Transcript Headertitel

WP nr 6
Birgitta Henriques Normark, Sweden
Raquel Sa-Leao, Herminia de Lencastre, Portugal
Regine Hakenbeck, Germany
Chris Tang, UK
Staffan Normark, Sweden
Progress Year 4
Serotyping, molecular typing (MLST) and antibiotic resistance among LRTI isolates, GRACE. Totally
80+83 isolates. Clinical information needs to be correlated to the molecular characteristics of the strains.
Performed microarray analysis on clinical pneumococcal isolates belonging to different serotypes, sequence
types and of different invasive disease potential and we are studying the genetic relationsships between
different clonal complexes as determined with the sequenced based molecular typing method MLST. Gene
content variation among isolates of different eBURST groups may differ, potentially due to differences in
transformability. Furthermore, our data show that assumptions of similarity in large clonal complexes
should be done with caution. Pathogenicity islands studied in in vivo models .
Correlation of disease to genomic content of the bacteria by molecular typing and sequencing of invasive and
carriage isolates of the same serotype and clonal type
Impact of two different patterns of antibiotic consumption on pneumococcal resistance in a country with
widespread use of the seven-valent pneumococcal conjugate vaccine.
Highly penicillin-resistant multidrug-resistant pneumococcus-like strains colonizing children: genomic
characteristics and implications for surveillance.
The stability of a multiple antibiotic and high penicillin-resistant clone Spain 23F-1 has been analyzed by
comparing genome sequences of seven members of this clone Spain, South Africa, Hungary, Germany and
France. A similar approach has been started using three genomes of the Hu19A-6 clone. Two distinct
isolates have been sequenced which will be compared to the finished genome which is publicly available
A model of the respiratory epithelial barrier has been developed to examine the traversal of the
pneumococcus from the nasopharynx. Calu-3 cells are grown on semi-permeable membranes and become
polarised, develop tight junctions, and are impermeable to flourescent tracer.
23B
23F
31
33F
34
35B
35F
NT
EJ
23F
24F
28A
33F
35B
NT
PN?
EJ
18C
17F
16F
15C
15B
15A
14
11A
10A
9N
8
6B
6A
3
1
23A
0
22F
2
22F
4
21
6
19F
8
19F
10
19A
12
19A
14
17F
16F
12F
11A
9V
9N
9A
7F
6B
6B
6A
6
4
3
1
Progress Year 4
30
25
20
15
10
5
0
Progress Year 4
25
20
no of isolates
15
10
5
0
62
124
138
180
193
199
395
414
CC
433
460
1012
1994
others
not
finished
Deliverables Year 4
1. To create and maintain S. pneumoniae data base.
2. To collect pneumococcal isolates causing community-acquired LRTI in Europe.
3. To detect new and unusual antibiotic resistance profiles among S. pneumoniae causing
community-acquired LRTI in Europe.
4. To determine the serotype distribution of antibiotic susceptible and non-susceptible pneumococci
across Europe in community-acquired LRTI to inform vaccine design.
5. To monitor clonal spread of S. pneumoniae causing community-acquired LRTI in Europe
important for intervention regimens.
6. To correlate antibiotic resistance, virulence characteristics and pneumococcal genotype to disease
parameters such as severity of community-acquired LRTI aiming at optimise future treatment and
intervention strategies.
7. To monitor pathogenicity islands and horizontally acquired DNA among pneumococcal isolates
causing community-acquired LRTI by performing comparative genomics with microarray
technology with the aim of finding genes important for virulence and for antibiotic resistance
development.
8. To characterize in in vivo (mice) models the role of pathogenicity islands in pneumococcal
disease.
Milestones Year 4
WP6 – DL9: Month 42
Strain collection, frequency of resistance and serotypes among
pneumococcal isolates causing community-acquired LRTI during the
surveillance period
WP6 – DL10: Month 42
Molecular types of antibiotic susceptible and resistant pneumococcal
isolates identified in DL1 and DL2
WP6 – DL11: Month 48
Strain collection, frequency of resistance and serotypes among
pneumococcal isolates causing community-acquired LRTI during the
surveillance period
Obstacles (if any)
• Less isolates than anticipated
• Several optochin resistant
• Clinical information about the isolates needed to perform
correlations
Dissemination at meetings
INVITED ORAL TALKS AT CONFERENCES (SELECTED)
Sá-Leão, R. 2009. O ecossistema da nasofaringe (“The nasopharyngeal ecosystem”). Congresso Nacional de Pediatria (“National
Pediatrics Congress”). Tróia, Portugal. October 15-17, 2009
Henriques Normark, The Epidemiology of pneumococcus , Semmering, Austria, April 23-26 ,2009
Henriques Normark, Key note lecturer, ECCMID Helsinki, May 19,2009
Henriques Normark, Nobelsymposium Stockholm, June 11-13, 2009
Henriques Normark, Umeå Bacterial cellbiology and pathogenesis Invited speaker , Umeå, June 14-18, 2009
ABSTRACTS SELECTED FOR ORAL PRESENTATIONS IN INTERNATIONAL AND NATIONAL MEETINGS (SELECTED)
Frazão, N., R. Sá-Leão, and H. de Lencastre. 2009. Effect of a single dose of the 7-valent pneumococcal conjugate vaccine (PCV7)
on the composition of the nasopharyngeal flora. Abstr. D-O5, p. 59. In Programme and Abstract Book of the 9th European
Meeting on the Molecular Biology of the Pneumococcus. Bern, Switzerland. June 4 - 7, 2009.
Valente C., S. Nunes, R. Sá-Leão, and H. de Lencastre. 2009. Dynamics of serogroup 6 of Streptococcus pneumoniae during the
last decade in Portugal, including the recently described serotype 6C. Abstr. p. 29. In Programme and Abstract Book of the
9th European Meeting on the Molecular Biology of the Pneumococcus-Europneumo 2009. Bern, Switzerland, June 4 - 7 2009.
Simões A. S., R. Sá-Leão, S. Nunes, N. Frazão, A. Tavares, H. de Lencastre. Penicillin resistance caused by dissemination of
serotype 14 - clone Spain9V ST156 - in the vaccine era. Abstr. O494, p. 74. In Programme and Abstract Book of the 19th
European Congress of Clinical Microbiology and Infectious Diseases (ECCMID). Finland. May 16 - 19, 2009.
Nunes, S., R. Sá-Leão, A.S. Simões, N. Frazão, A. Brito-Avô, and H. de Lencastre. 2009. Colonization of Streptococcus
pneumoniae in the era of seven-valent conjugate pneumococcal vaccine. Abstr. ID 153. In Microbiotec’09 Oral Session S5:
Health and Pharmaceutical Biotechnology. Vilamoura, Portugal. November 28-30, 2009.
Sá-Leão, R., S. Nunes, N. Frazão, A.S. Simões, A. Brito-Avô3, and H. de Lencastre. 2009. “Impacto da vacina pneumocócica
conjugada sete-valente (PCV7) em colonização” Abstr. p. 7. In Programme and Abstract Book of the “XI Jornadas Nacionais
de Infecciologia Pediátrica – I Jornadas Lusófonas de Infecciologia Pediátrica”. Aveiro, Portugal. May 7 - 9, 2009.
SEVERAL POSTERS ON NATIONAL AND INTERNATIONAL CONFERENCES
Publication plan
PUBLISHED ORIGINAL ARTICLES IN SCIENTIFIC PEER-REVIEWED JOURNALS
Blomberg et al. Pattern of accessory regions and invasive disease potential in Streptococcus pneumoniae. J Infect Dis.
2009 Apr 1;199(7):1032-42.
Simões, A.S.†, R. Sá-Leão†, M.J. Eleveld, D.A. Tavares, J. Carriço, H.J. Bootsma, and P.W.M. Hermans. 2010. Highly
penicillin-resistant multidrug-resistant pneumococcus-like strains colonizing children: genomic characteristics and
implications for surveillance. J. Clin. Microbiol. 48:238-246.
Rodrigues, F.†, S. Nunes†, R. Sá-Leão, G. Gonçalves, L. Lemos, and H. de Lencastre. Streptococcus pneumoniae
nasopharyngeal carriage in children attending day care centers in the central region of Portugal, in the era of 7valent pneumococcal conjugate vaccine. Microb. Drug Resist. Microb. Drug Resist. 15:269-277.
Sá-Leão, R., S. Nunes, A. Brito-Avô, N. Frazão, A. Simões, M.I. Crisóstomo, A.C.S. Paulo, J. Saldanha, I. SantosSanches, and H. de Lencastre. 2009. Changes in pneumococcal serotypes and antibiotypes carried by vaccinated
and unvaccinated day-care center attendees in Portugal, a country with widespread use of the seven-valent
pneumococcal conjugate vaccine. Clin. Microbiol. Infect. 15: 1002-1007.
Nunes, S.†, C. Valente†, R. Sá-Leão, and H. De Lencastre. 2009. Secular trends and molecular epidemiology of the
recently described serotype 6C of Streptococcus pneumoniae. J. Clin. Microbiol. 47:472-474.
SUBMITTED MANUSCRIPTS
Blomberg C, Dagerhamn J, Browall S, Normark S, Henriques-Normark B. Gene content variation within clonal
complexes of Streptococcus pneumoniae Submitted
PLANNED
-LRTI PNEUMOCOCCAL ISOLATES (GRACE COLLECTION), CHARACTERISTICS CORRELATED TO CLINICAL INFORMATION
-GENOMIC COMPARISON AND CORRELATION TO CARRIAGE AND DISEASE