We studied the incidence of invasive pneumococcal disease (IPD) in the Czech Republic by analysing two sources of data. The incidence of pneumococcal meningitis based on routine notification data varied between 0·4 and 0·6/100 000 population between 1997 and 2006. The incidence of IPD based on laboratory surveillance varied between 2·3 and 4·3/100 000 population between 2000 and 2006. The annual IPD incidence remained stable during the study period. Estimates of absolute IPD case-load in the entire country varied from 235 to 437 per year. The age-specific incidence was highest in the <1 year age group, reaching 4·3/100 000 for pneumococcal meningitis in routine notification and 15·7/100 000 for IPD in laboratory-based surveillance data, respectively. A total of 1236 Streptococcus pneumoniae isolates from cerebrospinal fluid and sterile body sites were investigated. The most frequent serotypes causing IPD in all ages were 3, 4, 14, 8 and 19F, accounting for 41·5% of all isolates. The most frequent serotypes by age group were: <1 year (6B and 19F); 1–4 years (14, 6B and 23F); 40–64 years (3, 8 and 4), and ⩾65 years (3, 4, 9N and 14). The coverage of serotypes in all age groups by pneumococcal vaccines ranged from 41·5% for 7-valent conjugate vaccine to 67·9% for 13-valent conjugate vaccine. The coverage of serotypes causing IPD is significantly different between infants/children and adults/elderly. PCV-7 coverage by age group was: <1 year (66·0%), 1–4 years (65·1%), 40–64 years (34·4%) and ⩾65 years (39·3%). Similar age differences between infants/children and adults/elderly were found in coverage by PCV-9, PCV-11 and PCV-13. The distribution of serotypes in the total population and individual age groups was stable during the period 2000–2006.