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