Background: Dengue is one of the most serious mosquito borne viral infection mainly affecting tropical and subtropical countries of the world. In absence of specific treatment and vaccine for dengue fever (DF); vector control is the only method by which the spread of dengue can be prevented. Aims & Objective: This study was conducted to report the prevalence of Dengue virus infection in Gujarat, Western India. Material and Methods: Study was performed at a tertiary care hospital in Ahmedabad, Gujarat in year 2012. Patients attending various hospitals across Gujarat for suspected dengue were tested. Blood samples collected in plain tubes were tested for dengue IgM antibodies, NS1 antigen and viral nucleic acid detection by Dengue IgM capture enzyme linked immune sorbent assay, Dengue Early ELISA and real time reverse transcriptase PCR, respectively. The laboratory records were analyzed for demographic features and seasonal variations. Descriptive statistics were used. Data were expressed in proportions. Results: Out of total 4401 serum samples tested, 927 were found positive for dengue virus infection. 65% positive samples were of male patients and 58% positive samples were from 18 to 35 years age group (Adult population) (58%). Seasonal trend showed a gradual increase in dengue positives started from August with a peak in October (34.5%). The most common presentation was fever (97%) while only 1% cases presented haemorrhagic manifestations. Conclusion: Dengue has established its transmission in urban and semi-urban areas of Gujarat with predominantly affecting males and active adult population. Virus activity is high during monsoon and post monsoon period which coincides with increased vector breeding. This study thus emphasizes the need for continuous sero epidemiological surveillance for the timely formulation and implementation of effective dengue control programme.