Virological investigation was carried out to determine the etiology of suspected Chikungunya fever among humans reported in the Lakshadweep islands in the Indian Ocean. Three out of 23 acute sera samples showed cytopathological changes in Vero cell lines. Further, indirect immunofluorescence antibody test and reverse transcriptase-polymerase chain reaction studies demonstrated the Chikungunya virus etiology during the episode. E1 gene sequence analysis has confirmed the involvement of the Central/East African genotype of the Chikungunya virus.