To determine the role of Mycoplasma pneumoniae (M. pneumoniae) in pediatric lower respiratory tract infections (LRTIs) employing serological tests and polymerase chain reaction (PCR) analysis. In this prospective study, 200 children aged 6 mo to 12 y hospitalized with acute LRTIs were investigated for M. pneumoniae. Serum samples were collected for serological analysis of M. pneumoniae. Throat swab samples were obtained on admission to amplify 277-base pair region of 16S rDNA gene of M. pneumoniae by PCR. In the present study, 40(26.1%) children <5 y and 28(59.5%) children ≥5 y age group were positive for M. pneumoniae infection and this difference was statistically significant (P < 0.001). M. pneumoniae was positive in 32(41%) female and 36(29.5%) male children though this difference was statistically insignificant (P = 0.12). The clinical profile across M. pneumoniae positive and negative cases were comparable except for presence of chest pain which was statistically significant (P = 0.023). None of the radiological findings was statistically associated with incidence of M. pneumoniae infection. Serological evidence of acute M. pneumoniae infection was observed in 64(32%) patients with sensitivity 66.6% and specificity 70.1% while PCR positivity in 12(6%) patients with sensitivity 12.5% and specificity 97%. Together, serology and PCR detected M.pneumoniae infection in 68(34%) patients. The present study underlines the role of M. pneumoniae in children with community- acquired LRTIs and more particularly in ≥5 y of age.