PLoS One. 2016 Mar 17;11(3):e0151428. doi: 10.1371/journal.pone.0151428. eCollection 2016.

The Relevance of a Novel Quantitative Assay to Detect up to 40 Major Streptococcus pneumoniaeSerotypes Directly in Clinical Nasopharyngeal and Blood Specimens.

Messaoudi M1, Milenkov M1, Albrich WC2,3, van der Linden MP4, Bénet T5, Chou M6, Sylla M7, Barreto Costa P8, Richard N9, Klugman KP3,10,Endtz HP1,11, Paranhos-Baccalà G1, Telles JN1.

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Abstract

For epidemiological and surveillance purposes, it is relevant to monitor the distribution and dynamics of Streptococcus pneumoniaeserotypes. Conventional serotyping methods do not provide rapid or quantitative information on serotype loads. Quantitative serotyping may enable prediction of the invasiveness of a specific serotype compared to other serotypes carried. Here, we describe a novel, rapid multiplex real-time PCR assay for identification and quantification of the 40 most prevalent pneumococcal serotypes and the assay impacts in pneumonia specimens from emerging and developing countries. Eleven multiplex PCR to detect 40 serotypes or serogroups were optimized. Quantification was enabled by reference to standard dilutions of known bacterial load. Performance of the assay was evaluated to specifically type and quantify S. pneumoniae in nasopharyngeal and blood samples from adult and pediatric patients hospitalized with pneumonia (n = 664) from five different countries. Serogroup 6 was widely represented in nasopharyngeal specimens from all five cohorts. The most frequent serotypes in the French, South African, and Brazilian cohorts were 1 and 7A/F, 3 and 19F, and 14, respectively. When both samples were available, the serotype in blood was always present as carriage with other serotypes in the nasopharynx. Moreover, the ability of a serotype to invade the bloodstream may be linked to its nasopharyngeal load. The mean nasopharyngeal concentration of the serotypes that moved to the blood was 3 log-fold higher than the ones only found in the nasopharynx. This novel, rapid, quantitative assay may potentially predict some of the S. pneumoniae serotypes invasiveness and assessment of pneumococcal serotype distribution.

PMID: 26986831 [PubMed - in process]