AbstractsBiology & Animal Science

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

by Johanna Tawe




Institution: Uppsala University
Department:
Year: 2014
Keywords: Pathogenic E. coli; STEC; VTEC; EHEC; PCR; validation; Engineering and Technology; Teknik och teknologier; Civilingenjörsprogrammet i molekylär bioteknik; Molecular Biotechnology Engineering Programme
Record ID: 1330860
Full text PDF: http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-220478


Abstract

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.