Density and duration of experimental human pneumococcal carriage

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Density and duration of experimental human pneumococcal carriage J.F. Gritzfeld, A.J.H. Cremers, G. Ferwerda, D.M. Ferreira, A. Kadioglu, P.W.M. Hermans, S.B. Gordon Clinical Microbiology and Infection Volume 20, Issue 12, Pages O1145-O1151 (December 2014) DOI: 10.1111/1469-0691.12752 Copyright © 2014 European Society of Clinical Infectious Diseases Terms and Conditions

FIG. 1 Proportion of carriage-positive volunteers detected by bacterial culture or quantitative PCR (qPCR) over time. For detection by culture, nasal wash samples were plated on blood agar with gentamicin. For qPCR detection, nasal wash was added to RNAprotect and frozen until DNA extraction. A volunteer was considered to be positive by culture (white bar) if Gram-positive, α-haemolytic, optochin-sensitive colonies were detected after 48 h. Serotype was confirmed by latex agglutination. The target gene for qPCR was lytA. Samples with a Ct value of <40 were considered to be qPCR-positive (black bar). The number of samples analysed is indicated above each time-point. *Statistical significance (p ≤0.05) Clinical Microbiology and Infection 2014 20, O1145-O1151DOI: (10.1111/1469-0691.12752) Copyright © 2014 European Society of Clinical Infectious Diseases Terms and Conditions

FIG. 2 Correlation between bacterial culture and qPCR in quantifying Streptococcus pneumoniae in nasal wash (NW) samples. Quantification of pneumococci by culture and by qPCR are positively correlated. The Spearman rank correlation coefficient for samples positive by both culture and qPCR is 0.73. Dotted lines represent lower limits of detection. A p-value of ≤0.05 was considered to be significant. Clinical Microbiology and Infection 2014 20, O1145-O1151DOI: (10.1111/1469-0691.12752) Copyright © 2014 European Society of Clinical Infectious Diseases Terms and Conditions

FIG. 3 Culture and quantitative PCR (qPCR) are complementary for following a carriage episode. (a, b) At low density, carriage results can vary by method, which can impact on (a) the number of carriage episodes and (b) the duration of carriage episodes. (c) Natural carriage episodes mimic the variation seen in experimental carriage. Filled shapes: carriage-positive by culture. Open shapes: carriage-negative by culture. Dotted lines represent lower limits of detection. NW, nasal wash. Clinical Microbiology and Infection 2014 20, O1145-O1151DOI: (10.1111/1469-0691.12752) Copyright © 2014 European Society of Clinical Infectious Diseases Terms and Conditions