Dengue fever is a frequent cause of morbidity in travellers. The objective was to describe the epidemiological and clinical characteristics of dengue fever in Czech travellers. This descriptive study includes patients with acute dengue fever diagnosed at Hospital Na Bulovce during 2004-2013. Data were collected and analysed retrospectively. A total of 132 patients (83 males and 49 females) of median age 33 years (IQR 29-40) were included. Diagnosis was established by NS1 antigen detection in 87/107 cases (81.3%) and/or RT-PCR in 50/72 (69.4%) and by serology in 25 cases (18.9%). Dengue was acquired in South-East Asia in 69 cases (52.3%), followed by South Asia (48 cases; 36.3%), Latin America (14; 10.6%) and Sub-Saharan Africa (1; 0.8%). The most frequent symptoms included fever, rash and headache. Initial leukocyte and lymphocyte counts were lower in patients who presented in the early phase (0-4 days), however, platelet count was lower and AST, ALT and LDH activity higher in patients with a longer symptoms duration (≥5 days). The clinical course was mostly uncomplicated. Dengue fever is becoming a frequent cause of fever in Czech travellers. Clinicians should be familiar with the typical clinical findings and novel diagnostic methods.