Limits...
Development of a blocking ELISA based on a monoclonal antibody against a predominant epitope in non-structural protein 3B2 of foot-and-mouth disease virus for differentiating infected from vaccinated animals.

Fu Y, Lu Z, Li P, Cao Y, Sun P, Tian M, Wang N, Bao H, Bai X, Li D, Chen Y, Liu Z - PLoS ONE (2014)

Bottom Line: The parameters for this SPB-ELISA were established by screening panels of sera of different origins.Serum samples with a percent inhibition (PI) greater than or equal to 46% were considered to be from infected animals, and a PI lower than 46% was considered to indicate a non-infected animal.This is the first description of the conserved and predominant GPYAGPMER epitope of 3B and also the first report of a DIVA test for FMDV NSP 3B based on a McAb against this epitope.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Science, Lanzhou, Gansu, China.

ABSTRACT
A monoclonal antibody (McAb) against non-structural protein (NSP) 3B of foot-mouth-disease virus (FMDV) (3B4B1) was generated and shown to recognize a conserved epitope spanning amino acids 24-32 of 3B (GPYAGPMER) by peptide screening ELISA. This epitope was further shown to be a unique and predominant B cell epitope in 3B2, as sera from animals infected with different serotypes of FMDV blocked the ability of McAb 3B4B1 to bind to NSP 2C3AB. Also, a polyclonal antibody against NSP 2C was produced in a rabbit vaccinated with 2C epitope regions expressed in E. coli. Using McAb 3B4B1 and the 2C polyclonal antibody, a solid-phase blocking ELISA (SPB-ELISA) was developed for the detection of antibodies against NSP 2C3AB to distinguish FMDV-infected from vaccinated animals (DIVA test). The parameters for this SPB-ELISA were established by screening panels of sera of different origins. Serum samples with a percent inhibition (PI) greater than or equal to 46% were considered to be from infected animals, and a PI lower than 46% was considered to indicate a non-infected animal. This test showed a similar performance as the commercially available PrioCHECK NS ELISA. This is the first description of the conserved and predominant GPYAGPMER epitope of 3B and also the first report of a DIVA test for FMDV NSP 3B based on a McAb against this epitope.

Show MeSH

Related in: MedlinePlus

Frequency distribution of PI values obtained by SPB-ELISA.(A) Sera from cattle, (B) sera from sheep, and (C) sera from swine.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4219772&req=5

pone-0111737-g002: Frequency distribution of PI values obtained by SPB-ELISA.(A) Sera from cattle, (B) sera from sheep, and (C) sera from swine.

Mentions: The cutoff PI value for the SPB-ELISA was determined by testing 437 serum samples from healthy unvaccinated animals and 236 serum samples from infected animals. The frequency distributions of the resultant PI values are shown in Figure 2A–C. Table 2 shows the specificity and sensitivity of the assay at three different cutoff values, 40%, 46%, and 50%. Using a cutoff value of 46%, the sensitivity and specificity of the assay were relatively high in the three animal species. Based on these data, serum samples with a PI equal to or greater than 46% were considered to be from infected animals, and samples with a PI lower than 46% were considered to be from non-infected animals. As a quality control measure taken in this assay, the PIs of the positive and weakly positive controls were required to be greater than 70% and 50%, respectively.


Development of a blocking ELISA based on a monoclonal antibody against a predominant epitope in non-structural protein 3B2 of foot-and-mouth disease virus for differentiating infected from vaccinated animals.

Fu Y, Lu Z, Li P, Cao Y, Sun P, Tian M, Wang N, Bao H, Bai X, Li D, Chen Y, Liu Z - PLoS ONE (2014)

Frequency distribution of PI values obtained by SPB-ELISA.(A) Sera from cattle, (B) sera from sheep, and (C) sera from swine.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4219772&req=5

pone-0111737-g002: Frequency distribution of PI values obtained by SPB-ELISA.(A) Sera from cattle, (B) sera from sheep, and (C) sera from swine.
Mentions: The cutoff PI value for the SPB-ELISA was determined by testing 437 serum samples from healthy unvaccinated animals and 236 serum samples from infected animals. The frequency distributions of the resultant PI values are shown in Figure 2A–C. Table 2 shows the specificity and sensitivity of the assay at three different cutoff values, 40%, 46%, and 50%. Using a cutoff value of 46%, the sensitivity and specificity of the assay were relatively high in the three animal species. Based on these data, serum samples with a PI equal to or greater than 46% were considered to be from infected animals, and samples with a PI lower than 46% were considered to be from non-infected animals. As a quality control measure taken in this assay, the PIs of the positive and weakly positive controls were required to be greater than 70% and 50%, respectively.

Bottom Line: The parameters for this SPB-ELISA were established by screening panels of sera of different origins.Serum samples with a percent inhibition (PI) greater than or equal to 46% were considered to be from infected animals, and a PI lower than 46% was considered to indicate a non-infected animal.This is the first description of the conserved and predominant GPYAGPMER epitope of 3B and also the first report of a DIVA test for FMDV NSP 3B based on a McAb against this epitope.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Science, Lanzhou, Gansu, China.

ABSTRACT
A monoclonal antibody (McAb) against non-structural protein (NSP) 3B of foot-mouth-disease virus (FMDV) (3B4B1) was generated and shown to recognize a conserved epitope spanning amino acids 24-32 of 3B (GPYAGPMER) by peptide screening ELISA. This epitope was further shown to be a unique and predominant B cell epitope in 3B2, as sera from animals infected with different serotypes of FMDV blocked the ability of McAb 3B4B1 to bind to NSP 2C3AB. Also, a polyclonal antibody against NSP 2C was produced in a rabbit vaccinated with 2C epitope regions expressed in E. coli. Using McAb 3B4B1 and the 2C polyclonal antibody, a solid-phase blocking ELISA (SPB-ELISA) was developed for the detection of antibodies against NSP 2C3AB to distinguish FMDV-infected from vaccinated animals (DIVA test). The parameters for this SPB-ELISA were established by screening panels of sera of different origins. Serum samples with a percent inhibition (PI) greater than or equal to 46% were considered to be from infected animals, and a PI lower than 46% was considered to indicate a non-infected animal. This test showed a similar performance as the commercially available PrioCHECK NS ELISA. This is the first description of the conserved and predominant GPYAGPMER epitope of 3B and also the first report of a DIVA test for FMDV NSP 3B based on a McAb against this epitope.

Show MeSH
Related in: MedlinePlus