We report the first laboratory-confirmed human infection with O'nyong-nyong virus in Chad. This virus was isolated from peripheral blood mononuclear cells of a patient with evidence of a seroconversion to a virus related to Chikungunya virus. Genome sequence was partly determined, and phylogenetic studies were conducted. O n November 2, 2004, a febrile 19-year-old French soldier staying in Chad and returning from a mission in Sarh, in the southern part of the country, was admitted to the hospital. Clinical examination showed a high body temperature (38°C), rash, periocular erythema, and pharyngitis. Abdominal, cardiopulmonary, and neurologic functions were normal. Except for body temperature, biochemical and hematologic values were normal. Serologic results were negative for Rickettsia typhi, R. conorii, Legionella pneumophila, Bordetella pertussis, HIV, and human herpesviruses 1 and 2. Results of malaria testing were also negative. The patient received intravenous acetaminophen for 2 days, according to the protocol used by the French Armed Forces Medical Service in the event of fever occurring overseas. He recovered after 5 days without sequelae.Serum samples, collected during the acute phase (November 3-5, 2004) and after (November 23 and December 7, 2004; January 10 and February 1, 2005) were transported to our laboratory and tested by ELISA for immunoglobulin M (IgM) and IgG antibodies to a battery of arboviruses by IgM-antibody capture (MAC-ELISA) and antigen-capture ELISA, respectively (1). Each serum sample was considered positive if the optical density (OD) ratio, OD (viral antigen)/OD (uninfected cells), was >3. The first sample (November 3, 2004) contained no antibodies (OD ratio <2) to dengue viruses, West Nile virus, Wesselsbron virus, Rift Valley fever virus, Bunyamwera virus, or Chikungunya virus (CHIKV). Remaining samples contained antibodies to a virus serologically related to CHIKV (OD ratios >3) for both IgM (sample 2 and following samples) and IgG (sample 3 and those following). Antibody titers peaked 20 days (IgM) and 68 days (IgG) after the onset of symptoms (Figure 1). The IgM titer returned to a low level within 2 months after onset of illness.Results of CHIKV-specific real-time reverse transcription-PCRs (2) performed with serum samples as templates were negative. Virus isolation was attempted by incubation of peripheral blood mononuclear cells collected on the day of onset with C6/36 (Aedes albopictus) and Vero (E6 clone) monolayers. After 5 days, supernatants were collected and used to infect fresh cell cultures. After 2 days, cytopathologic effects were observed in Vero monolayers; a high level of cell death was also observed in C6/36 cells. Infected Vero and C6/36 cells were then examined by indirect immunofluorescence assay (IFA) for 8 different alphaviruses with alphavirus-specific antibodies and inhouse mouse hyperimmune ascitic fluids to CHIKV, Mayaro (MAYV), Tonate (TONV), Semliki Forest (SFV), and Sindbis (SINV) viruses. Results of IFA were positive when alphavirus-specific a...