Dengue fever represents an increasing arboviral public health concern in a significant endemic population situated in tropical and subtropical regions globally, exhibiting different levels of clinical manifestation. This research focused on examining the clinical and laboratory dynamics associated with the four dengue serotypes. This cross-sectional study was conducted in the Department of Microbiology Laboratory, Adesh Institute of Medical Sciences and Research, Adesh University, Bathinda, Punjab, India. Blood samples from all clinically dengue suspected patients were received from various Department of AIMSR hospital, between the period of August 2024 to July 2025. Samples were tested for NS1Ag, IgM and IgG by ELISA method. Dengue serotypes were detected by using RT-PCR at BSL-III laboratory of AIMSR hospital. During the study period, 378 samples were tested among which 39 were positive for one or more dengue parameters by ELISA method. 15 (38.46%) samples were positive for NS1Ag and 3 (7.69%) were positive for IgM only. Prevalence of Dengue was found to be (39) 10.30%. Among the 39 ELISA positive samples, prevalence of dengue serotypes amplified by RT-PCR. DENV-2 (51.28%) was most predominant serotypes. Other serotypes were significantly less common, with DENV-1 and DENV-3 identified in 7.69% each, and DENV-4 in 2.56%. 10.25% of cases showed co-infection with DENV-2 + DENV-3 serotypes, whereas 5.12% of cases exhibited co-infection with DENV-1 + DENV-2 and DENV-1 + DENV-3 serotypes each, DENV-3 + DENV-4 and all four-serotype identified in a single case accounting for 2.56% each. Most of the dengue patients in our study were residents of Punjab. Our study revealed that DENV-2 is the most common serotype with maximum severity. Dengue was more frequently diagnosed in females than males.