The main food-borne pathogens of concern in the beef chain are Shiga toxin-producing Escherichia coli (STEC) and Salmonella; other pathogens, including Listeria monocytogenes and Campylobacter spp. may also be present. Furthermore, the occurrence and development of antimicrobial-resistant (AMR) pathogens in the beef chain is a public health issue. Traditional culture-based methods are laborious and time consuming. Rapid methods, including nucleic acid-, immunologic- and biosensor-based techniques can be very sensitive and specific and provide more timely information regarding the presence of pathogens in the beef chain. This chapter reviews these methods, as well as recent advances in next-generation sequencing technologies.