an inactivated PRRSV vaccine. With most of the tested ELISAs, higher S/P values respectively more positive samples after PRRSV infection were seen in the pre-vaccinated group than in the non-vaccinated.

Project description:Cell-mediated immunity (CMI), IL-10, and the protective efficacy of modified-live porcine reproductive and respiratory syndrome virus (PRRSV) vaccines (MLV) against co-challenge with PRRSV-1 and PRRSV-2 (HP-PRRSV) were investigated. Seventy, PRRSV-free, 3-week old, pigs were allocated into 7 groups. Six groups were intramuscularly vaccinated with MLV, including Porcilis (PRRSV-1 MLV, MSD Animal Health, The Netherlands), Amervac (PRRSV-1 MLV, Laboratorios Hipra, Spain), Fostera (PRRSV-2 MLV, Zoetis, USA), Ingelvac PRRS MLV and Ingelvac PRRS ATP (PRRSV-2, Boehringer Ingelheim, USA), and Prime Pac PRRS (PRRSV-2 MLV, MSD Animal Health, The Netherlands). Unvaccinated pigs were left as control. Lymphocyte proliferative response, IL-10 and IFN-γ production were determined. At 35 days post-vaccination (DPV), all pigs were inoculated intranasally with 2 ml of each PRRSV-1 (105.4 TCID50/ml) and PRRSV-2 (105.2 TCID50/ml, HP-PRRSV). Following challenge, sera were quantitatively assayed for PRRSV RNA. Pigs were necropsied at 7 days post-challenge. Viremia, macro- and microscopic lung lesion together with PRRSV antigen presence were evaluated in lung tissues. The results demonstrated that, regardless of vaccine genotype, CMI induced by all MLVs was relatively slow. Increased production of IL-10 in all vaccinated groups was observed at 7 and 14 DPV. Pigs in Amervac, Ingelvac MLV and Ingelvac ATP groups had significantly higher levels of IL-10 compared to Porcilis, Fostera and Prime Pac groups at 7 and 14 DPV. Following challenge, regardless to vaccine genotype, vaccinated pigs had significantly lower lung lesion scores and PRRSV antigens than those in the control group. Both PRRSV-1 and PRRSV-2 RNA were significantly reduced. Prime Pac pigs had lowest PRRSV-1 and PRRSV-2 RNA in serum, and micro- and macroscopic lung lesion scores (p < 0.05) compared to other vaccinated groups. In conclusion, PRRSV MLVs, regardless of vaccine genotype, can reduce viremia and lung lesions following co-challenge with PRRSV-1 and PRRSV-2 (HP-PRRSV). The main difference between PRRSV MLV is the production of IL-10 following vaccination.

Dataset Information

Performance of ELISAs for detection of antibodies against porcine respiratory and reproductive syndrome virus in serum of pigs after PRRSV type 2 live vaccination and challenge.

Publications

Performance of ELISAs for detection of antibodies against porcine respiratory and reproductive syndrome virus in serum of pigs after PRRSV type 2 live vaccination and challenge.

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