Molecular Biotechnology Programme
Uppsala University School of Engineering
UPTEC X 11 035 Date of issue 2011-06
Author
Mattias Carlsson
Title (English)
Optimization and validation of realtime-PCR assay for detection of verocytotoxing-producing E. coli (VTEC)
Title (Swedish)
Optimering och validering av realtids-PCR assay för detektion av verocytotoxin-producerande E. coli (VTEC)
Abstract
Pathogenic E. coli such as VTEC can cause serious diseases to humans. A detection method for VTEC based on realtime-PCR has been developed by EU-RL VTEC (European Union Reference Laboratory VTEC) and the National Food Administration. In this master thesis I have optimized and validated a part of the method which detects serotypes of VTEC. During optimization the primer concentration, probe concentration and annealing temperature were evaluated for a more sensitive detection. Validation showed that the PCR-reaction is both effective and sensitive (LOD100). Validation of the whole method did not give any significant results due to the fact the sensitivity (LOD50) could not be determined.
Keywords
VTEC (STEC, EHEC), serotypes, realtime-PCR, optimization, validation Supervisors
Catarina Nilsson
National Food Administration
Scientific reviewer
Lars-Göran Josefsson Uppsala universitet
Project name Sponsors
Language
Swedish
Security
ISSN 1401-2138 Classification
Supplementary bibliographical information Pages
40
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