26 - 30 April 2010, Zagreb, Croatia

Course Material

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Lectures

Monday, 26 April 2010

• Molecular typing – overview. Alex Friedrich
• Improving the understanding of the micro and macro epidemiology of bacterial pathogens using genetic typing approaches (part 1). Hajo Grundmann
• Improving the understanding of the micro and macro epidemiology of bacterial pathogens using genetic typing approaches (part 2). Hajo Grundmann.
• PFGE – principles and applications. Ana Budimir
• PCR-based typing methods. Raquel Sa-Leao

Tuesday, 27 April 2010

• Multipe-Locus Variable number tandem repeats Analysis; A tool to study bacterial populations structures and perform molecular epidemiology. Leo Schouls
• Type-Ned, a pilot to develop a structured molecular typing network in the Netherlands. Leo Schouls
• Molecular epidemiology of E. coli.  Branka Bedenic

Wednesday, 28 April 2010

• Significance of typing in clinical laboratory and practical use in infection control. Alex Friedrich
• Molecular epidemiology of Streptococcus pneumoniae. Raquel Sa-Leao
• Typing networks: Practical added value for regional and cross-border molecular surveillance. Alex Friedrich
• Introduction to spa and MLST. Dimitar Nashev

Thursday, 29 April 2010

• SCCmec typing  in MRSA and CoNS. Frederic Laurent
• Data analysis in PFGE.  Koen Janssens
• Quantitative Comparison of Molecular Typing Techniques. Dag Harmsen

Friday, 30 April 2010

• Molecular typing of Staphylococcus aureus using DNA microarray. Frederic Laurent
• Getting and analyzing DNA sequences. Dag Harmsen
• MLST & SLST: Technical Background. Dag Harmsen
• Outlook: From Surveillance Data to Knowledge - Early Warning Systems & GIS. Dag Harmsen
• ESBLs. Branka Bedenic

Practical Sessions

No presentations available for practicals.

• DNA preparation for PCR
• DNA preparation for PFGE (cell lysis and proteinase K digestion in agarose plugs)
• Preparation of PCR reaction mixtures for spa typing, SCCmec, MLVA and MLST
• Agarose gel electrophoresis of PCR products
• Staining and photographing of PCR agarose gels
• Purification of PCR products
• Preparation of sequencing reaction mixtures for spa typing and MLST
• Restriction endonuclease digestion of DNA for PFGE
• PFGE agarose gel preparation, sample loading and gel electrophoresis
• Analysis of spa and MLST sequencing results (trace files)
• Analysis of MLVA PCR results (gel images and automated DNA sequencer data)
• Analysis of SCCmec PCR results
• Staining and photographing PFGE agarose gels
• Analysis of PFGE results (gel images)
• Participants’ presentations - describing and discussing experimental results

Course Faculty

• Ana Budimir, Zagreb, Croatia
• Branka Bedenic, Zagreb, Croatia
• Alexander Friedrich, Münster, Germany
• Hajo Grundmann, Bilthoven, The Netherlands
• Sara Hæggman, Solna, Sweden
• Dag Harmsen, Münster, Germany
• Smilja Kalenic, Zagreb, Croatia
• Frederic Laurent, Lyon, France
• Dimitar Nashev, Sofia, Bulgaria
• Raquel Sá-Leão, Oeiras, Portugal
• Leo Schouls, Bilthoven, The Netherlands

Dubravko Sijak
Clinical Hospital Centre Zagreb
Department of Clinical and Molecular Microbiology
Kispaticeva 12
10 000 Zagreb, Croatia
Phone +385 2367 318
Dubravko Sijak

Course Objectives

To provide participants with theoretical and practical information - including hands-on experience - on epidemiological typing methods. The practical part of the course focuses on PFGE and PCR-based typing methods (e.g., spa, SCCmec, MLVA and MLST) of Staphylococcus aureus, ESBL-producing Enterobacteriaceae, non-fermentative Gram-negatives (Pseudomonas and Acinetobacter), enterococci, and Streptococcus pneumoniae.

Course Venue

University of Zagreb, School of Medicine
Clinical Hospital Centre Zagreb
Dept of Clinical and Molecular Microbiology
Kispaticeva 12
10 000 Zagreb, Croatia
Phone +385 2367 318

Target Audience

16 - 22 participants comprising trainees in clinical microbiology and infectious diseases and molecular biologists working in clinical microbiology.