An ELISA based on the repeated foot-and-mouth disease virus 3B epitope peptide can distinguish infected and vaccinated cattle

Gao, Mingchun; Zhang, Runxiang; Li, Meng; Li, Shuang; Cao, Yanghui; Ma, Bo; Wang, Junwei

doi:10.1007/s00253-011-3815-0

Cited by 31 publications

(22 citation statements)

References 26 publications

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“…Tests for the detection of FMDV-positive antisera have been developed using synthetic 3B peptides; however, this is at a cost premium and is limited by the length of peptides that can be produced. A lower-cost alternative is the use of recombinant 3B peptides produced in E. coli (17,40,46,48). By extension, the use of tandem repeats of 3B peptides to effectively multiply the number of potential antibody binding sites on the antigen has been used successfully to differentiate infected and vaccinated animals (18,48).…”

Section: Discussionmentioning

confidence: 99%

“…A lower-cost alternative is the use of recombinant 3B peptides produced in E. coli (17,40,46,48). By extension, the use of tandem repeats of 3B peptides to effectively multiply the number of potential antibody binding sites on the antigen has been used successfully to differentiate infected and vaccinated animals (18,48). In addition, others have developed DIVA tests by expressing the entire 3B protein individually or as part of a larger FMDV NSP protein, typically 3AB, 2C3AB, or 3ABC (for recent examples, see references 31,45,[49][50][51].…”

Section: Discussionmentioning

confidence: 99%

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Pan-Serotype Diagnostic for Foot-and-Mouth Disease Using the Consensus Antigen of Nonstructural Protein 3B

et al. 2015

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bAn amino acid consensus sequence for the seven serotypes of foot-and-mouth disease virus (FMDV) nonstructural protein 3B, including all three contiguous repeats, and its use in the development of a pan-serotype diagnostic test for all seven FMDV serotypes are described. The amino acid consensus sequence of the 3B protein was determined from a multiple-sequence alignment of 125 sequences of 3B. The consensus 3B (c3B) protein was expressed as a soluble recombinant fusion protein with maltosebinding protein (MBP) using a bacterial expression system and was affinity purified using amylose resin. The MBP-c3B protein was used as the antigen in the development of a competition enzyme-linked immunosorbent assay (cELISA) for detection of anti-3B antibodies in bovine sera. The comparative diagnostic sensitivity and specificity at 47% inhibition were estimated to be 87.22% and 93.15%, respectively. Reactivity of c3B with bovine sera representing the seven FMDV serotypes demonstrated the pan-serotype diagnostic capability of this bioreagent. The consensus antigen and competition ELISA are described here as candidates for a pan-serotype diagnostic test for FMDV infection. Foot-and-mouth disease (FMD) is the most contagious viral vesicular disease that affects cloven-hoofed livestock species. FMD has significant global socioeconomic consequences, from national livestock industries suffering because of international trade restrictions to subsistence farmers suffering because of losses of stock productivity and livelihoods. Western European, North American, and Far East Asian/Pacific regions and most South American countries have official recognition of freedom-from-FMD status, with or without vaccination, by the World Organization for Animal Health (OIE). Regions in which FMD remains endemic tend to be those of lesser economic capacity, and this limits their ability to control or to eradicate the disease (1, 2). As a result, FMD remains an ongoing problem in regions in which it is endemic, and it is a persistent threat to regions that are free of the disease. Seven serotypes have been described for foot-and-mouth disease virus (FMDV), i.e., O, A, C, Asia 1, and South African Territories 1 (SAT1), SAT2, and SAT3. The disease forms produced by each serotype are clinically indistinguishable (3); similarly, FMD is clinically indistinguishable from other vesicular lesion-causing diseases, such as vesicular stomatitis, swine vesicular exanthema, and swine vesicular disease, and laboratory-based clinical diagnosis is required (4, 5). FMD infections may be mild or subclinical in ovine or caprine species, further complicating clinical diagnosis (6, 7). Exposure to one serotype does not confer cross-serotype immunity, potentially complicating diagnosis when multiple serotypes are circulating during an outbreak (8). A definitive diagnosis of FMD is possible only with laboratory testing.Conventional serological assays for FMD, including the virus neutralization test, liquid-phase blocking enzyme-linked immunosorbent assay (ELISA), and so...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Pan-Serotype Diagnostic for Foot-and-Mouth Disease Using the Consensus Antigen of Nonstructural Protein 3B

et al. 2015

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“…The FMDV NSP 3B exists as three similar but non-identical tandem repeats, 3B1 (VPg1), 3B2 (VPg2), and 3B3 (VPg3), that share a high amino acid sequence homology and a common core amino acid motif [QKPL (M) K] [3]. These proteins were found to have a high density of linear B cell epitopes, some of which, including GPYAGPMERQKPLK, PMERQKPLKVKAKA, and QKPLKVKAKAPVVK, are conserved and immunodominant and could potentially be used to develop DIVA tests [8].…”

Section: Introductionmentioning

confidence: 99%

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

Lu²,

Li³

et al. 2014

PLoS ONE

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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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“…The reaction was stopped after 15 min by adding 1 M H 2 SO 4 and the OD450 nm of each well was read using a microplate reader (Molecular Devices, Sunnyvale, CA). (12) Basic characterization of MAb…”

Section: Cell Fusionmentioning

confidence: 99%

A Novel Monoclonal Antibody Against the Constant Region of Goose Immunoglobulin Light Chain

Guo

Gao²,

Ma³

et al. 2014

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

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An ELISA based on the repeated foot-and-mouth disease virus 3B epitope peptide can distinguish infected and vaccinated cattle

Cited by 31 publications

References 26 publications

Pan-Serotype Diagnostic for Foot-and-Mouth Disease Using the Consensus Antigen of Nonstructural Protein 3B

Pan-Serotype Diagnostic for Foot-and-Mouth Disease Using the Consensus Antigen of Nonstructural Protein 3B

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

A Novel Monoclonal Antibody Against the Constant Region of Goose Immunoglobulin Light Chain

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