New immuno-PCR assay for detection of low concentrations of shiga toxin 2 and its variants.

New immuno-PCR assay for detection of low concentrations of shiga toxin 2 and its variants. Research Abstract Details 

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New immuno-PCR assay for detection of low concentrations of shiga toxin 2 and its variants. Abstract Text:

Wenlan Zhang,Martina Bielaszewska,Matthias Pulz,Karsten Becker,Alexander W Friedrich,Helge Karch,Thorsten Kuczius,Wenlan Zhang,Martina Bielaszewska,Matthias Pulz,Karsten Becker,Alexander W Friedrich,Helge Karch,Thorsten Kuczius,

Shiga toxin (Stx)-producing Escherichia coli (STEC) strains secrete toxins that are major virulence factors and diagnostic targets, but some STEC strains secrete Stx in amounts that cannot be detected using conventional cell cytotoxicity or immunological assays. Therefore, there is an urgent need for more-sensitive Stx detection methods. We describe the development of an assay that can detect low concentrations of Stx2 and its variants. An immuno-PCR Stx2 assay was developed based on an enzyme immunoassay (EIA) combining antibody capture and DNA amplification to increase the signal. The immuno-PCR assay detected 10 pg/ml of purified Stx2, compared to 1 ng/ml Stx2 detected by commercial EIA. Consequently, immuno-PCR detected Stx2 and its variants in STEC strains that produce the toxins at levels that are nondetectable by using the EIA, as well as the Stx2 in EIA-negative enriched stool cultures from patients. Our data demonstrate that the immuno-PCR developed here is a highly sensitive and specific method for the detection of trace amounts of Stx2 and Stx2 variants. It is therefore suitable for use by clinical microbiological laboratories to improve the toxin detection in clinical samples.

New immuno-PCR assay for detection of low concentrations of shiga toxin 2 and its variants. Publishing Authors By Initials

W Zhang,M Bielaszewska,M Pulz,K Becker,AW Friedrich,H Karch,T Kuczius,W Zhang,M Bielaszewska,M Pulz,K Becker,AW Friedrich,H Karch,T Kuczius,

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New immuno-PCR assay for detection of low concentrations of shiga toxin 2 and its variants. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of clinical microbiology

VOLUME: 46

Page Numbers: 1292-7

Journal Abbreviation: J. Clin. Microbiol.

ISSN: 1098-660X

DAY: 13

MONTH: 02

YEAR: 2008

New immuno-PCR assay for detection of low concentrations of shiga toxin 2 and its variants. Information

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LANGUAGE: eng

NlmUniqueID: 7505564

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AFFILIATION: Institute for Hygiene, University of Münster, Robert Koch Str. 41, 48149 Münster, Germany. tkuczius@uni-muenster.de.

Country: United States

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MEDLINETA: J Clin Microbiol

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