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Toxins 2015, 7(11), 4564-4576; doi:10.3390/toxins7114564

A Rapid and Sensitive Method to Measure the Functional Activity of Shiga Toxins in Human Serum

1
Dipartimento di Medicina Specialistica, Diagnostica e Sperimentale, Sede di Patologia Generale, Università di Bologna, Via San Giacomo 14, 40126 Bologna, Italy
2
Center for HUS Control, Prevention and Management, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, 20122 Milan, Italy
3
Unit of Microbiology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, 20122 Milan, Italy
4
Istituto Superiore di Sanità, 00161 Rome, Italy
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Xiaohua He
Received: 21 September 2015 / Revised: 26 October 2015 / Accepted: 26 October 2015 / Published: 4 November 2015
(This article belongs to the Collection Rapid Detection of Bacterial Toxins)
View Full-Text   |   Download PDF [841 KB, uploaded 4 November 2015]   |  

Abstract

Shiga toxins (Stx) have a definite role in the development of hemolytic uremic syndrome in children with hemorrhagic colitis caused by pathogenic Stx-producing Escherichia coli (STEC) strains. The dramatic effects of these toxins on the microvasculature of different organs, particularly of the kidney, are well known, whereas there is no consensus on the mechanism by which Stx reach the endothelia of target organs and/or indirectly injure these body sites. We hereby describe a quick (4 h), radioactive, Raji cell-based method designed for the detection of Stx in human sera. The assay monitors the translation impairment induced by these powerful inhibitors of protein synthesis, which are identified properly by neutralizing their activity with specific monoclonal antibodies. By this method, we detected for the first time the functional activity of Stx in sera of STEC-infected patients during hemorrhagic colitis. Recent research has pointed to a dynamic process of Stx-induced renal intoxication in which concurrent and interactive steps are involved. Our rapid and specific method could be useful for studying the kinetics of Stx during the natural course of STEC infection and the interplay between Stx activity in serum and Stx presence in different blood fractions (neutrophils, monocytes, platelets, leukocyte-platelet aggregates, microvesicles, lipoproteins). View Full-Text
Keywords: hemolytic uremic syndrome; Shiga toxin-producing Escherichia coli; eukaryotic protein synthesis; Raji cells hemolytic uremic syndrome; Shiga toxin-producing Escherichia coli; eukaryotic protein synthesis; Raji cells
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Arfilli, V.; Carnicelli, D.; Ardissino, G.; Torresani, E.; Scavia, G.; Brigotti, M. A Rapid and Sensitive Method to Measure the Functional Activity of Shiga Toxins in Human Serum. Toxins 2015, 7, 4564-4576.

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