Characteristics of Streptococcus pneumoniae serotype 19A isolates from children in the pre and post Conjugate Vaccine Era. Single center experience 1986-2015.

BACKGROUND: The present study assessed the prevalence and characteristics of S. pneumoniae serotype 19A isolates from children with pneumococcal disease (PD), before and since introduction of pneumococcal conjugate vaccines (PCVs) in Greece. METHODS: S. pneumoniae isolates collected at one large pediatric hospital between 1986 and 2015 were serotyped by the Quellung reaction and MICs determined by Etest. Alterations of pbp genes and the presence of mefA, mefE, ermB genes were detected by polymerase chain reaction. Genotypes were assessed by multilocus sequence typing (MLST). RESULTS: Among 1875 isolates, 210 (11.2%) belonged to serotype 19A. The prevalence of PD caused by serotype 19A increased from 4.6% in the pre-PCV7 years (1986-2005) to 19.6% in the post-PCV7 years (2006-2010), peaking at 27% in 2009 (p < 0.001, 95% CI; 2.0, 18.2) with a significant upward trend (p = 0.04, 95% CI; 1.02, 12.66). Following the introduction of PCV13 in 2010, the rate decreased from 22% in 2011 to 11.4% in 2015 (p = 0.08, 95% CI; 0.92, 5.1) with a downward trend of borderline significance (p = 0.05, 95% CI; -6.8, 0.04). The multidrug resistant (MDR) serotype 19A isolates increased from 10.6% in 1986-2005 to 21.2% in 2006-2010 and to 71.8% in 2011-2015 (P < 0.001). Alterations in pbp genes were detected in all penicillin non-susceptible isolates. Of 110 erythromycin resistant isolates, 21 contained the mefE gene, 36 the ermB and 53 both the mefE and ermB genes. MLST analysis of 142 isolates revealed four dominant clonal complexes (CC); CC320, CC172, CC276 and CC199. The majority of CC320 and CC276 isolates displayed MDR phenotypes. CONCLUSION: PD caused by serotype 19A increased significantly after the introduction of PCV7 followed by a decline after PCV13 use. The vast majority of persisting 19A isolates was MDR. Surveillance studies are necessary to monitor the changes in the pneumococcal population.

Molecular analysis of Streptococcus pyogenes macrolide resistance of paediatric isolates during a 7 year period (2007-13).

OBJECTIVES: The molecular characterization of paediatric group A Streptococcus (GAS) isolates regarding macrolide resistance and relevant emm types in Athens, Greece. METHODS: Pharyngeal and non-pharyngeal GAS isolates were collected during a 7 year period (2007-13) and examined for antibiotic susceptibility, macrolide resistance genes [mef(A), erm(A) and erm(B)] and relevant emm types. RESULTS: Overall, 20.4% (270/1324) of GAS isolates were resistant to macrolides. The macrolide resistance rate varied during the study period with a maximum rate observed in 2008 (29.57%) and a minimum rate observed in 2013 (10.95%) (P value for trend = 0.007). During the same period, consumption of macrolides was gradually reduced by 56.6%. No difference was observed in macrolide resistance between pharyngeal and non-pharyngeal isolates (P = 0.7). Among macrolide-resistant isolates, mef(A) was detected in 87 (32.2%), erm(A) in 136 (50.4%), erm(B) in 44 (16.3%) and both mef(A) and erm(A) in 3 (1.1%) isolates. The most prevalent emm types among macrolide-resistant isolates were emm77 (31.5%), emm4 (18.1%) and emm12 (10.7%). Ten emm types (77, 4, 12, 28, 1, 22, 11, 2, 44 and 89) accounted for 90.3% of macrolide-resistant isolates. emm types 4, 22, 44 and 77 were more prevalent in macrolide-resistant compared with macrolide-susceptible isolates, whereas emm types 1, 3, 5, 6, 75 and 89 were more prevalent in macrolide-susceptible compared with macrolide-resistant isolates. CONCLUSIONS: GAS macrolide resistance remained significant in our area during the study period. A substantial decline in the resistance rate was observed in the last year of the study, which may be related to reduced consumption of macrolides.

Pneumococcal Mastoiditis in Children Before and After the Introduction of Conjugate Pneumococcal Vaccines.

OBJECTIVES: To determine whether serotype distribution and antibiotic resistance of Streptococcus pneumoniae acute mastoiditis (AM) in children have changed in the post pneumococcal conjugate vaccines (PCVs) era. METHODS: Medical records of pneumococcal AM cases, in a tertiary pediatric hospital were reviewed from January 1999 to December 2014. S. pneumoniae isolates were serotyped using the quellung reaction and tested for antibiotic susceptibility by E-test and for macrolide resistance genes by polymerase chain reaction. RESULTS: Among 334 children with AM, S. pneumoniae was isolated from 89 (26.6%) with median age 22 months (interquartile range: 12-30 months). S. pneumoniae was recovered from ear fluid (58%), mastoid specimens (35.2%) and blood (6.8%). Resistance to penicillin, erythromycin and clindamycin was 12.4%, 49.4% and 18%, respectively. Distribution of pneumococcal serotypes before (1999-2005), after the introduction of PCV7 (2006-2010) and after PCV13 (2011-2014) was found: for the PCV7 serotypes 81%, 25% and 0% (P < 0.0001), for PCV13 additional serotypes 16.3%, 70.8% and 63.6% (P < 0.0001) and for non-PCV serotypes 2.3%, 4.1% and 36.3% (P = 0.0002), respectively. Significant increase was detected for the serotype 19A after PCV7, and this trend was not changed after PCV13 (2.3%, 50% and 50%, respectively; P < 0.0001). A significant proportion of resistant isolates to penicillin (54.5%) and erythromycin (34.8%) was identified as 19A. CONCLUSIONS: After the introduction of PCV7, a significant increase of serotype 19A and replacement of PCVs serotypes was identified. After PCV13, the overall proportion of pneumococcal mastoiditis and the incidence of serotype 19A were not significantly declined. A significant proportion of resistant isolates to penicillin and erythromycin is attributed to serotype 19A.

Macrolide resistance in Streptococcus pyogenes: prevalence, resistance determinants, and emm types.

To investigate the antimicrobial resistance trends and the distribution of emm types of group A streptococci (GAS), we examined 1160 clinical isolates of GAS collected between 2003 and 2006. Susceptibilities to commonly used antimicrobial agents were determined by Etest, and macrolide resistance genes were detected by polymerase chain reaction (PCR). GAS isolates were typed by polymerase chain reaction PCR and sequencing of emm gene. The rates of resistance to erythromycin (ERY), clindamycin, azithromycin, tetracycline, and chloramphenicol were 14.9%, 1.4%, 14.9%, 18.9%, 0.6%, respectively. None of the isolates exhibited resistance to penicillin, ceftriaxone, linezolid, moxifloxacin, rifampicin, or vancomycin. Macrolide resistance increased from 12.1% in 2003 to 18.8% in 2006 (P = 0.02). Of 173 ERY-resistant GAS isolates, 93 (53.7%) harbored the mefA gene, 70 (40.4%) the ermA, and 10 (5.8%) the ermB. Eighty percent of the observed emm types are covered by the proposed 26-valent GAS vaccine. Among 173 ERY-resistant isolates, the predominant emm types were 12 (19.5%), 77 (17.9%), and 4 (16.8%), and among 770 ERY-susceptible isolates, the predominant types were 1 (18.8%), 12 (17.5%), 28 (13.8%). The observed antimicrobial resistance trends and the distribution of specific emm types have implications in guiding empiric therapy and in developing vaccine strategies to prevent GAS infections.