Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates
Enterotoxigenic Escherichia coli (ETEC) produce heat-labile (LT) and/or heat-stable enterotoxins (ST). Despite that, the mechanism of action of both toxins are well known, there is great controversy in the literature concerning the in vitro production and release of LT and, for ST, no major concern...
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doaj-b3fbdad3604d4e3d8bb7e32d04f82cce2020-11-24T20:44:46ZengMDPI AGToxins2072-66512013-12-015122384240210.3390/toxins5122384toxins5122384Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli IsolatesLetícia B. Rocha0Christiane Y. Ozaki1Denise S. P. Q. Horton2Caroline A. Menezes3Anderson Silva4Irene Fernandes5Fabio C. Magnoli6Tania M. I. Vaz7Beatriz E. C. Guth8Roxane M. F. Piazza9Bacteriology Laboratory, Butantan Institute, São Paulo, SP 05503-900, BrazilBacteriology Laboratory, Butantan Institute, São Paulo, SP 05503-900, BrazilBacteriology Laboratory, Butantan Institute, São Paulo, SP 05503-900, BrazilBacteriology Laboratory, Butantan Institute, São Paulo, SP 05503-900, BrazilBacteriology Laboratory, Butantan Institute, São Paulo, SP 05503-900, BrazilImmunopathology Laboratory, Butantan Institute, São Paulo, SP 05503-900, BrazilImmunochemistry Laboratory, Butantan Institute, São Paulo, SP 05503-900, BrazilBacteriology Section, Adolfo Lutz Institute, São Paulo, SP 01246-000, BrazilDepartment of Microbiology, Immunology, Parasitology, Escola Paulista de Medicina, Federal University of São Paulo, SP 04923-062, BrazilBacteriology Laboratory, Butantan Institute, São Paulo, SP 05503-900, BrazilEnterotoxigenic Escherichia coli (ETEC) produce heat-labile (LT) and/or heat-stable enterotoxins (ST). Despite that, the mechanism of action of both toxins are well known, there is great controversy in the literature concerning the in vitro production and release of LT and, for ST, no major concerns have been discussed. Furthermore, the majority of published papers describe the use of only one or a few ETEC isolates to define the production and release of these toxins, which hinders the detection of ETEC by phenotypic approaches. Thus, the present study was undertaken to obtain a better understanding of ST and LT toxin production and release under laboratory conditions. Accordingly, a collection of 90 LT-, ST-, and ST/LT-producing ETEC isolates was used to determine a protocol for toxin production and release aimed at ETEC detection. For this, we used previously raised anti-LT antibodies and the anti-ST monoclonal and polyclonal antibodies described herein. The presence of bile salts and the use of certain antibiotics improved ETEC toxin production/release. Triton X-100, as chemical treatment, proved to be an alternative method for toxin release. Consequently, a common protocol that can increase the production and release of LT and ST toxins could facilitate and enhance the sensitivity of diagnostic tests for ETEC using the raised and described antibodies in the present work.http://www.mdpi.com/2072-6651/5/12/2384ETECheat-labile toxinheat-stable toxinproductionreleasedetection |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Letícia B. Rocha Christiane Y. Ozaki Denise S. P. Q. Horton Caroline A. Menezes Anderson Silva Irene Fernandes Fabio C. Magnoli Tania M. I. Vaz Beatriz E. C. Guth Roxane M. F. Piazza |
spellingShingle |
Letícia B. Rocha Christiane Y. Ozaki Denise S. P. Q. Horton Caroline A. Menezes Anderson Silva Irene Fernandes Fabio C. Magnoli Tania M. I. Vaz Beatriz E. C. Guth Roxane M. F. Piazza Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates Toxins ETEC heat-labile toxin heat-stable toxin production release detection |
author_facet |
Letícia B. Rocha Christiane Y. Ozaki Denise S. P. Q. Horton Caroline A. Menezes Anderson Silva Irene Fernandes Fabio C. Magnoli Tania M. I. Vaz Beatriz E. C. Guth Roxane M. F. Piazza |
author_sort |
Letícia B. Rocha |
title |
Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates |
title_short |
Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates |
title_full |
Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates |
title_fullStr |
Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates |
title_full_unstemmed |
Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates |
title_sort |
different assay conditions for detecting the production and release of heat-labile and heat-stable toxins in enterotoxigenic escherichia coli isolates |
publisher |
MDPI AG |
series |
Toxins |
issn |
2072-6651 |
publishDate |
2013-12-01 |
description |
Enterotoxigenic Escherichia coli (ETEC) produce heat-labile (LT) and/or heat-stable enterotoxins (ST). Despite that, the mechanism of action of both toxins are well known, there is great controversy in the literature concerning the in vitro production and release of LT and, for ST, no major concerns have been discussed. Furthermore, the majority of published papers describe the use of only one or a few ETEC isolates to define the production and release of these toxins, which hinders the detection of ETEC by phenotypic approaches. Thus, the present study was undertaken to obtain a better understanding of ST and LT toxin production and release under laboratory conditions. Accordingly, a collection of 90 LT-, ST-, and ST/LT-producing ETEC isolates was used to determine a protocol for toxin production and release aimed at ETEC detection. For this, we used previously raised anti-LT antibodies and the anti-ST monoclonal and polyclonal antibodies described herein. The presence of bile salts and the use of certain antibiotics improved ETEC toxin production/release. Triton X-100, as chemical treatment, proved to be an alternative method for toxin release. Consequently, a common protocol that can increase the production and release of LT and ST toxins could facilitate and enhance the sensitivity of diagnostic tests for ETEC using the raised and described antibodies in the present work. |
topic |
ETEC heat-labile toxin heat-stable toxin production release detection |
url |
http://www.mdpi.com/2072-6651/5/12/2384 |
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