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A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.

A High-throughput Platform for the Screening of Salmonella spp./Shigella spp.

Published on: November 7, 2018

Click Detect: A Rapid and Sensitive Assay for Shiga Toxin 2 Detection.

Benjamin M Thomas¹, Emma L Webb², Katherine L Yan³

¹Genetics and Genomics Interdisciplinary Program, Texas A&M University, College Station, TX 77843, USA.

|December 24, 2025

Summary

This summary is machine-generated.

A new diagnostic platform, Click Detect, rapidly creates sensing probes for detecting Shiga toxin-producing E. coli (STEC) and its toxins. This cost-effective method shows promise for food safety and medical diagnostics.

Keywords:

DARPin Shiga toxin click display diagnostic environmental nanobody

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Area of Science:

Biotechnology
Food Safety
Molecular Diagnostics

Background:

Shiga toxin-producing Escherichia coli (STEC) is a significant foodborne pathogen causing severe gastrointestinal illness and hemolytic uremic syndrome (HUS).
Accurate and rapid detection of STEC and its toxins is crucial for public health and food safety surveillance.

Purpose of the Study:

To develop and validate a novel diagnostic platform, Click Detect, for efficient and sensitive detection of Shiga toxin 2 (Stx2).
To assess the performance of Click Detect in complex matrices relevant to food and environmental monitoring.

Main Methods:

Utilized click display technology for rapid, in vitro generation of uniform protein-cDNA conjugates (sensing probes).
Employed a sandwich-style assay format with click-displayed DARPin (D20) as the sensing probe and nanobody (NG1) as the capture reagent.
Quantified Stx2 using quantitative PCR (qPCR) and loop-mediated isothermal amplification (LAMP).

Main Results:

Click Detect achieved high sensitivity, detecting Stx2 at 600 fM by qPCR and 6 pM by LAMP.
The assay demonstrated robustness, maintaining sensitivity in matrices containing up to 40% swimming pool water or lettuce extract.
Sensing probe preparation was rapid (2 hours), simple, and cost-effective (~USD 0.04 per assay).

Conclusions:

Click Detect offers a rapid, cost-effective, and versatile platform for detecting foodborne pathogens and toxins.
The technology has significant potential for food/environmental monitoring, point-of-care diagnostics, and broader applications in toxin and protein detection.