Related Experiment Video
Updated: May 7, 2026

Capsular Serotyping of Streptococcus pneumoniae Using the Quellung Reaction
Published on: February 24, 2014
Serotype epidemiology and antibiotic resistance of pneumococcal isolates colonizing infants in Botswana (2016-2019)
Jillian H Hurst1, Yazdani B Shaik-Dasthagirisaheb2, Loc Truong2
1Division of Pediatric Infectious Diseases, Duke School of Medicine, Durham, North Carolina, United States of America.
Insights
Pneumococcal conjugate vaccine (PCV-13) use in Botswana showed vaccine serotypes colonizing infants, alongside rising penicillin resistance. Amoxicillin remains effective for non-meningeal infections, while cephalosporins are recommended for meningitis.
Area of Science:
- Pediatrics
- Infectious Diseases
- Vaccinology
Background:
- Botswana implemented the 13-valent pneumococcal conjugate vaccine (PCV-13) in 2012.
- High PCV-13 coverage (over 90%) was achieved by 2014.
- Concerns exist regarding potential increases in non-vaccine serotypes and antibiotic resistance post-PCV introduction.
Purpose of the Study:
- To characterize pneumococcal serotype epidemiology in infants.
- To assess antibiotic resistance patterns in pneumococcal isolates.
- To evaluate the impact of PCV-13 on serotype distribution and resistance in Botswana.
Main Methods:
- Nasopharyngeal samples were collected from infants (≤12 months) in southeastern Botswana (2016-2019).
- Pneumococcal isolates were identified and serotyped using the Quellung reaction.
- Antibiotic susceptibility was determined using E-tests for minimum inhibitory concentrations.
Main Results:
- 264 pneumococcal isolates were cultured from 150 infants; 81% had received at least one PCV-13 dose.
- PCV-13 serotypes comprised 27% of isolates, with 19F, 19A, and 6A being most common.
- Non-vaccine serotypes 23B, 21, and 16F were most frequent; penicillin resistance increased significantly (41% to 71%), while amoxicillin resistance remained low (1%).
Conclusions:
- PCV-13 serotypes were prevalent in infant nasopharyngeal carriage four years post-introduction.
- Low amoxicillin resistance supports its use for non-meningeal pneumococcal infections.
- Rising penicillin resistance and low ceftriaxone resistance support cephalosporins for empirical meningitis treatment.
Background:
In 2012, Botswana introduced 13-valent pneumococcal conjugate vaccine (PCV-13) to its childhood immunization program in a 3+0 schedule, achieving coverage rates of above 90% by 2014. In other settings, PCV introduction has been followed by an increase in carriage or disease caused by non-vaccine serotypes, including some serotypes with a high prevalence of antibiotic resistance.
Methods:
We characterized the serotype epidemiology and antibiotic resistance of pneumococcal isolates cultured from nasopharyngeal samples collected from infants (≤12 months) in southeastern Botswana between 2016 and 2019. Capsular serotyping was performed using the Quellung reaction. E-tests were used to determine minimum inhibitory concentrations for common antibiotics.
Results:
We cultured 264 pneumococcal isolates from samples collected from 150 infants. At the time of sample collection, 81% of infants had received at least one dose of PCV-13 and 53% had completed the three-dose series. PCV-13 serotypes accounted for 27% of isolates, with the most prevalent vaccine serotypes being 19F (n = 20, 8%), 19A (n = 16, 6%), and 6A (n = 10, 4%). The most frequently identified non-vaccine serotypes were 23B (n = 29, 11%), 21 (n = 12, 5%), and 16F (n = 11, 4%). Only three (1%) pneumococcal isolates were resistant to amoxicillin; however, we observed an increasing prevalence of penicillin resistance using the meningitis breakpoint (2016: 41%, 2019: 71%; Cochran-Armitage test for trend, p = 0.0003) and non-susceptibility to trimethoprim-sulfamethoxazole (2016: 55%, 2019: 79%; p = 0.04). Three (1%) isolates were multi-drug resistant.
Conclusions:
PCV-13 serotypes accounted for a substantial proportion of isolates colonizing infants in Botswana during a four-year period starting four years after vaccine introduction. A low prevalence of amoxicillin resistance supports its continued use as the first-line agent for non-meningeal pneumococcal infections. The observed increase in penicillin resistance at the meningitis breakpoint and the low prevalence of resistance to ceftriaxone supports use of third-generation cephalosporins for empirical treatment of suspected bacterial meningitis.
Related Concept Videos
Development of Antibiotic Resistance
Atypical Pneumonia
Mechanism of Antibiotic Resistance in MRSA
Clinical Significance of Antibiotic Resistance
Bacterial Meningitis II: Pathophysiology

