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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.
Biosensors
|December 24, 2025
Summary
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.
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.
