Wild indigenous small mammals including 83 rodents (bandicoot and black rats, and house mice) and a shrew captured from multiple sites in Vellore, south India, were tested for serological and molecular evidence of hantavirus infection. Indirect immunofluorescence assay (IFA) using Hantaan virus (HTNV) antigen indicated hantavirus-reactive antibodies in 16 (19.3%) of 83 rodents (bandicoot and black rats). Western blot (WB) using Thailand virus (THAIV) antigen confirmed hantavirus-reactive antibodies in nine of the 16 HTNV IFA-positive rodents. Reverse transcription polymerase chain reaction (RT-PCR) of lung and kidney tissue of captured mammals resulted in the detection of partial S segment sequence in a bandicoot rat. This study complements our earlier reports on hantavirus epidemiology in south India and documents first laboratory evidence for rodent-associated hantaviruses in south India.