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Harmonisation of methods for the monitoring of VTEC and subtyping of stx2 genes
Project
Project code: BfR-BIOS-08-1322-443
Contract period: 01.04.2010
- 31.12.2010
Purpose of research: Applied research
The European member States are required in accordance with the Directive 2003/99/EC (EC, 2003) to collect comparable data for harmonise monitoring and reporting of verotoxigenic E. coli (VTEC) in animals and foodstuffs. For this reason and on the basis of the opinion of the panel of Biological Hazards for the monitoring of VTEC (on behalf of the EFSA), uniform technical specifications have been proposed for the monitoring and reporting of VTEC. The data collected using these methods would facilitate a better analysis of the situation at Member State levels.The AFSA has proposed guidelines for the detection of VTEC O157 in animals and food-stuffs because food is one of the most important sources of VTEC infections and VTEC O157 is the serogroup most often reported in human VTEC infections. These guidelines can also be used for the detection of other VTEC serogroups as: O26, O113, O111 and O145The detection of E: coli O157 is based on the ISO 16654:2001 method, the isolated O157 strains must be verified by PCR for the presence of Shigatoxin genes (stx1 and stx2) and for the intimin gene (eae). For the detection of other VTEC serogroups the use of the draft CEN TC275/WG6, currently submitted to ISO for evaluation, is proposed. In this draft the detection of VTEC non-O157 is based on conventional PCR as well as real-time PCR methods, followed by a confirmation step for the isolation of the STEC strains. Different PCR methods for detection and characterization of VTEC have been developed and tested in the NRL E. coli at the BfR. The PCR methods developed in the NRL E. coli should be compared with the methods descriibed in the draft CEN TC275/WG6 so that we can see which methods are the most appropriate to recognize all variants of VTEC and whether the procedures described in the ISO with the CEN Methods can be accepted by the NRL E. coli for reasons of harmonization. Additionally, PCR primers for the detection of Shiga toxin variants 1, 2, and 2c, and 2dactivatable
Section overview
Subjects
- Animal health
- Food microbiology
- Toxicology