Density and duration of pneumococcal carriage are considered to affect the likelihood of transmission and invasive disease. Because of its importance in both spreading and causing disease, carriage has been suggested as an endpoint in future vaccine studies. Culture is the current gold standard for detection but may not be sensitive enough to detect changes at low density. Healthy adult volunteers received an intranasal inoculation of Streptococcus pneumoniae serotype 6B. Pneumococcal density in nasal washes collected at six time points post-inoculation was determined by culture and qPCR. Natural pneumococcal carriers detected at initial screening were followed in parallel. In 331 nasal washes from 79 volunteers, the sensitivity and specificity of pneumococcal detection by qPCR, compared with culture, were 92.3% and 75.9%. The estimation of pneumococcal density by culture and qPCR was highly correlated (rs=0.73, P<0.0001), although qPCR had a lower detection limit. Pneumococcal density fluctuated within a carriage episode and occasionally fell under the detection limit of both methods. The duration of carriage episodes was underestimated when using only one method. Similar fluctuations in density were observed in natural carriers. Pneumococcal carriage is a dynamic event. Culture and qPCR are complementary in surveying density and duration of pneumococcal carriage episodes.This article is protected by copyright. All rights reserved.