Introduction: Sexually transmitted Infections (STIs) are major public health concerns reaching an all-time high, globally. In Malaysia data on the prevalence of STIs remains scarce which limits the understanding of STI transmission dynamics and the role of interventions in the control of STIs. The aim of this study is to determine the epidemiology characteristics of STIs mainly from Malaysian private healthcare institutions. Method: A two years (2016 and 2017) retrospective review was conducted on 160 multiplex RT-PCR STI reports from KPJ hospitals, Malaysia. Results: There were 65 (40.6 %) patients positive STIs [male: 21/65 (32.3 %); female: 44/65 (67.7 %)]. The STIs was prevalent among young adults (56/65; 86.2 %) from the central region (46/65; 70.8 %). Females had 1.7 times greater risk to develop STI (20 per 100) and two times higher chance to have multiple STIs (10 per 100) in comparison to male. The single STIs was caused mainly by U. parvum (N=17). In males, U. urealyticum (N=3) and C. trachomatis (N=3) were prevalent, while U. parvum (N=15) was prevalent in females. There were 19 dual infections of STIs which were commonly caused by U. parvum and M. hominis (N=5). There were seven STIs cases caused by three pathogens concurrently including U. urealyticum, U. parvum and C. trachomatis (N=2) and U. urealyticum, M. genitalium and C. trachomatis (N=2). Females from urban communities have higher risk in comparison to males for developing multiple STIs. Conclusion: This study provides an imperative platform for temporal trends of STIs in Malaysia which reflects the health status of certain populations that warrant immediate public health interventions.