Shiga toxin-producing Escherichia coli (STEC) are pathogenic microorganisms that can significantly impact public health. Consumption of improperly cooked ground meats, raw milk, and raw vegetables have been associated with outbreaks involving STEC. Methods to evaluate their presence in foods are routinely used in food testing laboratories worldwide to assess their prevalence during the food supply chain and thus prevent the occurrence of consumption of contaminated foods. While applications of traditional microbiological methods and molecular methods involving polymerase chain reaction are available, alternative technologies may become valuable diagnostic tools. Loop-mediated isothermal amplification is a specific and rapid method that enables fast amplification of a target sequence at a constant temperature. In this chapter we have included a revision of the different targets to identify relevant STEC serogroups using LAMP in combination with different reporting technologies to visualize LAMP products. In specific, we describe an optimized LAMP method for identification of STEC in various food matrices targeting Shiga toxin genes utilizing bioluminescence real-time detection to visualize the LAMP products.