Genomic Identification of Genital Ureaplasma urealyticum and Ureaplasma parvum Infection by Duplex Polymerase Chain Reaction in Symptomatic Women Attending a Tertiary Care Hospital: A Cross-sectional Study

Abstract

Introduction: Ureaplasma urealyticum (U. urealyticum) and Ureaplasma parvum (U. parvum) are the two causative agents for Sexually Transmittable Diseases (STDs), they are often accompanied by Pelvic Inflammatory Disease (PID), vaginitis, endometriosis, etc. Aim: To detect the prevalent genotypes/biovars of Ureaplasma species by using Duplex Polymerase Chain Reaction (PCR) in symptomatic women attending the Obstetrics and Gynaecology clinic of a tertiary care hospital. Materials and Methods: This was a cross-sectional study, comprised of 200 symptomatic women aged from 18 to 45 years. This study was carried out in the Department of Microbiology, School of Allied Health Sciences, Department of Obstetrics and Gynaecology, Mahatma Gandhi Medical College and Research Institute, and Mahatma Gandhi Medical Advanced Research Institute, Sri Balaji Vidyapeeth, Puducherry, India for the period of January 2021 to January 2024. Duplex PCR was performed for the presence of U. urealyticum and U. parvum in endocervical samples. Amplified PCR products were analysed and the positive PCR products were sent for sequencing and the sequenced results were compared with GenBank and phylogenetic analysis was constructed. Data was analysed using Chi-square tests to test level of significance between the two groups (p-value≤0.05). Results: The overall results found that 48 (24%) of symptomatic women were positive for Ureaplasma species. In Duplex PCR, 24 (12.0%) were positive for both urease and upv genes; followed by 15 (7.5%) for urease and 9 (4.5%) for upv genes only. The phylogenetic tree concluded that the genetic sequences of Ureaplasma species are closely related to various isolates within a species. Conclusion: The study concludes that U. parvum is the most common pathogen in symptomatic women with predominant age of 26-35-year-old. The duplex PCR, increase the sensitivity and specificity by the application of two different genes in a single amplification reaction. Since the research will focus on different gene targets of Ureaplasma species by the application of multiplex in future study.

Keywords

• pelvic inflammatory disease
• sexually transmitted diseases
• urease gene
• upv gene
• vaginal discharge