Update on the STEC Network preparedness for molecular typing data collection (PT-PFGE5)
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1 12th Annual workshop of the EU Reference Laboratories for E. coli, Rome October 2017 Update on the STEC Network preparedness for molecular typing data collection (PT-PFGE5) Antonella Maugliani EU Reference Laboratory for Escherichia coli including VTEC Dept. of Food Safety, Nutrition and Veterinary Public Health Istituto Superiore di Sanità, Rome, Italy
2 SETTING-UP PFGE SOPs produced by EURLs for STEC, Listeria monocytogenes and Salmonella describes all the details for the production, interpretation and curation of the PFGE profiles nts/704e.pdf ; /702e.pdf ; / 703e.pdf Technical Specification on molecular typing data collection, on metadata and on the ownership of the data PRESENT EFSA DATA-BASE is a reality Several MS nominated their rapresentative for submitting molecular typing data (11MS for 2016) The curators are ready too
3 The EURL-VTEC activities for the preparedness on molecular typing Proficiency tests (EQA) on PFGE (5 rounds in ) THE PT-PFGE6 STRAINS WILL BE SENT NEXT MONTH Training sessions on PFGE ( ) 20 NRLs TRAINED Basic courses on BioNumerics software ( )
4 THE AIMS EQA ON PFGE: useful tool to asses the capability to produce the good quality profiles The database for non human PFGE profiles should contain high quality profiles
5 The EURL-VTEC Proficiency testing activity on PFGE 1. PT-PFGE1 (with PT10, January-March 2013) 16 EU NRLs + Norway, Switzerland and Egypt 11 E.coli strains Conducted jointly with the ECDC-FWD network 2. PT-PFGE2 (with PT11, June-September 2013) 23 EU NRLs + Norway and Switzerland 6 E.coli strains 3. PT-PFGE3 (with PT13, April-June 2014) 25 EU NRLs + Norway and Switzerland 7 E.coli strains 4. PT-PFGE4 (April-June 2015) 24 EU NRLs + Norway, Switzerland, Macedonia and Russia 10 E. coli strains Conducted jointly with the ECDC-FWD network 5. PT-PFGE5 (November-December 2016) 17 EU NRLs + Norway and Turkey 10 E. coli strains, in the framework of the PT18
6 EQA on molecular typing: evaluation NRL s Proficiency PT-PFGE1 and PT-PFGE2: PulseNet Procedure PNQ01(visual assessment) PT-PFGE3, PT-PFGE4 AND PT-PFGE5 : EFSA SOP for PFGE profile interpretation (BioNumerics assisted) PT-PFGE6: EFSA SOP Rating: P F G E o o r a i r o o d x c e l l e n t
7 PT-PFGE5 RESULTS - 1 SUBMITTED DATA TIFF: GEL IMAGE FILE + EXCEL FILE XML: FILE GENERATED BY THE SOFTWARE BIONUMERICS AFTER THE ANALISYS PERFORMED BY THE NRL
8 PT-PFGE5 RESULTS - 2 EXCELLENT FAIR POOR REGARDLESS THE FORMAT FILE
9 CRITICAL POINTS PT-PFGE5
10 BAND ASSIGNMENT SAMPLE IDENTIFICATION ALWAYS CHECK THE POSITION OF THE STRAINS WITHIN THE LANES ALWAYS VERIFY THE CORRECT IDENTIFICATION OF THE STRAIN WITHIN EACH LANE IT S CRUCIAL FOR THE IDENTIFICATION OF THE STRAIN PROFILE...
11 PFGE Proficiency testing (EQA): Trend from 2012 NRLs performance in producing PFGE profiles
12 The EURL-VTEC training activities for molecular typing 10 Training sessions on PFGE ( ) (in total 24 colleagues: 20 NRLs + Norway + Algeria + 2 non NRLs ) 3 Courses on BioNumerics software use participants + 2 observers from Nigeria and USA joint with the three EURLs (E. coli-salmonella Listeria) 12 participants from the three pathogens vet-networks joint with the three EURLs 12 participants from the three pathogens vet-networks the fourth edition will be organized by EURL-Salmonella joint with all the three EURLs in the Summer 2018
13 Collaboration between the three EURLs (STEC, Listeria monocytogenes and Salmonella) has been established The main objective is the HARMONIZATION COMMON BN COURSE ( )
14 Many labs continue to apply for the training on PFGE - Importance of the participation in the PT after the training session (55% of the participating labs improved their performance) Proficiency test activity is a very useful instrument either for the design of the SOPs used for the evaluation either for the assessment of the preparedness of the network for the EFSA-DB 79% of the participants in the PT-PFGE5 accomplishing an excellent result Collaboration between vet-eurls has been established to consolidate the network on BN analysis (two JOINT- BN courses have performed and one more is planned) The identification of the critical points will be used by the EURL-VTEC to find the areas needing dedicated training sessions
15 THANK YOU
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