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Decreased Carriage and Genetic Shifts in the Streptococcus pneumoniae Population After Changing the Seven-valent to the Thirteen-valent Pneumococcal Vaccine in Norway

Steens, Anneke MSc*; Caugant, Dominique A. Fil.dr*†; Aaberge, Ingeborg S. MD, PhD*; Vestrheim, Didrik F. MD, PhD*‡

The Pediatric Infectious Disease Journal: August 2015 - Volume 34 - Issue 8 - p 875–883
doi: 10.1097/INF.0000000000000751
Vaccine Reports
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SDC

Background: Shifts in the pneumococcal population colonizing healthy children are expected after switching from a 7-valent pneumococcal conjugate vaccine (PCV7) to a 13-valent (PCV13) in the childhood immunization program. We assessed effects of the switch by comparing pneumococcal carriage and serotype and genetic diversity of pneumococci carried by children in the PCV13-era with those carried in the prevaccination-era and PCV7-era.

Methods: Nasopharyngeal swabs were obtained in autumn 2013 from children attending day-care centers (874 swabs, 583 isolates). Serotyping, multilocus sequence typing and antimicrobial susceptibility testing were performed on all isolates. Results were compared with samples from 2006 (610 swabs, 538 isolates) and 2008 (600 swabs, 562 isolates).

Results: The carriage prevalence in 2013 was 62 of 100 children (95% confidence intervals: 58–66), a significant decrease from 2006 and 2008. PCV13 serotypes accounted for 7% of isolates in 2013. Non-PCV13 prevalence increased from 2006 to 2008 [prevalence ratio: 1.73 (1.40–2.15)] but remained stable in 2013 [0.99 (0.88–1.12)]. Still, non-PCV13 serotypes 21, 23B, 23A and 22F had increased. In 2013, the serotype and genetic diversity had decreased slightly, and distinct serotype and genetic profiles clustered more within day-care centers compared with the earlier samples. Serotype switch was uncommon. Overall, antimicrobial resistance was limited.

Conclusions: Carriage of PCV13 serotypes has decreased without a coinciding increase in non-PCV13 serotypes. The serotype and genetic shifts among non-PCV13 serotypes suggest that a new equilibrium has not yet been reached. As the few non-PCV13 serotypes that increased have generally a lower invasive capacity than vaccine serotypes, direct and indirect protection of PCV13 on invasive pneumococcal disease can be expected to continue.

Supplemental Digital Content is available in the text.

From the *Division of Infectious Disease Control, Department of Bacteriology and Immunology, Norwegian Institute of Public Health, Oslo, Norway; Faculty of Medicine, University of Oslo, Oslo, Norway; and European Programme for Public Health Microbiology Training (EUPHEM), European Centre for Disease Prevention and Control (ECDC), Stockholm, Sweden.

Accepted for publication February 3, 2105.

The authors have no conflicts of interest or funding to disclose.

Supplemental digital content is available for this article. Direct URL citations appear in the printed text and are provided in the HTML and PDF versions of this article on the journal’s website (www.pidj.com).

Address of correspondence: Anneke Steens, MSc, Department of Bacteriology and Immunology, Norwegian Institute of Public Health, P.O. Box 4404, Nydalen NO-0403, Oslo, Norway. E-mail: anneke.steens@fhi.no.

Streptococcus pneumoniae (the pneumococcus) commonly colonizes the human nasopharyngeal epithelium.1 Children carry pneumococci more often and for longer episodes than adults2,3 and are considered the main reservoir.4,5 Pneumococcal carriage can lead to disease, such as pneumonia and invasive pneumococcal disease (IPD).6 Pneumococcal conjugate vaccines (PCV) have been widely implemented in childhood immunization programs and have been shown to effectively prevent both IPD7,8 and asymptomatic carriage3 of vaccine serotypes (VT). As PCV affect VT carriage, the composition of the pneumococcal population changes after vaccine implementation. Removal of VT through vaccination opens nasopharyngeal niches for colonization by non-vaccine serotypes (NVT). Changes in the colonizing population can occur through expansion of preexisting clones, introduction of clones not previously identified in the country or serotype switching; a mechanism to escape the vaccine-induced immune response.9 Depending on the invasive capacity of the replacing serotypes,10–12 increased incidence of disease caused by NVT may occur. Serotype replacement has been observed after the introduction of a 7-valent vaccine (PCV7) in childhood immunization programs,13–16 but overall effects of PCV7 were positive and included direct protection of immunized children as well as indirect protection of non-targeted age groups.13,16,17 A new equilibrium within the pneumococcal population was reached a couple of years after vaccine introduction.18

A 13-valent PCV (PCV13) has been available and widely used in immunization programs since 2010. Early results suggest that PCV13 also provides indirect protection,19–21 and that serotype replacement in IPD is occurring to a lesser degree than after PCV7 introduction.19,20,22 The net effect in the longer run is not yet known and will depend on the new equilibrium that will be reached in the pneumococcal population.

To understand and predict postvaccination changes in IPD, the dynamics in serotype and genotype diversity of carried strains should be studied. We, therefore, assessed vaccine-induced changes in carriage prevalence and in serotype and genetic diversity of pneumococci carried by children, the main transmitters. Results from a cross-sectional carriage study conducted almost 2 years after switching from PCV7 to PCV13 were compared with similar samples taken before mass vaccination with PCV723 and almost 2 years after PCV7 introduction.24

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MATERIALS AND METHODS

Study Population

Children attending day-care centers (DCC) in and around Oslo, the capital of Norway, were included in the study. We used DCC as primary sampling unit, and invited all children per DCC. We selected communal and private DCC, excluding small family DCC, short-term DCC or DCC where no Norwegian is spoken. In Norway, 90% of the children aged 1–5 years attend DCC.25 Pneumococcal immunization is administered in a 2 + 1 dose schedule at 3, 5 and 12 months. PCV7 was used from July 2006 and replaced by PCV13 in April 2011.

The study was conducted in accordance to the principles of the Declaration of Helsinki and was approved by the Regional Committee for Medical Research Ethics, South-Eastern Norway. The parents/guardians gave informed consent before including their child in the study.

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Data and Sample Collection

Data and samples were collected in autumn 2013. Information was obtained from parents/guardians using a questionnaire on demographics, household properties, risk factors for carriage, number of PCV doses received, history of upper respiratory tract infections and antimicrobial use in the 3 months preceding sampling (Table 1). As the switch to PCV13 occurred nationally on April 11, 2011, the type of vaccine used (PCV7, combination of PCV7+PCV13, or PCV13) was assigned based on the date of birth, assuming that the vaccine doses were administered according to schedule. This is, however, an approximation, as about 8% of Norwegian children receive their first PCV more than 1 month after recommended schedule and the second and third doses are delayed in 19% and 22% of the children, respectively.26

TABLE 1

TABLE 1

Nasopharyngeal swabs (E-swabs, Copan, Brescia, Italy) were collected according to standard procedures, as previously described.24 After sampling, swabs were inoculated in enrichment broth [Statens Serum Institut (SSI), Copenhagen, Denmark]. Within 4 hours the swabs were transported to the laboratory and plated on gentamicin blood agar. Swabs in enrichment cultures and blood-agar with 5 µg gentamicin supplemented per ml plates were incubated overnight at 35°C, with 5% CO2.

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Bacterial Identification and Serotyping

Pneumococci were identified as described.24 In short, serotyping was performed from enrichment cultures using a commercial latex agglutination kit (Pneumotest-Latex kit; SSI). Confirmation and factor typing were performed by the capsular reaction test (Quellung reaction) using specific antisera (SSI). All morphologically different pneumococcal colonies per sample were typed. If additional serogroups/serotypes were identified in the agglutination test, up to 16 colonies were isolated in the attempt to identify the different strains.

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Antimicrobial Susceptibility Testing

The minimum inhibitory concentration (MIC) was determined for penicillin G, ceftriaxone, erythromycin, clindamycin, tetracycline and trimethoprim/sulfamethoxazole using antimicrobial gradient strips (Etest, Biomérieux, Paris, France). Isolates were characterized as susceptible, intermediate or resistant using EUCAST breakpoints, v3.1.27

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Genetic Analysis

Genomic DNA was prepared as previously described.23 Multilocus sequence typing (MLST) of all isolates was performed on basis of 7 housekeeping genes.28 Sequence types (ST) were determined using Seqscape v2.5 and Sequencher v4.8 with the MLST-database (http://pubmlst.org/spneumoniae/). Novel alleles and ST were assigned designations by the database curator.

A clone was defined as isolates belonging to the same ST. Clonal complexes (CC; 6 of 7 alleles shared with other ST identified in our dataset) and singletons (≤5 of 7 alleles in common with other ST) were determined using eBURST (http://eburst.mlst.net/v3/enter_data/). A comparative eBURST analysis using the entire online MLST database was also run. If the predicted founder of our CC was different from the predicted founder of the entire database (international CC), both names are provided in the text.

To identify whether ST/serotype combinations were novel, and thus could have arisen from serotype switching, we searched whether the combinations had been described before using http://pubmlst.org/perl/bigsdb/bigsdb.pl?db=pubmlst_spneumoniae_isolates&page=listQuery. Clonal expansion and introduction of clones not previously identified in Norway were defined based on their occurrence in 2006 and/or 2008.

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Data Analysis

Age group and VT/NVT-specific prevalences were determined per 100 children (number of children with serotype/number of children in that stratum × 100). Serotypes were categorized as PCV7 (4, 6B, 9V, 14, 18C, 19F and 23F), additional serotypes in PCV13 (PCV13–7; 1, 3, 5, 6A, 7F, 19A) and non-PCV13 (all other serotypes). We corrected analyses for clustering of children in DCC using the survey command in Stata. Results are presented with 95% confidence intervals (95% CIs) in parentheses.

The results of the 2013 sample (PCV13 era) were compared with samples obtained in 2006 (prevaccination) and 2008 (PCV7 era). Parts of the data of the 2006 and 2008 samples have been published previously.23,24 The design of all these samples was similar. Prevalence ratios (PR) were estimated using a Poisson model, with the incidence-rate ratio command in Stata. Assumptions were tested using Pearson’s goodness-of-fit. Statistical significance (P < 0.05) of changes in serotype-specific prevalence was determined using the incidence-rate ratio calculator.

The variability in carriage per DCC per year was expressed by its range; significant differences were determined by permutation test, with 5000 randomly sampled permutations. Simpson’s index of diversity (D-index)29 was used to express the diversity of serotypes and ST; D-index takes into account the total number of isolates, the number of isolates with a certain serotype or ST and the number of distinct serotypes or ST.18 The 95% CIs for the D-indexes were determined using bootstrap (5000×). We used Venn diagrams to describe the percentage of STs that persisted versus those that were only found in a certain year. The percentage of ST was calculated both as an unweighted percentage (ie, number of distinct ST/overall number of distinct ST found in the 3 years) and a weighted percentage (number of isolates comprising the distinct ST/overall number of isolates in the 3 years).

Analyses were performed in Excel 2010 and Stata 12.

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RESULTS

The 2013 Sample (PCV13 Era)

Overall 874 children, attending 40 DCC, were included (Table 1). The children were between 10 and 77 months old. Ninety-nine percent (95% CI: 98–99) had received pneumococcal vaccination (n = 859; for 5 children, vaccine status was missing). Assuming vaccination on schedule, 120 [14% (12–17)] of the children had received a combination of PCV7 and PCV13 and 246 [29% (25–33)] PCV13 only.

Sixty-two percent (58–66) of children carried pneumococci (n = 544; Fig. 1); 38 [7% (5–10)] of the carriers harbored more than 1 serotype. The percentage of carriers differed substantially between DCC, ranging from 20% to 89%, and by age groups (Table 2). The highest carriage prevalence was observed among children younger than 24 months [73 of 100 children (65–80)] and the lowest prevalence [51 of 100 (42–60)] in the ≥60 months old.

TABLE 2

TABLE 2

FIGURE 1

FIGURE 1

The 583 isolates belonged to 28 serotypes. Twenty-five isolates [4% (3–7)] were PCV7-serotypes (19F, 18C), and 40 isolates [7% (4–11)] had a serotype included in PCV13 but not in PCV7 (19A, 3, 7F, 6A). The overall prevalence of PCV13 serotypes was 7 of 100 children (5–10). The 5 most prevalent serotypes were 15B/C, 21, 35F, 23B and 23A (Fig. 2); 49% of isolates belonged to these non-PCV13 serotypes.

FIGURE 2

FIGURE 2

In 2013, 93 ST were identified, of which 10 constituted 50% of the isolates. Twenty ST (22% of ST; 7% of isolates) were novel to the MLST database. Fifty-eight ST belonged to 19 CC, and there were 35 singletons. The dominating CC were CC439, CC199 and CC177/international CC193; see (Supplemental Digital Content 1, http://links.lww.com/INF/C139) for an overview of all serotypes, ST and CC found in the 3 samples. Multiple capsular serotypes were expressed by ST162, ST62, ST177 and ST199; the ST/serotype combinations ST162/31, ST62/23A and ST177/24F had not yet been reported in the online MLST database. In addition, 5 other novel ST/serotype combinations were observed: ST1609/23A, ST945/23B, ST2154/15BC, ST4310/6C and ST2958/11A. These novel ST/serotype combinations accounted for 17 isolates in total; none of the combinations exceeded 7 isolates.

Decreased susceptibility (intermediate or full resistance) to at least 1 antimicrobial agent was found in 90 isolates [15% (11–21) of isolates]; Table 3. Thirty-nine isolates [7% (4–11)] were intermediate resistant to penicillin (MIC > 0.064 mg/L, the meningitis breakpoint for penicillin resistance); 36% of these were serotype 23B and belonged to CC1349/international CC156. Twenty-two isolates [4% (2–6)] were resistant to erythromycin. Coresistance to erythromycin, clindamycin and tetracycline was found in 18 isolates [3% (2–6)]. These coresistant isolates belonged to ST/serotypes ST179/19F, ST63/15A, ST344/NT and ST4310/6C. Resistance to trimethoprim/sulfamethoxazole only was found in 29 isolates [5% (3–8)], mainly among ST162/24F and ST193/21. Strains with decreased susceptibility were recovered from 70% of the DCC.

TABLE 3

TABLE 3

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Comparing the PCV13 Sample with the Prevaccination and PCV7 Samples

The 2013 sample did not differ from the 2006 and 2008 samples regarding the main demographics and potential risk factors for carriage (Table 1). By design, the vaccine uptake differed between samples. The uptake of the “new vaccine” did not differ between 2008 and 2013: 40% (35–44) for PCV7 versus 42% (38–46) for at least 1 immunization with PCV13. In 2013, children were reported to have suffered from upper respiratory tract infections in the past 3 months slightly more often than in 2008 [2013: 9% (7–12); 2008: 6% (4–8)], and recent antimicrobial use was higher [2013: 10% (8–12); 2008: 5% (4–8)].

While the overall carriage prevalence remained constant around 80% between 2006 and 2008 [PR: 1.03 (0.97–1.11)], the prevalence in 2013 was significantly lower [PR: 0.77 (0.72–0.84); Fig. 1]. Identification of multiple serotypes within individual children was less frequent in 2013 than in 2006 and 2008 (Table 4). The percentage of carriers among DCC varied more in 2013 than in earlier years (P = 0.028; Table 4). The prevalence of each PCV13 serotype was more than halved from 2008 to 2013, except for serotype 19A (Fig. 2). Five of the PCV7 serotypes were absent in 2013. The decrease of overall carriage and of carriage of PCV13 serotypes was observed for all age groups (Table 2). While carriage of non-PCV13 serotypes increased substantially from 2006 to 2008 [PR 1.73 (1.40–2.15)], no further increase was seen in 2013 [PR = 0.99 (0.88–1.12)]. The prevalence of non-PCV13 serotypes 21, 22F, 23A and 23B increased significantly compared with earlier years (called: the expanding NVT), with serotype 23B not being observed in 2006 or 2008. The prevalence of serotypes 21 and 22F already had increased before PCV13 introduction, whereas non-PCV13 serotypes 8, 15B/C, 16F, 33F, 35B and NT had significantly decreased.

TABLE 4

TABLE 4

Decreased susceptibility to any of the tested antimicrobial agents did not significantly differ over the years [2006: 16% (10–24); 2008: 13% (8–19); 2013: 15% (11–21)], but in 2013 a larger percentage of isolates was intermediate resistant to penicillin [2006: 2% (1–4); 2008: 2% (0–5); 2013: 7% (4–11)]. None of the isolates were penicillin resistant (MIC > 2 mg/L). The percentage of isolates with intermediate resistance to penicillin was higher among the expanding NVT [11% (5–22)] than among other isolates [5% (2–9); χ2P value = 0.009]. Eighty-two percent of isolates that were intermediate resistant to penicillin in 2013 belonged to ST that had not been observed in 2006 or 2008.

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Serotype and Genetic Diversity

As the pneumococcal population increased in diversity after PCV7 introduction coinciding serotype replacement,18 we investigated the diversity after the switch to PCV13. The number of serotypes decreased to 28 in 2013, from 32 in 2006 and 34 in 2008, and the D-index for the 2013 sample was lowest (though not significantly different from the 2006 level; Table 4). Furthermore, serotypes clustered more within DCC in 2013, as indicated by the lower median D-index per DCC, but the diversity was only slightly lower than in 2006. The number of ST in 2013 was the same as in 2008, but a smaller number of ST constituted 50% of isolates. Also for ST, the D-index was lowest for the 2013 sample.

Of the 197 ST identified overall in the 3 samples, only 28 ST (14%) were found in all 3 samples (Fig. 3A); these constituted 53% of all isolates (Fig. 3B).

FIGURE 3

FIGURE 3

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Changes in Genotypic Composition of Expanding NVTs and Persisting VTs

To disentangle underlying mechanisms leading to the increase of certain non-PCV13 serotypes and persistence of some PCV13 serotypes, changes in ST distribution were analyzed. The increases in non-PCV13 serotypes were mainly caused by clonal expansion (48% of the expanding NVT isolates) and introduction of clones not previously identified in the Norwegian carriage samples (47%). Five percent of expanding NVT isolates expressed a new ST/serotype combination. For all other serotypes found in 2013, 69% were clones existing in Norway, 29% were not previously identified in Norway and 2% presented a new ST/serotype combination. Serotype 21 increased through expansion of ST1877, the introduction of ST432 and appearance of 3 novel ST (ST9413, ST9414 and ST9522). The increase of serotypes 23A and 23B was caused mainly by introduction of clones; 77% of serotype 23A isolates in 2013 belonged to ST that were not observed in previous samples, including 2 novel ST belonging to CC439. The 23B isolates mainly belonged to the CC439 and the penicillin-intermediate resistant CC1349 (part of the international CC156). Interestingly, ST439 was observed in 2006 and 2008 with a 23A capsule and in 2013 with a 23B capsule. The serotype 22F increase resulted mainly from expansion of 1 clone, ST433.

Six PCV13 serotypes persisted in 2013. Among these, fewer ST per serotype were found, indicating decreased genetic diversity. Serotype 19F was dominated by isolates belonging to CC177/international CC193, mainly ST177 and ST179. The ST179/19F isolates were coresistant to erythromycin, clindamycin and tetracycline, with an increased prevalence in 2013, following a decrease from 2006 to 2008. The prevalence of ST177/19F decreased, while 2 ST177 isolates with a 24F capsule were observed in 2013. Only a single clone of serotype 3 (ST180) persisted in 2013, but with decreased prevalence. The nonsignificant increase in serotype 19A was mainly because of an increase in the antimicrobial susceptible ST199. PCV13 serotypes 18C, 6A and 7F persisted in 2013 with only a few isolates, but introduction of ST not previously identified in Norway was observed for these serotypes. ST162, associated with serotype 9V in 2006, appeared in 2013 with other capsules; 24F (majority), 15BC and 31.

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DISCUSSION

Our study revealed a decreased carriage prevalence of PCV13 serotypes after switching from PCV7 to PCV13 in the childhood immunization program, confirming its preventive effect on VT carriage.11,30–32 This decrease has already been reflected in direct and indirect protection against VT IPD in Norway.19 In contrast to what happened after PCV7 introduction,24 this decrease occurred without a coinciding increase in the overall prevalence of non-PCV13 carriage. Still, serotype and genetic shifts occurred among non-PCV13 serotypes. Decreased diversity and increased clustering of carriage and of clones within DCC were observed, indicating that a new equilibrium within the pneumococcal population might not yet have been reached, and that serotype replacement in carriage may be delayed instead of absent.

An overall decreased prevalence of pneumococcal carriage after switching to PCV13 has not been described in other settings,11,30–32 except Canada.33 The percentage of carriers found in our study was still higher compared with what has been found elsewhere. We serotyped from enrichment broth using a latex agglutination kit and the capsular reaction test, which permits higher detection of carriage and cocolonization with multiple serotypes compared with studies using conventional culture methods. The high carriage prevalence may, furthermore, be related to the low antibiotic use in Norway, as well as to the fact that the study was performed among children in DCC, where acquisition of pneumococci is frequent.34,35 With a high prevalence, a decrease might be easier to detect. Although the study was performed in and around the capital, we expect the results to be representative for Norway, as the majority of Norwegian children attend DCC and vaccine uptake is high in the entire country. We compared cross-sectional samples to investigate vaccine-related changes over time using samples obtained from a population with similar age distribution, in the same season, with similar sampling and laboratory techniques. The decreased carriage (observed after switching to PCV13) is, therefore, unlikely the result of our study design. A lower acquisition frequency, shorter duration of carriage episodes and/or lower pneumococcal density might have caused the decreased carriage prevalence. The decreased number of serotypes and the lower number of children carrying multiple serotypes indicate decreased frequency of pneumococcal acquisition, also because the currently dominant serotypes are known to be carried for relatively long periods of time.36

The unchanged prevalence of non-PCV13 carriage after switching to PCV13 indicates much more limited serotype replacement compared with what was seen after PCV7 introduction.3,14,24,37 A carriage study in Massachusetts performed shortly after switching to PCV13 did not either find indication for serotype replacement.38 It needs to be determined whether the absence of serotype replacement is permanent or simply delayed. The slight increase in clustering of serotypes and ST within DCC and the larger range of percentage of carriers per DCC may indicate that replacement occurs at a slower pace, as more similar pneumococcal populations in different DCC would be expected at equilibrium. Furthermore, there was a trend of lower serotype and genetic diversity in 2013. After universal vaccine implementation, the diversity in the pneumococcal population shortly decreases, as vaccination removes VT. When vacated niches become colonized, the diversity temporarily increases, until a new equilibrium has been reached.18 The slightly decreased diversity after switching to PCV13 may, therefore, indicate that all niches vacated by PCV13 have not yet been colonized, and that the redistribution of serotypes is delayed. Such delayed response may be explained by different properties of the PCV7 serotypes compared with the additional serotypes in PCV13, with serotypes 1 and 5 being rarely carried. Replacing serotypes might, by definition, be worse colonizers; they would otherwise have been the dominant carried serotypes already.

Despite the unchanged prevalence of non-PCV13 carriage, some non-PCV13 serotypes increased, especially serotype 23B. Increase of serotypes 23B and 22F among IPD cases aged 5 years or older in Norway was observed immediately after shifting to PCV13.19 In Canada33 and UK,22 increases in serotype 23B carriage have been observed among children. Interestingly, the main 23B clone found in this study was a ST that, in Norway, previously only carried 23A capsule. ST439/23B has been described before in the MLST database, suggesting introduction of an existing clone rather than serotype switching in Norway. The same probably accounts for ST162, which previously expressed the 9V capsule, but in 2013 mainly expressed the 24F capsule. Of the 8 novel ST/serotype combinations, ST1609/23A, ST945/23B, ST2154/15BC, ST4310/6C, ST177/24F and possibly ST162/31 went from a VT to a NVT, which may reflect vaccine escape through serotype switching. As the prevalence of these ST was low (≤0.8 of 100 children), the selective advantage seems to be limited. In 2008, only 1 ST was found with a novel ST/serotype combination, which had changed from a VT to a NVT (ST200/15BC24). ST200/15BC was not found in 2013. Overall, increases in carriage of non-PCV13 serotypes resulted from expansion of existing clones and introduction of clones that had not previously been identified in Norway, with limited serotype switching. This resembles what was observed the first years after PCV7 introduction.39,40 Approximately half of the isolates of the expanding NVTs was constituted of clones previously not identified in Norway, compared with less than one third of isolates of all other serotypes.

PCV13 carriage drastically decreased, but PCV13 serotypes 19F, 19A and 3 were still present. Worldwide, serotype 19A has been reported as a main replacing serotype after PCV7 introduction,13,15 and ST199 was the main replacing clone in Norway.41 As no carriage data from just before the switch to PCV13 are available, we do not know whether 19A carriage continued to increase after 2008 and started to decrease after switching to PCV13, as seen for IPD,19 or whether the 19A prevalence has remained stable. Several studies suggest that the effectiveness of PCV13 for serotype 3 is lower compared with other serotypes.42–47 Although serotype 3 was still present in 2013, we observed a decrease in its prevalence and its genetic diversity, suggesting some effectiveness of PCV13 on preventing carriage of serotype 3. The persistence of 19F may be because of advantages caused by antimicrobial resistance, as the antimicrobial susceptible ST177 decreased in prevalence while the resistant ST179 increased.

Overall antimicrobial resistance is limited among pneumococci in Norway. Still, the increase of intermediate resistance to penicillin (specifically among the expanding NVT) and presence of coresistant strains are worrying and warrants further monitoring. Carriage of resistant strains was not associated with the use of antimicrobials (data not shown), and although the percentage of children with reported use of antimicrobials in the 3 months preceding sampling had increased in 2013, no country-wide increase in use of penicillin or erythromycin has been registered in the Norwegian prescription database.48 In Canada, a similar initial decrease in antimicrobial usage after PCV7 introduction was followed by a small increase after switching to PCV13.33

In conclusion, overall pneumococcal carriage among children decreased after switching to PCV13, as a result of decreased VT carriage without a coinciding increase of non-PCV13 carriage. However, the observed serotype and genetic shifts among non-PCV13 serotypes suggest that a new equilibrium has not yet been reached, and increases in NVT carriage may still occur. Fortunately, as the few non-PCV13 serotypes that increased have generally a lower invasive capacity than VT, except for serotype 22F,10–12 direct and indirect effects of PCV13 on IPD can be expected to continue. Serotype switching, which could indicate escape of the vaccine-induced immune response, rarely occurred.

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ACKNOWLEDGMENTS

We are grateful to the children and their parents that participated in this study and thank the DCC workers for their support. We are thankful to Ingvild Essén and Line Tyskø for the collection of nasopharyngeal swabs. We acknowledge Anne Ramstad Alme, Gunnhild Rødal, Lene Haakensen and Torill Alvestad for the laboratory analyses and Hang Thi Ngoc Le for making all agar plates. We thank Richard White for statistical advice.

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REFERENCES

1. Bogaert D, Keijser B, Huse S, et al. Variability and diversity of nasopharyngeal microbiota in children: a metagenomic analysis. PLoS One. 2011;6:e17035
2. Melegaro A, Gay NJ, Medley GF.. Estimating the transmission parameters of pneumococcal carriage in households. Epidemiol Infect. 2004;132:433–441
3. Spijkerman J, van Gils EJ, Veenhoven RH, et al. Carriage of Streptococcus pneumoniae 3 years after start of vaccination program, the Netherlands. Emerg Infect Dis. 2011;17:584–591
4. Melegaro A, Choi Y, Pebody R, et al. Pneumococcal carriage in United Kingdom families: estimating serotype-specific transmission parameters from longitudinal data. Am J Epidemiol. 2007;166:228–235
5. Lipsitch M, Abdullahi O, D’Amour A, et al. Estimating rates of carriage acquisition and clearance and competitive ability for pneumococcal serotypes in Kenya with a Markov transition model. Epidemiology. 2012;23:510–519
6. Bogaert D, De Groot R, Hermans PW.. Streptococcus pneumoniae colonisation: the key to pneumococcal disease. Lancet Infect Dis. 2004;4:144–154
7. Pavia M, Bianco A, Nobile CG, et al. Efficacy of pneumococcal vaccination in children younger than 24 months: a meta-analysis. Pediatrics. 2009;123:e1103–e1110
8. Lucero MG, Dulalia VE, Parreno RN, et al. Pneumococcal conjugate vaccines for preventing vaccine-type invasive pneumococcal disease and pneumonia with consolidation on x-ray in children under two years of age. Cochrane Database Syst Rev. 2004:CD004977
9. Brueggemann AB, Pai R, Crook DW, et al. Vaccine escape recombinants emerge after pneumococcal vaccination in the United States. PLoS Pathog. 2007;3:e168
10. Nurhonen M, Auranen K.. Optimal serotype compositions for Pneumococcal conjugate vaccination under serotype replacement. PLoS Comput Biol. 2014;10:e1003477
11. van Hoek AJ, Sheppard CL, Andrews NJ, et al. Pneumococcal carriage in children and adults two years after introduction of the thirteen valent pneumococcal conjugate vaccine in England. Vaccine. 2014;32:4349–4355
12. Yildirim I, Hanage WP, Lipsitch M, et al. Serotype specific invasive capacity and persistent reduction in invasive pneumococcal disease. Vaccine. 2010;29:283–288
13. Miller E, Andrews NJ, Waight PA, et al. Herd immunity and serotype replacement 4 years after seven-valent pneumococcal conjugate vaccination in England and Wales: an observational cohort study. Lancet Infect Dis. 2011;11:760–768
14. Weinberger DM, Malley R, Lipsitch M.. Serotype replacement in disease following pneumococcal vaccination: a discussion of the evidence. Lancet. 2011;378:1962–1973
15. Hicks LA, Harrison LH, Flannery B, et al. Incidence of pneumococcal disease due to non-pneumococcal conjugate vaccine (PCV7) serotypes in the United States during the era of widespread PCV7 vaccination, 1998–2004. J Infect Dis. 2007;196:1346–1354
16. Vestrheim DF, Høiby EA, Bergsaker MR, et al. Indirect effect of conjugate pneumococcal vaccination in a 2+1 dose schedule. Vaccine. 2010;28:2214–2221
17. Lexau CA, Lynfield R, Danila R, et al.Active Bacterial Core Surveillance Team. Changing epidemiology of invasive pneumococcal disease among older adults in the era of pediatric pneumococcal conjugate vaccine. JAMA. 2005;294:2043–2051
18. Hanage WP, Finkelstein JA, Huang SS, et al. Evidence that pneumococcal serotype replacement in Massachusetts following conjugate vaccination is now complete. Epidemics. 2010;2:80–84
19. Steens A, Bergsaker MA, Aaberge IS, et al. Prompt effect of replacing the 7-valent pneumococcal conjugate vaccine with the 13-valent vaccine on the epidemiology of invasive pneumococcal disease in Norway. Vaccine. 2013;31:6232–6238
20. Kaplan SL, Barson WJ, Lin PL, et al. Early trends for invasive pneumococcal infections in children after the introduction of the 13-valent pneumococcal conjugate vaccine. Pediatr Infect Dis J. 2013;32:203–207
21. Guevara M, Ezpeleta C, Gil-Setas A, et al.Working Group for Surveillance of the Pneumococcal Disease in Navarre. Reduced incidence of invasive pneumococcal disease after introduction of the 13-valent conjugate vaccine in Navarre, Spain, 2001–2013. Vaccine. 2014;32:2553–2562
22. Moore CE, Paul J, Foster D, et al. Reduction of invasive pneumococcal disease three years after the introduction of the 13 valent conjugate vaccine in the Oxfordshire region, England. J Infect Dis. 2014
23. Vestrheim DF, Høiby EA, Aaberge IS, et al. Phenotypic and genotypic characterization of Streptococcus pneumoniae strains colonizing children attending day-care centers in Norway. J Clin Microbiol. 2008;46:2508–2518
24. Vestrheim DF, Høiby EA, Aaberge IS, et al. Impact of a pneumococcal conjugate vaccination program on carriage among children in Norway. Clin Vaccine Immunol. 2010;17:325–334
25. Child-care, 2010. [Statistics Norway Web Site]. Available at: http://www.ssb.no/en/utdanning/statistikker/kontantstotte/hvert-2-aar. Accessed May 2, 2011.
26. Riise O. Delay in the Norwegian Immunisation Programme [abstract]. 2014 Philadelphia IDWEEK
27. Clinical Breakpoints. [European Committee on Antimicrobial Susceptibility Testing (EUCAST) Web Site]. Available at: http://www.eucast.org/clinical_breakpoints/. Accessed November 20, 2013.
28. Enright MC, Spratt BG.. A multilocus sequence typing scheme for Streptococcus pneumoniae: identification of clones associated with serious invasive disease. Microbiology. 1998;144:3049–3060
29. Simpson EH.. Measurement of diversity. Nature. 1949;163:688
30. Dagan R, Patterson S, Juergens C, et al. Comparative immunogenicity and efficacy of 13-valent and 7-valent pneumococcal conjugate vaccines in reducing nasopharyngeal colonization: a randomized double-blind trial. Clin Infect Dis. 2013;57:952–962
31. Lee GM, Kleinman K, Pelton SI, et al. Impact of 13-valent pneumococcal conjugate vaccination on streptococcus pneumoniae carriage in young children in Massachusetts. J Pediatric Infect Dis Soc. 2014;3:23–32
32. Gounder PP, Bruce MG, Bruden DJ, et al. Effect of the 13-valent pneumococcal conjugate vaccine on nasopharyngeal colonization by Streptococcus pneumoniae–Alaska, 2008–2012. J Infect Dis. 2014;209:1251–1258
33. Ricketson LJ, Wood ML, Vanderkooi OG, et al.Calgary Streptococcus pneumoniae Epidemiology Research (CASPER) investigators. Trends in asymptomatic nasopharyngeal colonization with streptococcus pneumoniae after introduction of the 13-valent pneumococcal conjugate vaccine in Calgary, Canada. Pediatr Infect Dis J. 2014;33:724–730
34. Leino T, Hoti F, Syrjänen R, et al. Clustering of serotypes in a longitudinal study of Streptococcus pneumoniae carriage in three day care centres. BMC Infect Dis. 2008;8:173
35. Sá-Leão R, Nunes S, Brito-Avô A, et al. High rates of transmission of and colonization by Streptococcus pneumoniae and Haemophilus influenzae within a day care center revealed in a longitudinal study. J Clin Microbiol. 2008;46:225–234
36. Sleeman KL, Griffiths D, Shackley F, et al. Capsular serotype-specific attack rates and duration of carriage of Streptococcus pneumoniae in a population of children. J Infect Dis. 2006;194:682–688
37. Hammitt LL, Bruden DL, Butler JC, et al. Indirect effect of conjugate vaccine on adult carriage of Streptococcus pneumoniae: an explanation of trends in invasive pneumococcal disease. J Infect Dis. 2006;193:1487–1494
38. Loughlin AM, Hsu K, Silverio AL, et al. Direct and indirect effects of PCV13 on nasopharyngeal carriage of PCV13 unique pneumococcal serotypes in Massachusetts’ children. Pediatr Infect Dis J. 2014;33:504–510
39. Scott JR, Hanage WP, Lipsitch M, et al. Pneumococcal sequence type replacement among American Indian children: a comparison of pre- and routine-PCV7 eras. Vaccine. 2012;30:2376–2381
40. Hanage WP, Huang SS, Lipsitch M, et al. Diversity and antibiotic resistance among nonvaccine serotypes of Streptococcus pneumoniae carriage isolates in the post-heptavalent conjugate vaccine era. J Infect Dis. 2007;195:347–352
41. Vestrheim DF, Steinbakk M, Aaberge IS, et al. Postvaccination increase in serotype 19A pneumococcal disease in Norway is driven by expansion of penicillin-susceptible strains of the ST199 complex. Clin Vaccine Immunol. 2012;19:443–445
42. Kim DS, Shin SH, Lee HJ, et al. Immunogenicity and safety of 13-valent pneumococcal conjugate vaccine given to Korean children receiving routine pediatric vaccines. Pediatr Infect Dis J. 2013;32:266–273
43. Weckx LY, Thompson A, Berezin EN, et al.012 Study Group. A phase 3, randomized, double-blind trial comparing the safety and immunogenicity of the 7-valent and 13-valent pneumococcal conjugate vaccines, given with routine pediatric vaccinations, in healthy infants in Brazil. Vaccine. 2012;30:7566–7572
44. Kieninger DM, Kueper K, Steul K, et al.006 study group. Safety, tolerability, and immunologic noninferiority of a 13-valent pneumococcal conjugate vaccine compared to a 7-valent pneumococcal conjugate vaccine given with routine pediatric vaccinations in Germany. Vaccine. 2010;28:4192–4203
45. Scott DA, Komjathy SF, Hu BT, et al. Phase 1 trial of a 13-valent pneumococcal conjugate vaccine in healthy adults. Vaccine. 2007;25:6164–6166
46. Nunes MC, Madhi SA.. Review on the immunogenicity and safety of PCV-13 in infants and toddlers. Expert Rev Vaccines. 2011;10:951–980
47. Miller E, Andrews NJ, Waight PA, et al. Effectiveness of the new serotypes in the 13-valent pneumococcal conjugate vaccine. Vaccine. 2011;29:9127–9131
48. . Statistics from the Norwegian Prescription Database. [Norwegian Institute for Public Health Web Site]. Available at: http://www.norpd.no/Prevalens.aspx. Accessed July 8, 2014
Keywords:

Streptococcus pneumoniae; pneumococcal carriage; pneumococcal conjugate vaccine; serotype diversity; multilocus sequence typing; antibiotic resistance

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