Revisiting pneumococcal carriage by use of broth enrichment and PCR techniques for enhanced detection of carriage and serotypes

Maria Da Gloria Carvalho, Fabiana C. Pimenta, Delois Jackson, Alexis Roundtree, Yusra Ahmad, Eugene V. Millar, Katherine L O'Brien, Cynthia G. Whitney, Adam L. Cohen, Bernard W. Beall

Research output: Contribution to journalArticle

Abstract

The measurement of pneumococcal carriage in the nasopharyngeal reservoir is subject to potential confounders that include low-density and multiple-strain colonization. To compare different methodologies, we picked a random sampling of 100 nasopharyngeal specimens recovered from infants less than 2 years of age who were previously assessed for pneumococcal carriage and serotypes by a conventional method that used direct plating from the transport/storage medium (50 specimens were culture negative and 50 specimens were culture positive for pneumococci). We used a broth enrichment approach and a conventional PCR approach (with and without broth enrichment) to determine pneumococcal carriage and serotypes, and the results were compared to the initial conventional culture-based results. Additionally, we used a lytA-targeted real-time PCR for pneumococcal detection. Broth enrichment for both the culture-based and the PCR-based methods enhanced the isolation of pneumococci and detection of serotype diversity, with the most effective serotype deduction method being one that used broth enrichment prior to sequential multiplex PCR. Similarly, we also found that broth enrichment followed by the lytA-specific real-time PCR was the most sensitive for the detection of apparent pneumococcal carriage. The broth enrichment, conventional multiplex PCR, and real-time PCR approaches used in this study were effective in detecting pneumococcal carriage in the 50 specimens that were negative by conventional direct plating from transport medium (range of numbers of positive specimens, 8/50 to 22/50 [16 to 44%]), and the three different serotyping approaches that used broth enrichment increased the number of serotype identifications from the 100 specimens (12 to 29 additional serotype identifications to be positive). A PCR-based approach that employed a broth enrichment step appeared to best enhance the detection of mixed serotypes and low-density pneumococcal carriage.

Original languageEnglish (US)
Pages (from-to)1611-1618
Number of pages8
JournalJournal of Clinical Microbiology
Volume48
Issue number5
DOIs
StatePublished - May 2010

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Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
Multiplex Polymerase Chain Reaction
Streptococcus pneumoniae
Serotyping
Serogroup

ASJC Scopus subject areas

  • Microbiology (medical)

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Carvalho, M. D. G., Pimenta, F. C., Jackson, D., Roundtree, A., Ahmad, Y., Millar, E. V., ... Beall, B. W. (2010). Revisiting pneumococcal carriage by use of broth enrichment and PCR techniques for enhanced detection of carriage and serotypes. Journal of Clinical Microbiology, 48(5), 1611-1618. https://doi.org/10.1128/JCM.02243-09

Revisiting pneumococcal carriage by use of broth enrichment and PCR techniques for enhanced detection of carriage and serotypes. / Carvalho, Maria Da Gloria; Pimenta, Fabiana C.; Jackson, Delois; Roundtree, Alexis; Ahmad, Yusra; Millar, Eugene V.; O'Brien, Katherine L; Whitney, Cynthia G.; Cohen, Adam L.; Beall, Bernard W.

In: Journal of Clinical Microbiology, Vol. 48, No. 5, 05.2010, p. 1611-1618.

Research output: Contribution to journalArticle

Carvalho, MDG, Pimenta, FC, Jackson, D, Roundtree, A, Ahmad, Y, Millar, EV, O'Brien, KL, Whitney, CG, Cohen, AL & Beall, BW 2010, 'Revisiting pneumococcal carriage by use of broth enrichment and PCR techniques for enhanced detection of carriage and serotypes', Journal of Clinical Microbiology, vol. 48, no. 5, pp. 1611-1618. https://doi.org/10.1128/JCM.02243-09
Carvalho, Maria Da Gloria ; Pimenta, Fabiana C. ; Jackson, Delois ; Roundtree, Alexis ; Ahmad, Yusra ; Millar, Eugene V. ; O'Brien, Katherine L ; Whitney, Cynthia G. ; Cohen, Adam L. ; Beall, Bernard W. / Revisiting pneumococcal carriage by use of broth enrichment and PCR techniques for enhanced detection of carriage and serotypes. In: Journal of Clinical Microbiology. 2010 ; Vol. 48, No. 5. pp. 1611-1618.
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