New Stx2e Monoclonal Antibodies for Immunological Detection and Distinction of Stx2 Subtypes.

Stx2e is a primary virulence factor in STEC strains that cause edema disease in neonatal piglets. Though Stx2a and Stx2e are similar, many antibody-based Stx detection kits are designed to detect Stx2a and do not recognize the Stx2e subtype.Four monoclonal antibodies against Stx2e were developed and...

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Main Authors: Craig Skinner, Stephanie Patfield, Bradley J Hernlem, Xiaohua He
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2015-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC4507848?pdf=render
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author Craig Skinner
Stephanie Patfield
Bradley J Hernlem
Xiaohua He
author_facet Craig Skinner
Stephanie Patfield
Bradley J Hernlem
Xiaohua He
author_sort Craig Skinner
collection DOAJ
description Stx2e is a primary virulence factor in STEC strains that cause edema disease in neonatal piglets. Though Stx2a and Stx2e are similar, many antibody-based Stx detection kits are designed to detect Stx2a and do not recognize the Stx2e subtype.Four monoclonal antibodies against Stx2e were developed and characterized. Two of these mAbs recognize the B subunit of Stx2e, Stx2f, and to a lesser extent, Stx2b, Stx2c, and Stx2d. The other two mAbs recognize the A subunit of Stx2e, and cross-react with all Stx2 subtypes except Stx2f. The most sensitive sandwich ELISA using these mAbs has a limit of detection for Stx2e of 11.8 pg/mL. The ability of the neutralizing antibody Stx2e-2 to block Stx2e-receptor binding in Vero cells was visualized using immunofluorescence. Combinations of these and previously developed mAbs permit ELISA-based differentiation between closely related Stx2a, Stx2c, and Stx2d (using mAbs Stx2-5/2-1, Stx2-5/2e-2, and Stx2e-3/2e-2, respectively).The sensitive immunoassays developed in this study should augment our capacity to detect Stx2e in porcine environments and biological samples. Moreover, immunoassays that can distinguish between the closely related Stx2a, Stx2c, and Stx2d subtypes can be useful in quickly analyzing Stx subtypes in samples containing more than one strain of STEC.
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spelling doaj.art-2e83a58c83144c44862c6997aaf523052022-12-21T23:33:21ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01107e013241910.1371/journal.pone.0132419New Stx2e Monoclonal Antibodies for Immunological Detection and Distinction of Stx2 Subtypes.Craig SkinnerStephanie PatfieldBradley J HernlemXiaohua HeStx2e is a primary virulence factor in STEC strains that cause edema disease in neonatal piglets. Though Stx2a and Stx2e are similar, many antibody-based Stx detection kits are designed to detect Stx2a and do not recognize the Stx2e subtype.Four monoclonal antibodies against Stx2e were developed and characterized. Two of these mAbs recognize the B subunit of Stx2e, Stx2f, and to a lesser extent, Stx2b, Stx2c, and Stx2d. The other two mAbs recognize the A subunit of Stx2e, and cross-react with all Stx2 subtypes except Stx2f. The most sensitive sandwich ELISA using these mAbs has a limit of detection for Stx2e of 11.8 pg/mL. The ability of the neutralizing antibody Stx2e-2 to block Stx2e-receptor binding in Vero cells was visualized using immunofluorescence. Combinations of these and previously developed mAbs permit ELISA-based differentiation between closely related Stx2a, Stx2c, and Stx2d (using mAbs Stx2-5/2-1, Stx2-5/2e-2, and Stx2e-3/2e-2, respectively).The sensitive immunoassays developed in this study should augment our capacity to detect Stx2e in porcine environments and biological samples. Moreover, immunoassays that can distinguish between the closely related Stx2a, Stx2c, and Stx2d subtypes can be useful in quickly analyzing Stx subtypes in samples containing more than one strain of STEC.http://europepmc.org/articles/PMC4507848?pdf=render
spellingShingle Craig Skinner
Stephanie Patfield
Bradley J Hernlem
Xiaohua He
New Stx2e Monoclonal Antibodies for Immunological Detection and Distinction of Stx2 Subtypes.
PLoS ONE
title New Stx2e Monoclonal Antibodies for Immunological Detection and Distinction of Stx2 Subtypes.
title_full New Stx2e Monoclonal Antibodies for Immunological Detection and Distinction of Stx2 Subtypes.
title_fullStr New Stx2e Monoclonal Antibodies for Immunological Detection and Distinction of Stx2 Subtypes.
title_full_unstemmed New Stx2e Monoclonal Antibodies for Immunological Detection and Distinction of Stx2 Subtypes.
title_short New Stx2e Monoclonal Antibodies for Immunological Detection and Distinction of Stx2 Subtypes.
title_sort new stx2e monoclonal antibodies for immunological detection and distinction of stx2 subtypes
url http://europepmc.org/articles/PMC4507848?pdf=render
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