2003
DOI: 10.1590/s1517-83822003000500002
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Polyclonal anti-intimin antibody: immunological characterization and its use in EPEC and EHEC diagnosis

Abstract: Intimins are outer membrane proteins expressed by enteric bacterial pathogens capable of inducing intestinal attachment-and-effacement lesion (A/E). Through immunoblotting, immunofluorescence, flow citometry and immunogold we observed that the obtained polyclonal antibody against conserved intimin region recognizes the different intimin subtypes and suggests that it can be used as a tool for EPEC and EHEC detection. Besides, immuno-dot assay seems to be a possible alternative as a capture method.

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Cited by 13 publications
(16 citation statements)
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“…The sections were deparaffinized and incubated with 0.3% H 2 O 2 in PBS for 20 min at room temperature to inactivate endogenous peroxidase. Samples were blocked with PBS containing 2% bovine serum albumin (BSA) for 30 min at room temperature and incubated for 2 h in the same buffer containing a 1:5,000 dilution of polyclonal rabbit anti-intimin IgG-enriched fraction (41,46). Sections were washed three times with PBS and incubated for 1 h at room temperature with biotin-labeled goat anti-rabbit IgG (Sigma) diluted in PBS-2% BSA.…”
Section: Methodsmentioning
confidence: 99%
“…The sections were deparaffinized and incubated with 0.3% H 2 O 2 in PBS for 20 min at room temperature to inactivate endogenous peroxidase. Samples were blocked with PBS containing 2% bovine serum albumin (BSA) for 30 min at room temperature and incubated for 2 h in the same buffer containing a 1:5,000 dilution of polyclonal rabbit anti-intimin IgG-enriched fraction (41,46). Sections were washed three times with PBS and incubated for 1 h at room temperature with biotin-labeled goat anti-rabbit IgG (Sigma) diluted in PBS-2% BSA.…”
Section: Methodsmentioning
confidence: 99%
“…Concerning intimin detection, a rabbit polyclonal sera raised against the conserved region of intimin (Int388-667) 143 was employed in order to detect tEPEC isolates expressing α, β, γ, δ and ɛ intimin reported an application of immunoblotting with 100% specificity and 97% sensitivity in the detection of eae positive E. coli strains 144, 145, 146. These authors clearly demonstrated that polyclonal rabbit antisera is suitable for immunoblotting as a diagnostic tool, and showed that protein denaturation and linearization is a critical step for anti-intimin antibody accessibility.…”
Section: Typical and Atypical Enteropathogenic E Colimentioning
confidence: 99%
“…In a previous study, the reactivity of IgG‐enriched fraction of rabbit anti‐intimin serum with tEPEC belonging to serotypes that express α, β, γ, δ and ε intimin by immunoblotting had been observed. This reactivity was confirmed using immuno‐dot assay, immunofluorescence, flow cytometry and immunogold labelling (Koga et al. 2003).…”
Section: Discussionmentioning
confidence: 78%
“…Amongst the methods for the detection of virulence factor expression, the immunoassays can be considered as first alternative, and for these purposes polyclonal and/or Mabs are raised (Koga et al. 2003; Menezes et al.…”
Section: Discussionmentioning
confidence: 99%