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Production and characterization of rabbit polyclonal sera against Shiga toxins Stx1 and Stx2 for detection of Shiga toxin-producing Escherichia coli

Mendes-Ledesma, Meire R. B. ; Rocha, Letícia B. ; Bueris, Vanessa ; Krause, Gladys ; Beutin, Lothar ; Franzolin, Marcia R. ; Trabulsi, Luiz R. ; Elias, Waldir P. ; Piazza, Roxane M. F.

Microbiology and immunology, 2008-10, Vol.52 (10), p.484-491 [Periódico revisado por pares]

Melbourne, Australia: Blackwell Publishing Asia

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Título:
Production and characterization of rabbit polyclonal sera against Shiga toxins Stx1 and Stx2 for detection of Shiga toxin-producing Escherichia coli
Autor: Mendes-Ledesma, Meire R. B. ; Rocha, Letícia B. ; Bueris, Vanessa ; Krause, Gladys ; Beutin, Lothar ; Franzolin, Marcia R. ; Trabulsi, Luiz R. ; Elias, Waldir P. ; Piazza, Roxane M. F.
Assuntos: Animals ; Antibodies, Bacterial - analysis ; Antibodies, Bacterial - biosynthesis ; Cell Survival ; Cercopithecus aethiops ; DNA, Bacterial - chemistry ; DNA, Bacterial - genetics ; ELISA ; Enzyme-Linked Immunosorbent Assay - methods ; Escherichia coli Infections - microbiology ; Humans ; Immunoblotting ; immunodot ; polyclonal rabbit antisera ; Polymerase Chain Reaction ; Rabbits ; Shiga Toxin 1 - analysis ; Shiga Toxin 1 - immunology ; Shiga Toxin 2 - analysis ; Shiga Toxin 2 - immunology ; Shiga toxin-producing E. coli ; Shiga-Toxigenic Escherichia coli - genetics ; Shiga-Toxigenic Escherichia coli - isolation & purification ; Shiga-Toxigenic Escherichia coli - pathogenicity ; Vero Cells ; Virulence
É parte de: Microbiology and immunology, 2008-10, Vol.52 (10), p.484-491
Notas: ArticleID:MIM068
istex:1565DDF0B158633ED085D0365759600EC7D75266
ark:/67375/WNG-6Z2HJCCR-K
In memoriam
Descrição: ABSTRACT STEC has emerged as an important group of enteric pathogens worldwide. In this study, rabbit polyclonal Stx1 and Stx2 antisera were raised and employed in the standardization of immunoassays for STEC detection. Using their respective antisera, the limit of detection of the toxin was 35.0 pg for Stx1 and 5.4 pg for Stx2. By immunoblotting, these antisera recognized both toxin subunits. Cross‐reactivity was observed in the A subunit, but only Stx2 antiserum was able to neutralize the cytotoxicity of both toxins in the Vero cell assay. Six stx‐harboring E. coli isolates were analyzed for their virulence traits. They belonged to different serotypes, including the O48:H7, described for the first time in Brazil. Only three strains harbored eae, and the e‐hly gene and hemolytic activity was detected in five strains. Three isolates showed new stx2 variants (stx2v‐ha and stx2vb‐hb). The ELISA assay detected all six isolates, including one VCA‐negative isolate, while the immunodot assay failed to detect one isolate, which was VCA‐positive. In contrast, the colony‐immunoblot assay detected only one VCA‐positive isolate. Our results demonstrate that among the immunoassays developed in this study, the immunodot, and particularly the ELISA, appear as perspective for STEC detection in developing countries.
Editor: Melbourne, Australia: Blackwell Publishing Asia
Idioma: Inglês

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Production and characterization of rabbit polyclonal sera against Shiga toxins Stx1 and Stx2 for detection of Shiga toxin-producing Escherichia coli

Mendes-Ledesma, Meire R. B. ; Rocha, Letícia B. ; Bueris, Vanessa ; Krause, Gladys ; Beutin, Lothar ; Franzolin, Marcia R. ; Trabulsi, Luiz R. ; Elias, Waldir P. ; Piazza, Roxane M. F.

Melbourne, Australia: Blackwell Publishing Asia

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