Next Article in Journal
Fatal Neurotoxicosis in Dogs Associated with Tychoplanktic, Anatoxin-a Producing Tychonema sp. in Mesotrophic Lake Tegel, Berlin
Previous Article in Journal
Contractile Response of Bovine Lateral Saphenous Vein to Ergotamine Tartrate Exposed to Different Concentrations of Molecularly Imprinted Polymer
Previous Article in Special Issue
Implementing the Bruker MALDI Biotyper in the Public Health Laboratory for C. botulinum Neurotoxin Detection
Article Menu
Issue 2 (February) cover image

Export Article

Open AccessFeature PaperArticle
Toxins 2018, 10(2), 59; https://doi.org/10.3390/toxins10020059

An Improved Method for the Sensitive Detection of Shiga Toxin 2 in Human Serum

1
Western Regional Research Center, U.S. Department of Agriculture, Agricultural Research Service, 800 Buchanan Street, Albany, CA 94710, USA
2
Center for HUS Control, Prevention and Management, Fondazione IRCCS Ca’Granda Ospedale Maggiore Policlinico, Via Commenda 9, 20122 Milano, Italy
3
Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Via San Giacomo 14, 40126 Bologna, Italy
*
Authors to whom correspondence should be addressed.
Received: 18 November 2017 / Revised: 12 January 2018 / Accepted: 29 January 2018 / Published: 31 January 2018
(This article belongs to the Collection Rapid Detection of Bacterial Toxins)
View Full-Text   |   Download PDF [1880 KB, uploaded 26 February 2018]   |  

Abstract

Shiga toxins (Stx) released by Stx-producing E. coli (STEC) are virulence factors that are most closely associated with hemolytic uremic syndrome (HUS), a life-threatening complication of intestinal infections by STEC. Stx have to enter into the circulatory system before they are delivered to target organs and cause damage. The presence of Stx in sera could be a risk indicator for HUS development. However, the detection of Stx, particularly Stx2, has been difficult due to the presence of Stx2-binding components in human serum. Here, we report new ELISA-based methods for the detection of Stx1 and Stx2 in human serum and the effect of guanidinium chloride on enhancing the sensitivity for the detection of Stx2. The recovery rate for Stx2 was 62% when Stx2-spiked serum samples were treated with guanidinium chloride at a concentration of 200 mM, in contrast to 17% without guanidinium chloride treatment. The effectiveness of guanidinium chloride treatment for the detection of Stx2 in human serum was validated using sera from STEC-infected patients. Coimmunoprecipitation results indicated a specific physical interaction between Stx2 and the human serum amyloid P component (HuSAP) in human serum samples. Our in vitro study demonstrated that the inhibition from HuSAP alone for the detection of Stx2 was only 20%, much less than 69.6% from human serum at Stx2 level 10 ng/mL, suggesting that there may be other factors that bind Stx2 in human serum. This study indicates that treatment of serum samples with guanidinium chloride may be useful for the early and sensitive detection of Stx2 in sera of STEC-infected patients, so preventive measures can be adopted in a timely manner. View Full-Text
Keywords: co-immunoprecipitation; enzyme-linked immunosorbent assay; guanidinium chloride; hemolytic uremic syndrome; human serum amyloid P component; Shiga toxins co-immunoprecipitation; enzyme-linked immunosorbent assay; guanidinium chloride; hemolytic uremic syndrome; human serum amyloid P component; Shiga toxins
Figures

Figure 1

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).

Share & Cite This Article

MDPI and ACS Style

He, X.; Ardissino, G.; Patfield, S.; Cheng, L.W.; Silva, C.J.; Brigotti, M. An Improved Method for the Sensitive Detection of Shiga Toxin 2 in Human Serum. Toxins 2018, 10, 59.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Toxins EISSN 2072-6651 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top