Student Research

Real‐time, Isothermal Detection of Shiga Toxin‐Producing E. coli Using Recombinase Polymerase Amplification

This study is significant to the detection of foodborne pathogens which affect the food industry in the US and world‐wide. Shiga toxin (Stx) producing E. coli (STEC) were detected using Recombinase Polymerase Amplification (RPA). The detection was done isothermally (39ºC), and in real‐time, using specifically designed primers and probes, which amplified and detected the Shiga toxin1‐(Stx 1) and Stx 2‐encoding genes that encodethe major STEC virulence factors. The primers and probes were used to identify presence or absence of stx1 and stx2 in STEC (n = 12), non‐STEC E. coli (n = 28), and other bacterial genera (n = 7). This study was the first to successfully detect STEC using RPA, and in real‐time (within 10 min), and had a sensitivity of 5 to 50 CFU/mL. RPA is a quick, and portable technique, which has potential for direct use, e.g., in produce fields or at the point‐of‐care. Future studies will focus on evaluating RPA on produce spiked with known concentrations of STEC, and later on produce in the field and environmental samples.

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