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Abstract

Despite the introduction of pneumococcal conjugate vaccines (PCVs), still remains an important cause of morbidity and mortality, especially among children under 5 years in sub-Saharan Africa. We sought to determine the distribution of serotypes, lineages and antimicrobial resistance of from carriage and disease among children presenting to health facilities, 5–6 years after the introduction of PCV10 in Ethiopia.

Whole-genome sequencing (WGS) was performed on 103  (86 from nasopharyngeal swabs, 4 from blood and 13 from middle ear discharge) isolated from children aged <15 years at 3 healthcare facilities in Addis Ababa, Ethiopia, from September 2016 to August 2017. Using the WGS data, serotypes were predicted, isolates were assigned to clonal complexes, global pneumococcal sequence clusters (GPSCs) were inferred and screening for alleles and mutations that confer resistance to antibiotics was performed using multiple bioinformatic pipelines.

The 103 . isolates were assigned to 38 serotypes (including nontypeable) and 46 different GPSCs. The most common serotype was serotype 19A. Common GPSCs were GPSC1 [14.6% (15/103), sequence type (ST) 320, serotype 19A], GPSC268 [8.7% (9/103), ST 6882 and novel STs; serotypes 16F, 11A and 35A] and GPSC10 [8.7% (9/103), STs 2013, 230 and 8804; serotype 19A]. The four invasive isolates were serotype 19A (=2) and serotype 33C (=2). Resistance to penicillin (>0.06 µg ml, CLSI meningitis cutoff) was predicted in 57% (59/103) of the isolates, and 43% (25/58) penicillin-binding protein allele combinations were predicted to be associated with penicillin resistance. Resistance mutations in () and/or (indel between fifty-sixth and sixty-seventh ) were identified among 66% (68/103) of the isolates, whilst tetracycline () and macrolide ( and ) resistance genes were found in 46.6% (48/103), 20.4% (21/103) and 20.4% (21/103) of the isolates, respectively. Multidrug resistance (MDR) (≥3 antibiotic classes) was observed in 31.1% (32/103) of the isolates. GPSC1 and GPSC10 accounted for 46.8% (15/32) and 18.7% (6/32) of the overall MDR.

Five to 6 years after the introduction of PCV10 in Ethiopia, the obtained from carriage and disease among paediatric patients showed diverse serotype and pneumococcal lineages. The most common serotype identified was 19A, expressed by the MDR lineages GPSC1 and GPSC10, which is not covered by PCV10 but is included in PCV13. Continued assessment of the impact of PCV on the population structure of in Ethiopia is warranted during and after PCV13 introduction.

Funding
This study was supported by the:
  • Wellcome Sanger Institute (Award 098051 and 206194)
    • Principal Award Recipient: StephenD Bentley
  • Bill and Melinda Gates Foundation (Award OPP1034556)
    • Principal Award Recipient: StephenD Bentley
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2025-03-18
2026-02-16

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References

  1. Troeger C, Blacker B, Khalil IA, Rao PC, Cao J et al. Estimates of the global, regional, and national morbidity, mortality, and aetiologies of lower respiratory infections in 195 countries, 1990–2016: a systematic analysis for the global burden of disease study 2016. Lancet Infect Dis 2018; 18:1191–1210 [View Article]
    [Google Scholar]
  2. Wahl B, O’Brien KL, Greenbaum A, Majumder A, Liu L et al. Burden of Streptococcus pneumoniae and Haemophilus influenzae type b disease in children in the era of conjugate vaccines: global, regional, and national estimates for 2000-15. Lancet Glob Health 2018; 6:e744–e757 [View Article] [PubMed]
    [Google Scholar]
  3. Musher DM, Anderson R, Feldman C. The remarkable history of pneumococcal vaccination: an ongoing challenge. Pneumonia 2022; 14:5 [View Article] [PubMed]
    [Google Scholar]
  4. Fleming-Dutra KE, Conklin L, Loo JD, Knoll MD, Park DE et al. Systematic review of the effect of pneumococcal conjugate vaccine dosing schedules on vaccine-type nasopharyngeal carriage. Pediatr Infect Dis J 2014; 33 Suppl 2:S152–60 [View Article] [PubMed]
    [Google Scholar]
  5. Roca A, Bojang A, Bottomley C, Gladstone RA, Adetifa JU et al. Effect on nasopharyngeal pneumococcal carriage of replacing PCV7 with PCV13 in the expanded programme of immunization in the Gambia. Vaccine 2015; 33:7144–7151 [View Article] [PubMed]
    [Google Scholar]
  6. Adamu AL, Ojal J, Abubakar IA, Odeyemi KA, Bello MM et al. The impact of introduction of the 10-valent pneumococcal conjugate vaccine on pneumococcal carriage in Nigeria. Nat Commun 2023; 14:2666 [View Article] [PubMed]
    [Google Scholar]
  7. Conklin L, Loo JD, Kirk J, Fleming-Dutra KE, Knoll MD et al. Systematic review of the effect of pneumococcal conjugate vaccine dosing schedules on vaccine-type invasive pneumococcal disease among young children. Pediatr Infect Dis J 2014; 33 Suppl 2:S109–18 [View Article] [PubMed]
    [Google Scholar]
  8. von Gottberg A, Kleynhans J, de Gouveia L, Tempia S, Meiring S et al. Long-term effect of pneumococcal conjugate vaccines on invasive pneumococcal disease incidence among people of all ages from national, active, laboratory-based surveillance in South Africa, 2005–19: a cohort observational study. Lancet Glob Health 2024; 12:e1470–e1484 [View Article]
    [Google Scholar]
  9. Mackenzie GA, Hill PC, Jeffries DJ, Ndiaye M, Sahito SM et al. Impact of the introduction of pneumococcal conjugate vaccination on invasive pneumococcal disease and pneumonia in the Gambia: 10 years of population-based surveillance. Lancet Infect Dis 2021; 21:1293–1302 [View Article] [PubMed]
    [Google Scholar]
  10. Loo JD, Conklin L, Fleming-Dutra KE, Knoll MD, Park DE et al. Systematic review of the indirect effect of pneumococcal conjugate vaccine dosing schedules on pneumococcal disease and colonization. Pediatr Infect Dis J 2014; 33 Suppl 2:S161–71 [View Article] [PubMed]
    [Google Scholar]
  11. Chan J, Nguyen CD, Dunne EM, Kim Mulholland E, Mungun T et al. Using pneumococcal carriage studies to monitor vaccine impact in low- and middle-income countries. Vaccine 2019; 37:6299–6309 [View Article] [PubMed]
    [Google Scholar]
  12. Weinberger DM, Malley R, Lipsitch M. Serotype replacement in disease after pneumococcal vaccination. The Lancet 2011; 378:1962–1973 [View Article]
    [Google Scholar]
  13. Hanquet G, Krizova P, Dalby T, Ladhani SN, Nuorti JP et al. Serotype replacement after introduction of 10-valent and 13-valent pneumococcal conjugate vaccines in 10 countries, Europe. Emerg Infect Dis 2022; 28:137–138 [View Article] [PubMed]
    [Google Scholar]
  14. Ngocho JS, Magoma B, Olomi GA, Mahande MJ, Msuya SE et al. Effectiveness of pneumococcal conjugate vaccines against invasive pneumococcal disease among children under five years of age in Africa: a systematic review. PLoS One 2019; 14:e0212295 [View Article] [PubMed]
    [Google Scholar]
  15. Gladstone RA, Lo SW, Goater R, Yeats C, Taylor B et al. Visualizing variation within Global Pneumococcal Sequence Clusters (GPSCS) and country population snapshots to contextualize pneumococcal isolates. Microb Genom 2020; 6:1–13 [View Article] [PubMed]
    [Google Scholar]
  16. Gladstone RA, Lo SW, Lees JA, Croucher NJ, van Tonder AJ et al. International genomic definition of pneumococcal lineages, to contextualise disease, antibiotic resistance and vaccine impact. EBioMedicine 2019; 43:338–346 [View Article] [PubMed]
    [Google Scholar]
  17. Hicks LA, Harrison LH, Flannery B, Hadler JL, Schaffner W 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 [View Article] [PubMed]
    [Google Scholar]
  18. Balsells E, Dagan R, Yildirim I, Gounder PP, Steens A et al. The relative invasive disease potential of Streptococcus pneumoniae among children after PCV introduction: a systematic review and meta-analysis. J Infect 2018; 77:368–378 [View Article] [PubMed]
    [Google Scholar]
  19. Brandileone M-C de C, Zanella RC, Almeida SCG, Cassiolato AP, Lemos APS de et al. Long-term effect of 10-valent pneumococcal conjugate vaccine on nasopharyngeal carriage of Streptococcus pneumoniae in children in Brazil. Vaccine 2019; 37:5357–5363 [View Article] [PubMed]
    [Google Scholar]
  20. Cassiolato AP, Almeida SCG, Andrade AL, Minamisava R, Cristina de Cunto Brandileone M. Expansion of the multidrug-resistant clonal complex 320 among invasive Streptococcus pneumoniae serotype 19A after the introduction of a ten-valent pneumococcal conjugate vaccine in Brazil. PLoS One 2018; 13:e0208211 [View Article] [PubMed]
    [Google Scholar]
  21. Isturiz R, Sings HL, Hilton B, Arguedas A, Reinert RR et al. Streptococcus pneumoniae serotype 19A: worldwide epidemiology. Expert Rev Vaccines 2017; 16:1007–1027 [View Article] [PubMed]
    [Google Scholar]
  22. Corcoran M, Mereckiene J, Cotter S, Murchan S, Lo SW et al. Using genomics to examine the persistence of Streptococcus pneumoniae serotype 19A in Ireland and the emergence of a sub-clade associated with vaccine failures. Vaccine 2021; 39:5064–5073 [View Article] [PubMed]
    [Google Scholar]
  23. Ethiopia M of HFR of Introducing pneumococcal conjugate vaccine in ethiopia: training manual for health workers; 2011
  24. Negash AA, Asrat D, Abebe W, Hailemariam T, Gebre M et al. Pneumococcal carriage, serotype distribution, and risk factors in children with community-acquired pneumonia, 5 years after introduction of the 10-valent pneumococcal conjugate vaccine in Ethiopia. Open Forum Infect Dis 2019; 6:ofz259 [View Article] [PubMed]
    [Google Scholar]
  25. Negash AA, Asrat D, Abebe W, Hailemariam T, Gebre M et al. Pneumococcal serotype 19A is the major cause of pediatric acute otitis media with ruptured tympanic membrane in Addis Ababa, Ethiopia, 5 years after the introduction of the ten-valent pneumococcal conjugate vaccine. Int J Pediatr Otorhinolaryngol 2019; 126:109638 [View Article] [PubMed]
    [Google Scholar]
  26. Negash AA, Asrat D, Abebe W, Aseffa A, Vaneechoutte M. Phenotypic and molecular characterization of penicillin and macrolide-resistant Streptococcus pneumoniae serotypes among pediatric patients in Addis Ababa, Ethiopia. Infect Drug Resist 2021; 14:1765–1772 [View Article] [PubMed]
    [Google Scholar]
  27. Negash AA, Asrat D, Abebe W, Hailemariam T, Wubete A et al. Etiology, antibiotic susceptibility and prognostic factors of pediatric community-acquired sepsis in Addis Ababa, Ethiopia. J Infect Dev Ctries 2021; 15:113–122 [View Article] [PubMed]
    [Google Scholar]
  28. Sørensen UBS. Typing of pneumococci by using 12 pooled antisera. J Clin Microbiol 1993; 31:2097–2100 [View Article]
    [Google Scholar]
  29. Page AJ, De Silva N, Hunt M, Quail MA, Parkhill J et al. Robust high-throughput prokaryote de novo assembly and improvement pipeline for Illumina data. Microb Genom 2016; 2:e000083 [View Article] [PubMed]
    [Google Scholar]
  30. Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics 2014; 30:2068–2069 [View Article] [PubMed]
    [Google Scholar]
  31. Epping L, van Tonder AJ, Gladstone RA, Bentley SD, Page AJ et al. SeroBA: rapid high-throughput serotyping of Streptococcus pneumoniae from whole genome sequence data. Microb Genom 2018; 4:1–6 [View Article] [PubMed]
    [Google Scholar]
  32. J. Page A, Taylor B, A. Keane J. Multilocus sequence typing by blast from de novo assemblies against PubMLST. J Open Source Softw 2016; 1:118 [View Article]
    [Google Scholar]
  33. Lees JA, Harris SR, Tonkin-Hill G, Gladstone RA, Lo SW et al. Fast and flexible bacterial genomic epidemiology with PopPUNK. Genome Res 2019; 29:304–316 [View Article]
    [Google Scholar]
  34. Metcalf BJ, Gertz RE Jr, Gladstone RA, Walker H, Sherwood LK et al. Strain features and distributions in pneumococci from children with invasive disease before and after 13-valent conjugate vaccine implementation in the USA. Clin Microbiol Infect 2016; 22:60 [View Article] [PubMed]
    [Google Scholar]
  35. Metcalf BJ, Chochua S, Gertz RE Jr, Li Z, Walker H et al. Using whole genome sequencing to identify resistance determinants and predict antimicrobial resistance phenotypes for year 2015 invasive pneumococcal disease isolates recovered in the United States. Clin Microbiol Infect 2016; 22:1002 [View Article] [PubMed]
    [Google Scholar]
  36. Price MN, Dehal PS, Arkin AP. FastTree 2--approximately maximum-likelihood trees for large alignments. PLoS One 2010; 5:e9490 [View Article] [PubMed]
    [Google Scholar]
  37. Sigaúque B, Moiane B, Massora S, Pimenta F, Verani JR et al. Early declines in vaccine-type pneumococcal carriage in children less than 5 years old after introduction of 10-valent pneumococcal conjugate vaccine in Mozambique. Pediatr Infect Dis J 2018; 37:1054–1060 [View Article]
    [Google Scholar]
  38. Shrestha S, Gurung M, Amatya P, Bijukchhe S, Bose AS et al. Effect of the of 10-valent pneumococcal conjugate vaccine in Nepal 4 years after introduction: an observational cohort study. Lancet Glob Health 2022; 10:e1494–e1504 [View Article] [PubMed]
    [Google Scholar]
  39. Nisar MI, Jehan F, Shahid S, Ahmed S, Shakoor S et al. Serotype-specific effectiveness against pneumococcal carriage and serotype replacement after ten-valent Pneumococcal Conjugate Vaccine (PCV10) introduction in Pakistan. PLoS One 2022; 17:e0262466 [View Article] [PubMed]
    [Google Scholar]
  40. Ekinci E, Van Heirstraeten L, Willen L, Desmet S, Wouters I et al. Serotype 19A and 6C account for one-third of pneumococcal carriage among Belgian day-care children four years after a shift to a lower-valent PCV. J Pediatric Infect Dis Soc 2023; 12:36–42 [View Article] [PubMed]
    [Google Scholar]
  41. Desmet S, Theeten H, Laenen L, Cuypers L, Maes P et al. Characterization of emerging serotype 19A pneumococcal strains in invasive disease and carriage, Belgium. Emerg Infect Dis 2022; 28:1606–1614 [View Article] [PubMed]
    [Google Scholar]
  42. Macaj M, Perdochova L, Jakubikova J. Streptococcus pneumoniae as cause of acute otitis media (AOM) in Slovak children in the pneumococcal conjugate vaccine era (2008-2019). Vaccine 2023; 41:452–459 [View Article] [PubMed]
    [Google Scholar]
  43. Jarovsky D, Berezin EN. Impact of PCV10 on pediatric pneumococcal disease burden in Brazil: time for new recommendations?. J Pediatr 2023; 99:S46–S56 [View Article]
    [Google Scholar]
  44. Almeida SCG, Lo SW, Hawkins PA, Gladstone RA, Cassiolato AP et al. Genomic surveillance of invasive Streptococcus pneumoniae isolates in the period pre-PCV10 and post-PCV10 introduction in Brazil. Microb Genom 2021; 7:1–13 [View Article] [PubMed]
    [Google Scholar]
  45. Egorova E, Kumar N, Gladstone RA, Urban Y, Voropaeva E et al. Key features of pneumococcal isolates recovered in Central and Northwestern Russia in 2011-2018 determined through whole-genome sequencing. Microb Genom 2022; 8:mgen000851 [View Article] [PubMed]
    [Google Scholar]
  46. Tonkin-Hill G, Ling C, Chaguza C, Salter SJ, Hinfonthong P et al. Pneumococcal within-host diversity during colonization, transmission and treatment. Nat Microbiol 2022; 7:1791–1804 [View Article] [PubMed]
    [Google Scholar]
  47. Balsells E, Guillot L, Nair H, Kyaw MH. Serotype distribution of Streptococcus pneumoniae causing invasive disease in children in the post-PCV era: a systematic review and meta-analysis. PLoS One 2017; 12:e0177113 [View Article] [PubMed]
    [Google Scholar]
  48. Croucher NJ, Harris SR, Fraser C, Quail MA, Burton J et al. Rapid pneumococcal evolution in response to clinical interventions. Science 2011; 331:430–434 [View Article] [PubMed]
    [Google Scholar]
  49. Simões AS, Pereira L, Nunes S, Brito-Avô A, De Lencastre H et al. Clonal evolution leading to maintenance of antibiotic resistance rates among colonizing Pneumococci in the PCV7 era in Portugal. J Clin Microbiol 2011; 49:2810–2817 [View Article] [PubMed]
    [Google Scholar]
  50. Gherardi G, D’Ambrosio F, Visaggio D, Dicuonzo G, Del Grosso M et al. Erratum: serotype and clonal evolution of penicillin-nonsusceptible invasive Streptococcus pneumoniae in the 7-valent pneumococcal conjugate vaccine era in Italy. Antimicrob Agents Chemother 2012; 56:4965–4968 [View Article] [PubMed]
    [Google Scholar]
  51. Ndlangisa KM, du Plessis M, Lo S, de Gouveia L, Chaguza C et al. A Streptococcus pneumoniae lineage usually associated with pneumococcal conjugate vaccine (PCV) serotypes is the most common cause of serotype 35B invasive disease in South Africa, following routine use of PCV. Microb Genom 2022; 8:000746 [View Article] [PubMed]
    [Google Scholar]
  52. Hurmic O, Grall N, Al Nakib M, Poyart C, Grondin S et al. Evidence of a clonal expansion of Streptococcus pneumoniae serotype 19A in adults as in children assessed by the DiversiLab® system. Eur J Clin Microbiol Infect Dis 2014; 33:2067–2073 [View Article] [PubMed]
    [Google Scholar]
  53. Ardanuy C, Marimón JM, Calatayud L, Giménez M, Alonso M et al. Epidemiology of invasive pneumococcal disease in older people in Spain (2007-2009): implications for future vaccination strategies. PLoS One 2012; 7:e43619 [View Article] [PubMed]
    [Google Scholar]
  54. Mayanskiy N, Savinova T, Alyabieva N, Ponomarenko O, Brzhozovskaya E et al. Antimicrobial resistance, penicillin-binding protein sequences, and pilus islet carriage in relation to clonal evolution of Streptococcus pneumoniae serotype 19A in Russia, 2002-2013. Epidemiol Infect 2017; 145:1708–1719 [View Article] [PubMed]
    [Google Scholar]
  55. Lo SW, Mellor K, Cohen R, Alonso AR, Belman S et al. Emergence of a multidrug-resistant and virulent Streptococcus pneumoniae lineage mediates serotype replacement after PCV13: an international whole-genome sequencing study. Lancet Microbe 2022; 3:e735–e743 [View Article] [PubMed]
    [Google Scholar]
  56. Gagetti P, Lo SW, Hawkins PA, Gladstone RA, Regueira M et al. Population genetic structure, serotype distribution and antibiotic resistance of Streptococcus pneumoniae causing invasive disease in children in Argentina. Microb Genom 2021; 7:000636 [View Article] [PubMed]
    [Google Scholar]
  57. Lo SW, Gladstone RA, van Tonder AJ, Lees JA, du Plessis M et al. Pneumococcal lineages associated with serotype replacement and antibiotic resistance in childhood invasive pneumococcal disease in the post-PCV13 era: an international whole-genome sequencing study. Lancet Infect Dis 2019; 19:759–769 [View Article] [PubMed]
    [Google Scholar]
  58. Kalizang’oma A, Chaguza C, Gori A, Davison C, Beleza S et al. Streptococcus pneumoniae serotypes that frequently colonise the human nasopharynx are common recipients of penicillin-binding protein gene fragments from Streptococcus mitis. Microb Genom 2021; 7:000622 [View Article] [PubMed]
    [Google Scholar]
  59. Nisar MI, Shahid S, Jehan F, Ahmed S, Shakoor S et al. Antimicrobial resistance in pneumococcal carriage isolates from children under 2 years of age in rural Pakistan. Microbiol Spectr 2021; 9:e0101921 [View Article] [PubMed]
    [Google Scholar]
  60. Kobayashi M, Bigogo G, Kim L, Mogeni OD, Conklin LM et al. Impact of 10valent pneumococcal conjugate vaccine introduction on pneumococcal carriage and antibiotic susceptibility patterns among children aged <5 years and adults with human immunodeficiency virus infection: Kenya, 2009-2013. Clin Infect Dis 2020; 70:814–826 [View Article] [PubMed]
    [Google Scholar]
  61. Klugman KP, Black S. Impact of existing vaccines in reducing antibiotic resistance: primary and secondary effects. Proc Natl Acad Sci USA 2018; 115:12896–12901 [View Article] [PubMed]
    [Google Scholar]
  62. Moore MR, Gertz RE Jr, Woodbury RL, Barkocy-Gallagher GA, Schaffner W et al. Population snapshot of emergent Streptococcus pneumoniae serotype 19A in the United States, 2005. J Infect Dis 2008; 197:1016–1027 [View Article] [PubMed]
    [Google Scholar]
  63. Dagan R, Givon-Lavi N, Leibovitz E, Greenberg D, Porat N. Introduction and proliferation of multidrug-resistant Streptococcus pneumoniae serotype 19A clones that cause acute otitis media in an unvaccinated population. J Infect Dis 2009; 199:776–785 [View Article] [PubMed]
    [Google Scholar]
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