Comparison of a liquid-phase blocking sandwich ELISA and a serum neutralization test to evaluate immunity in potency tests of foot-and-mouth disease vaccines

A liquid-phase blocking sandwich enzyme-linked immunosorbent assay (ELISA-3D) was developed to detect specific antibodies to the 3D protein in sera from foot-and-mouth disease (FMD) virus (FMDV)-infected animals. The assay uses a nonstructural 3D recombinant protein and two polyclonal antisera, one for capture (bovine) and the other for detector (guinea pig). The specificity of the assay was demonstrated by negative results with 101 sera of cattle from the FMD-free zone in Argentina and with bovine and porcine sera raised against various RNA and DNA viruses. The ELISA-3D was able to detect antibodies in cattle after natural or experimental infection with FMDV of A, O, or C types as early as 5 days postinfection and at later stages in persistently infected animals. Comparison of the results with those obtained with the routinely used agar gel immunodiffussion test and a previously described ELISA, both employing a partially purified virus-infection-associated antigen, shows that the ELISA-3D is highly sensitive and specific and gives reproducible results. Its use as a tool for monitoring viral activity and for certification of FMDV-free animals is recommended.

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Comparison of Test Methodologies for Foot-and-Mouth Disease Virus Serotype A Vaccine Matching

Clinical and Vaccine Immunology ◽

10.1128/cvi.00034-14 ◽

2014 ◽

Vol 21 (5) ◽

pp. 674-683 ◽

Cited By ~ 13

Author(s):

Tesfaalem Tekleghiorghis ◽

Klaas Weerdmeester ◽

Froukje van Hemert-Kluitenberg ◽

Rob J. M. Moormann ◽

Aldo Dekker

Keyword(s):

Liquid Phase ◽

Roc Analysis ◽

Neutralization Test ◽

Foot And Mouth Disease ◽

Test Methods ◽

Mouth Disease ◽

Blocking Elisa ◽

Virus Serotype ◽

Discriminatory Capacity

ABSTRACTVaccination has been one of the most important interventions in disease prevention and control. The impact of vaccination largely depends on the quality and suitability of the chosen vaccine. To determine the suitability of a vaccine strain, antigenic matching is usually studied byin vitroanalysis. In this study, we performed threein vitrotest methods to determine which one gives the lowest variability and the highest discriminatory capacity. Binary ethylenimine inactivated vaccines, prepared from 10 different foot-and-mouth disease (FMD) virus serotype A strains, were used to vaccinate cattle (5 animals for each strain). The antibody titers in blood serum samples 3 weeks postvaccination (w.p.v.) were determined by a virus neutralization test, neutralization index test, and liquid-phase blocking enzyme-linked immunosorbent assay (ELISA). The titers were then used to calculate relationship coefficient (r1) values. Theser1values were compared to the genetic lineage using receiver operating characteristic (ROC) analysis. In the two neutralization test methods, the median titers observed against the test strains differed considerably, and the sera of the vaccinated animals did not always show the highest titers against their respective homologous virus strains. When the titers were corrected for test strain effect (scaling), the variability (standard error of the mean per vaccinated group) increased because the results were on a different scale, but the discriminatory capacity improved. An ROC analysis of ther1value calculated on both observed and scaled titers showed that onlyr1values of the liquid-phase blocking ELISA gave a consistent statistically significant result. Under the conditions of the present study, the liquid-phase blocking ELISA showed less variation and still had a higher discriminatory capacity than the other tests.

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Molecular epidemiology and phylogenetic analysis of foot and mouth disease virus type ‘O’ and evaluation of its carrier state in cattle of Assam by Real time PCR

Indian Journal of Animal Research ◽

10.18805/ijar.b-3670 ◽

2018 ◽

Author(s):

Sangeeta Baro ◽

Krishna Sharma ◽

Biswajyoti Sharma ◽

Shantanu Tamuly ◽

P. Deka ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Real Time ◽

Disease Virus ◽

Real Time Pcr ◽

Sandwich Elisa ◽

Foot And Mouth Disease ◽

Carrier State ◽

Mouth Disease ◽

Mouth Disease Virus ◽

Foot And Mouth

The molecular epidemiological study of foot-and-mouth disease virus (FMDV) has been carried out from different outbreaks in Assam the present study is based on the nucleotide sequencingof circulating FMDV serotype. The samples were subjected to sandwich ELISA, multiplex-PCR and molecular phylogeny to identify the type species. The phylogenetic analysis of virus sequence revealed similarity with theBangladesh isolates in the major branching pattern. The serotype ‘O’has found to be dominant and responsible for most of the recentoutbreaks.Thepersistence of serotype ‘O’ and cytokines expression of IL-1á, IL-1â, IFN-á, TNF-á in blood of recovered animals were done by Real time PCR. The findings indicated that IL-1á, IFN-á and TNF-á genes were up-regulated upto 3 months post infection but IL-1â found to be down regulated with progression of recovery. The present study thus supports that real-time PCR is a powerful technique for reliable detection of persistent FMDV in recovered animals.

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foot and mouth disease in some areas of Nainawa province during may 2007

The Iraqi Journal of Veterinary Medicine ◽

10.30539/iraqijvm.v32i2.752 ◽

2008 ◽

Vol 32 (2) ◽

pp. 181-203

Author(s):

Zaid Salah Hussein

Keyword(s):

Liquid Phase ◽

Disease Virus ◽

Foot And Mouth Disease ◽

Mouth Disease ◽

Special Test ◽

Mouth Disease Virus ◽

Foot And Mouth ◽

Immunological Test ◽

Immune Assay

For the determination of FMD infection in Nainawa province Serumsamples were collected from 92 suspected cases (42sheep,50cattle)thiswas performed in September 2007 and included nine region(Talafar,Basheca, Al-shamal, Hamamalaleel ,Cokajlly ,Zamaar ,Al-Namrood,AL-Koosh and Hemedaat), these samples were send frozen to the labs ofstate Vet. Company in Al-Nahdaa , the special test FMD-3ABC bo Ovenzyme immune assay on sheep serums showed that 30.95% werepositive to the test and 19.05% were ambiguous while the test on bovineserums showed increase in both positive and ambiguous results(42%,26% respectively), 8 serums that showed the highest positive resultto the former test were submitted to the special immunological test FMDElisa Kit Liquid Phase Blocking Immunoassay for detection of antibodiesof foot and mouth disease virus test which revealed that all sores of thesamples were infected with O serotype FMD

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