BACKGROUND: The aim of this study was to evaluate the presence of pilus islet 1 (PI-1) and to determine its clade type in pneumococcal isolates with reduced susceptibility to penicillin (penicillin non-susceptible pneumococci - PNSP) and/or resistant to macrolides isolated prior to and after the introduction of pneumococcal conjugate vaccines (PCVs) in the Czech Republic. METHODS: Clinical isolates of serotypes 9V (n = 68) and 19A (n = 89) were examined. Isolates were characterised by multilocus sequence typing (MLST). The presence of PI-1 was determined by screening for the sortase B, C, and D genes located within PI-1. In the presence of PI-1 pilus, clade types were classified by PCR. RESULTS: In the pre-PCV period (2000-2007), the prevalence of PNSP was 3.9% and 2.7% of isolates were resistant to erythromycin. During 2012-2015 (post-PCV period), the rates of PNSP remained stable (3.6%), but resistance to erythromycin increased to 8.3%. While in 2000-2007, resistance to antibiotics was associated mainly with serotype 9V, in 2012-2015, it was replaced by serotype 19A. PI-1 positive isolates were seen in both serotypes. All isolates (68) of serotype 9V belonged to the Spain9V-3 (CC156) clone and carried PI-1 of clade type I while 96.5% (56/58) of isolates of 19A serotype belonged to the Netherlands15B-37 (CC199) clone and carried PI-1 of clade type II. CONCLUSIONS: Both major antibiotic resistant clones carried PI-1, although they differ in the clade type. Thus the role of PI-1 should be evaluated in further studies and potentially considered in the spread of antibiotic resistant clones.

Centre for Epidemiology and Microbiology National Institute of Public Health Prague Czech Republic

Department of Clinical Microbiology Faculty of Medicine and University Hospital Charles University Hradec Kralove Czech Republic

Department of Infectious Diseases Faculty of Medicine Charles University Prague Budínova 2 180 81 Prague Czech Republic

Department of Infectious Parasitic and Tropical Diseases Na Bulovce Hospital Budínova 2 180 81 Prague Czech Republic

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$a Zemlickova, Helena $u Centre for Epidemiology and Microbiology, National Institute of Public Health, Prague, Czech Republic; Department of Clinical Microbiology, Faculty of Medicine and University Hospital, Charles University, Hradec Kralove, Czech Republic. Electronic address: hzemlickova@szu.cz

245    14

$a The association of pili with the emergence and replacement of the major antibiotic resistant pneumococcal clones / $c H. Zemlickova, V. Jakubu, M. Fridrichova, L. Malisova, . Martin Musilek, M. Trojanek

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$a BACKGROUND: The aim of this study was to evaluate the presence of pilus islet 1 (PI-1) and to determine its clade type in pneumococcal isolates with reduced susceptibility to penicillin (penicillin non-susceptible pneumococci - PNSP) and/or resistant to macrolides isolated prior to and after the introduction of pneumococcal conjugate vaccines (PCVs) in the Czech Republic. METHODS: Clinical isolates of serotypes 9V (n = 68) and 19A (n = 89) were examined. Isolates were characterised by multilocus sequence typing (MLST). The presence of PI-1 was determined by screening for the sortase B, C, and D genes located within PI-1. In the presence of PI-1 pilus, clade types were classified by PCR. RESULTS: In the pre-PCV period (2000-2007), the prevalence of PNSP was 3.9% and 2.7% of isolates were resistant to erythromycin. During 2012-2015 (post-PCV period), the rates of PNSP remained stable (3.6%), but resistance to erythromycin increased to 8.3%. While in 2000-2007, resistance to antibiotics was associated mainly with serotype 9V, in 2012-2015, it was replaced by serotype 19A. PI-1 positive isolates were seen in both serotypes. All isolates (68) of serotype 9V belonged to the Spain9V-3 (CC156) clone and carried PI-1 of clade type I while 96.5% (56/58) of isolates of 19A serotype belonged to the Netherlands15B-37 (CC199) clone and carried PI-1 of clade type II. CONCLUSIONS: Both major antibiotic resistant clones carried PI-1, although they differ in the clade type. Thus the role of PI-1 should be evaluated in further studies and potentially considered in the spread of antibiotic resistant clones.

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$a Jakubu, Vladislav $u Centre for Epidemiology and Microbiology, National Institute of Public Health, Prague, Czech Republic

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$a Fridrichova, Marta $u Centre for Epidemiology and Microbiology, National Institute of Public Health, Prague, Czech Republic

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$a Malisova, Lucia $u Centre for Epidemiology and Microbiology, National Institute of Public Health, Prague, Czech Republic

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$a Martin Musilek, $u Centre for Epidemiology and Microbiology, National Institute of Public Health, Prague, Czech Republic

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$a Trojanek, Milan $u Department of Infectious Diseases, Faculty of Medicine, Charles University in Prague, Budínova 2, 180 81 Prague, Czech Republic; Department of Infectious, Parasitic and Tropical Diseases, Na Bulovce Hospital, Budínova 2, 180 81 Prague, Czech Republic

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