Mycoplasma pneumoniae has been well established as a major cause of lower respiratory tract infections (LRTIs) accounting for 10-30% of all cases of community-acquired pneumonia and is clinically indistinguishable from other infectious causes of pneumonia. The study aimed to investigate the occurrence of M. pneumoniae as the etiologic pathogen in LRTIs among children and to compare Polymerase Chain Reaction [PCR] and serology for the diagnosis of M. pneumoniae in community-acquired LRTIs in children. A total of 133 children aged between 6 months and 12 years with signs and symptoms of community-acquired LRTIs attending the Paediatrics OPD, emergency or admitted to the wards of a tertiary care hospital were prospectively enrolled into the study. M. pneumoniae in throat swab samples was detected by conventional PCR, and compared with serology and clinical signs and symptoms. Univariate analyses was conducted to determine the association of M. pneumoniae infection among different categories of patients. 31 out of 133 patients included in the study (23.3%) were positive for M. pneumoniae by any test. Among 31 patients, serology (IgM) was positive in 19 patients (61.2%) and PCR in 12 patients (38.7%); 2 patients were found to be positive by both methods. Fever, cough, hurried breathing and intercostal retraction were the clinical signs and symptoms significantly associated with LRTIs due to M. pneumoniae (P<0.05). A combination of clinical features, PCR and IgM for Mycoplasma pneumoniae is recommended depending upon the duration of illness for optimal diagnosis, timely initiation of therapy and to prevent overuse of macrolides.