Simple Diagnostic Test for STEC

October 10, 2014

Laboratory Services Division, University of Guelph
MMD
Senior Technician

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Company:

SA Scientific

Product Name:

Verotoxin-producing Escherichia coli Detection Kit

Catalog Number:

E-6210

Our lab is in the pathogen detection field, we tested the Verotoxin-producing Escherichia coli Detection Kit from SA Scientific to test for the presence of shiga toxin producing E. coli (STEC) in food samples. The kit was chosen for its ease of setup and quick detection. The Kit uses the Verotoxin (VT, also known as Shiga toxin) genes as the target sequences (stx1 and stx2). The kit comes with all reagents needed for DNA extraction and setting the reaction with controls. I also purchased the Fluorescence Detection Reagent for quick qualitative identification. The reactions all worked as predicted, as examined by either fluorescence or agarose gel analysis.

Experimental Design and Results Summary

Application

Shiga Toxin producing E. coli detection

Starting Material

Spiked Food Samples

Protocol Overview

Food Samples were spiked with STEC and incubated overnight. The extraction Fluid from the kit was used for DNA extraction and the LAMP reaction was set up using the reagents supplied. A positive control was used that was included in the kit. Analysis was conducted by visualizing fluorescence in tubes and agarose gel electrophoresis.

Tips

Run an agarose gel for the first few experiments to ensure you are obtaining amplification.

Results Summary

The results obtained were as expected, with amplification in the control reaction and no amplification in either a no template control or a no spike control. Fluorescence was easily detected qualitatively in the tube when using the Fluorescence Detection Reagent or via agarose gel electrophoresis.

Additional Notes

The Fluorescence Detection Reagent allows for instant detection and the ability to skip electrophoresis.

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Summary

The Good

Quick, easy to set up, no false positives or negatives.

The Bad

A little on the expensive side.

The Bottom Line

The bottom line is this product works exactly as advertised and is easy to set up. Anyone with good sterile technique and novice experience with molecular biology could use this kit.

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