Validation of PCR assays for detection of Shiga toxin-producing E. coli O104:H4 and O121 in food

Shigatoxin-producing Escherichia coli (STEC) can cause infections in humans which can beserious and sometimes fatal. There is a great need for methods that are able to detect differentserogroups of STEC. In this project, conventional and real-time PCR assays for detection ofSTEC O104:H4 and O121, as recommended by the European Union Reference Laboratory(EU-RL) for STEC, were validated. The specificity, limit of detection, repeatability,efficiency and robustness were determined for three real-time PCR assays. The validationshowed that the real-time PCR reactions were specific and sensitive although some additionaltests are required