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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.

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Detection of antibodies by SPB-ELISA and two commercial ELISA kits (3ABC-ELISA and PrioCHECK FMDV-NS ELISA) in sera collected sequentially from animals infected with FMDV.(A)–(D) show the results from cattle no. 4009, 0738, 4017, and 4004, respectively, (E) and (F) show the results from sheep no. 54 and 40, and (G) shows the result from pig no. 39.
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pone-0111737-g003: Detection of antibodies by SPB-ELISA and two commercial ELISA kits (3ABC-ELISA and PrioCHECK FMDV-NS ELISA) in sera collected sequentially from animals infected with FMDV.(A)–(D) show the results from cattle no. 4009, 0738, 4017, and 4004, respectively, (E) and (F) show the results from sheep no. 54 and 40, and (G) shows the result from pig no. 39.

Mentions: Results from 97 sequential serum samples from four cattle (n = 40), two sheep (n = 41), and one swine (n = 16) infected with FMDV are shown in Figure 3A–G. Of the 40 serum samples from four infected cattle, 39 samples yielded identical results by the SPB-ELISA, 3ABC-ELISA, and PrioCHECK NS ELISA. Only one sample, taken from animal no. 4004 at 8 DPI, showed a positive result by SPB-ELISA (Fig. 3D). Of the 41 serum samples from two infected sheep, only one differential result, taken from animal no. 54 at 164 DPI, was observed between the SPB-ELISA and the PrioCHECK NS ELISA (Fig. 3E); however, differential results were observed between the 3ABC-ELISA and the other two ELISAs in four samples: the sample taken from sheep no. 40 at 164 DPI (Fig. 3F), and the samples taken from sheep no. 54 at 117, 132, and 194 DPI (Fig. 3E). Of the 16 sera from an infected swine, one negative result at 7 DPI by the SPB-ELISA, one negative result at 181 DPI by the PrioCHECK NS ELISA, and three negative results, at 22, 50, and 181 DPI, by the 3ABC-ELISA were observed (Fig. 3G).


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)

Detection of antibodies by SPB-ELISA and two commercial ELISA kits (3ABC-ELISA and PrioCHECK FMDV-NS ELISA) in sera collected sequentially from animals infected with FMDV.(A)–(D) show the results from cattle no. 4009, 0738, 4017, and 4004, respectively, (E) and (F) show the results from sheep no. 54 and 40, and (G) shows the result from pig no. 39.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0111737-g003: Detection of antibodies by SPB-ELISA and two commercial ELISA kits (3ABC-ELISA and PrioCHECK FMDV-NS ELISA) in sera collected sequentially from animals infected with FMDV.(A)–(D) show the results from cattle no. 4009, 0738, 4017, and 4004, respectively, (E) and (F) show the results from sheep no. 54 and 40, and (G) shows the result from pig no. 39.
Mentions: Results from 97 sequential serum samples from four cattle (n = 40), two sheep (n = 41), and one swine (n = 16) infected with FMDV are shown in Figure 3A–G. Of the 40 serum samples from four infected cattle, 39 samples yielded identical results by the SPB-ELISA, 3ABC-ELISA, and PrioCHECK NS ELISA. Only one sample, taken from animal no. 4004 at 8 DPI, showed a positive result by SPB-ELISA (Fig. 3D). Of the 41 serum samples from two infected sheep, only one differential result, taken from animal no. 54 at 164 DPI, was observed between the SPB-ELISA and the PrioCHECK NS ELISA (Fig. 3E); however, differential results were observed between the 3ABC-ELISA and the other two ELISAs in four samples: the sample taken from sheep no. 40 at 164 DPI (Fig. 3F), and the samples taken from sheep no. 54 at 117, 132, and 194 DPI (Fig. 3E). Of the 16 sera from an infected swine, one negative result at 7 DPI by the SPB-ELISA, one negative result at 181 DPI by the PrioCHECK NS ELISA, and three negative results, at 22, 50, and 181 DPI, by the 3ABC-ELISA were observed (Fig. 3G).

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