Dengue is an acute systemic viral disease that has been developed globally in both chronic and epidemic transmission periods. Dengue virus (DENV) is a member of the Flavivirus genus of the Flaviviridae family, which endangers public health. Limited studies have been performed in the Saudi Arabia and there are no epidemiological as well as molecular screening of DENV in the Southwestern region and this current study was conducted on the epidemiology of dengue in the Southwestern region of Saudi Arabia. Simultaneously, we have screened the 100 patients for DENV using the real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay. The current study results confirmed that 6637 people from various hospitals and locations in Jazan, Southwestern regions, were enrolled in this study from 2012 to 2020. The overall mean age was 30.02 ± 18.01 years, with 62.8% of males and 37.2% of females enrolled. This study included nearly three-fourths of the Saudi participants and one-fourth of the expatriates, and 56.6% of the positive cases were enrolled. In 2019, the most instances were enrolled, with 44% of positive cases. When screened using the RT-PCR assay, 93% of the positive patients were recruited, according to the quality control analysis. In conclusion, the current study results confirmed the prevalence of DENV was increased drastically since 2012 to 2020. High number of cases were registered prior to the Pandemic. The screening for DENV was performed with RT-PCR assay and NSI antigen should also be implemented to cross-check the results which was previously performed with RT-PCR analysis.