5th Workshop.
Professor V.J. Benedí
Mechanisms of Antimicrobial Resistance.
A Practical
Approach
Daily Program
Day 1. Sunday, June 20
Arrival and transfer to the
hotel. Transfer to be arranged by each participant.
5th
Workshop Professor V. J. Benedí by Sebastián Albertí.
18:30-19:30 h. Lecture.
Epidemiology of multiresistant nosocomial
bacteria by Marc Struelens.
Day 2. Monday, June
21
08,30-09-30h
Lecture.
Carbapenem resistance by Giussepe Cornaglia.
09.45-10.15 h.
Overview of the laboratory sessions:
TOPIC 1: Antimicrobial
susceptibility testing in P. aeruginosa.
TOPIC 2: DNA
typing of P. aeruginosa isolates.
TOPIC 3: Hypermutation and antimicrobial resistance in P. aeruginosa.
TOPIC 4: Resistance to quinolones in P. aeruginosa.
TOPIC 5: The contribution of b-lactamases to antimicrobial
resistance.
TOPIC 6: The contribution of outer membrane to antimicrobial resistance.
10.15-10.30 h: Objectives of
the day.
10.30-14.00 h. Laboratory
session:
T1: Disk diffusion
and microdilution.
T2: Preparation of agarose plugs for PFGE.
T5: Transference of ß-lactamase encoding
plasmids by conjugation.
T2: DNA extraction, REP-PCR and electrophoretic
analysis.
14.00-15.30 h.
Lunch.
15.30-17.30 h.
Laboratory session (cont.).
Resistance to antiretroviral agents by José Luis Pérez.
Day 3. Tuesday,
June 22
Mechanisms of resistance to quinolones in
Gram-positive bacteria by David Hooper.
09.45-10.15 h.
Objectives of the day.
10.15-14.00 h. Laboratory
session:
T5:
Evaluation of transconjugants and assay of their antimicrobial susceptibility
(E-test).
T2:
PFGE continued: restriction.
T2:
Staining and visualizing REP-PCR gel.
T3:
Isolation,electrophoretic analysis, preparation of competent cells and
electroporation of plasmids encoding DNA repair genes.
T4:
PCR amplification of gyrA and electrophoretic analysis.
14.00-15.30 h.
Lunch.
15.30-17.30 h.
Laboratory session (cont.)
Resistance and
virulence by Fernando Baquero.
8:30-9:30 h. Lecture.
Relevance of
horizontal gene transfer units in antimicrobial resistance by Fernando de la Cruz.
09.45-10.15 h. Objectives of the day.
10.15-14.00 h. Laboratory
session:
T4:
Visualization of gyrA amplicons, restriction, and agarose gel
electrophoresis.
T6:
Extraction of outer membrane proteins.
T5:
Antimicrobial susceptibility of transconjugants.
T3:
Evaluation of electroporation results.
T5:
Extraction for ß-lactamase assays (sonication).
T2:
PFGE continued: load and run gel.
13.30-14.30 h. Lecture.
Clinical versus
microbiological breakpoints by R. Cantón.
Day 5. Thursday, June 24
Hypermutable bacteria and antimicrobial resistance by Antonio Oliver.
09.45-10.15 h. Objectives of the day.
10.15-14.00 h.
Laboratory session:
T5:
Isoelectric focusing of ß-lactamase extracts.
T4:
Visualization of gyrA restriction.
T5:
Spectrophotometric assay of ß-lactamases.
T4:
RNA extraction.
T3:
Test hypermutation.
T2:
Interpretation of PFGE and REP-PCR. Discussion.
T6:
SDS-PAGE analysis of OMPs.
T5:
PCR amplification of ß-lactamase genes.
14.00-15.30 h.
Lunch.
15.30-17.30 h.
Laboratory session (cont.)
DNA microarrays by Jordi
Vila.
21.00 h Official dinner.
Day 6. Friday,
June 25
8:30-9:30 h. Lecture.
Animal models for evaluation
of antimicrobial resistance by Claude Carbon.
09.45-10.15 h. Objectives of the day.
10.15-14.00 h. Laboratory
session:
T4:
Analysis of mRNA expression.
T4:
Quinolones accumulation assays.
T6:
Interpretation of SDS-PAGE analysis of OMPs.
T3:
Discussion of results of mutation frequency.
14.00-15.30 h.
Lunch.
15.30-17.30 h. Laboratory session (cont.)
Environmental isolates as a source of resistance genes by Patrice Nordmann.
Day 7. Saturday,
June 26
8:30-9:30 h.
Lecture
Efflux
pumps inhibitors by Luis Martínez-Martínez.
09.45-10.15 h. Objectives of the day.
10.15-10:45 h. Laboratory
session:
T4:
Evaluation of mRNA expression.
10.45-11.30 h.
Questionnaire. Evaluation of the course.
11.30-13:00. Visit to Valldemosa.
Farewell Lunch:
On-the-Beach.