Background & Objectives: Ureaplasma infection in asymptomatic women shows a significance of women’s reproductive health. Culture-based methods are slow, labor-intensive, and can miss non-culturable or fastidious strains. Targeting 16SrRNA gene, a highly conserved genetic marker found in the bacterial genome, has been widely utilized for microbial identification and phylogenetic analysis. The main aim of this study is to identify the prevalent genotypes of Ureaplasma species by the application of specific 16SrRNA target in asymptomatic women. Methodology: This cross-sectional study was performed with 100 asymptomatic women aged from 18 to 45 years with infertility. Conventional PCR for 16SrRNA was taken to determine the presence of U. urealyticum and U. parvum using endocervical samples. Amplified PCR products of were analyzed and positive products were sent for sequencing and the results were compared with GenBank and phylogenetic analysis was constructed. Results & Discussion: The overall result was found that 24.0% of asymptomatic women were positive for Ureaplasma species. One of our study sequences were closely clustered with common ancestors of reference strains from Austria, Ireland, Canada and China. Remaining five sequences forms a separate ancestor lineage for the USA strain JQ901478.1. Conclusion: The study concludes that the major identification of Ureaplasma species in asymptomatic women is Ureaplasma parvum which represents a multifaceted challenge with significant clinical implications.