Evaluation of pneumococcal serotyping of nasopharyngeal-carriage isolates by latex agglutination, whole-genome sequencing (PneumoCaT), and DNA microarray in a high-pneumococcal-carriage-prevalence population in Malawi

Todd D. Swarthout, Andrea Gori, Naor Bar-Zeev, Arox W. Kamng'Ona, Thandie S. Mwalukomo, Farouck Bonomali, Roseline Nyirenda, Comfort Brown, Jacquline Msefula, Dean Everett, Charles Mwansambo, Katherine Gould, Jason Hinds, Robert S. Heyderman, Neil French

Research output: Contribution to journalArticlepeer-review

Abstract

Accurate assessment of the serotype distribution associated with pneumococcal colonization and disease is essential for evaluating and formulating pneumococcal vaccines and for informing vaccine policy. For this reason, we evaluated the concordance between pneumococcal serotyping results by latex agglutination, whole-genome sequencing (WGS) with PneumoCaT, and DNA microarray for samples from community carriage surveillance in Blantyre, Malawi. Nasopharyngeal swabs were collected according to WHO recommendations between 2015 and 2017 by using stratified random sampling among study populations. Participants included healthy children 3 to 6 years old (vaccinated with the 13-valent pneumococcal conjugate vaccine [PCV13] as part of the Expanded Program on Immunization [EPI]), healthy children 5 to 10 years old (age-ineligible for PCV13), and HIV-infected adults (18 to 40 years old) on antiretroviral therapy (ART). For phenotypic serotyping, we used a 13-valent latex kit (Statens Serum Institut [SSI], Denmark). For genomic serotyping, we applied the PneumoCaT pipeline to whole-genome sequence libraries. For molecular serotyping by microarray, we used the BUGS Bioscience Senti-SP microarray. A total of 1,347 samples were analyzed. Concordance was 90.7% (95% confidence interval [CI], 89.0 to 92.2%) between latex agglutination and PneumoCaT, 95.2% (95% CI, 93.9 to 96.3%) between latex agglutination and the microarray, and 96.6% (95% CI, 95.5 to 97.5%) between the microarray and PneumoCaT. By detecting additional vaccine serotype (VT) pneumococci carried at low relative abundances (median, 8%), the microarray increased VT detection by 31.5% over that by latex serotyping. To conclude, all three serotyping methods were highly concordant in identifying dominant serotypes. Latex serotyping is accurate in identifying vaccine serotypes and requires the least expertise and resources for field implementation and analysis. However, WGS, which adds population structure, and microarray, which adds multiple-serotype carriage, should be considered at regional reference laboratories for investigating the importance of vaccine serotypes at low relative abundances in transmission and disease.

Original languageEnglish (US)
Article numbere02103-20
JournalJournal of clinical microbiology
Volume59
Issue number1
DOIs
StatePublished - Jan 2021

Keywords

  • Africa
  • Latex agglutination
  • Methodology
  • Microarray
  • Serotyping
  • Streptococcus pneumoniae
  • Whole-genome sequencing

ASJC Scopus subject areas

  • Microbiology (medical)

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