Molecular diagnosis of genital mycoplasma and ureaplasma species by Duplex PCR and real time PCR in symptomatic and asymptomatic women attending a tertiary care hospital in Puducherry

Abstract

newline ABSTRACT newlineBackground newlineImportance of sexually transmitted infections, such as Ureaplasma and Mycoplasma newlinespecies, in reproductive health issues is becoming more widely acknowledged. These bacteria avoid newlineimmune surveillance by avoiding detection because to their tiny genome and absence of a cell wall, newlinewhich can lead to persistent infections that affect fertility. The present study aims to determine the newlineprevalence and genotypes of Ureaplasma and Mycoplasma species in symptomatic and newlineasymptomatic women in Puducherry, India. newlineObjectives newlineThe main objectives include detecting Ureaplasma and Mycoplasma genes in endocervical newlinesamples using Duplex PCR and Real-Time PCR, followed by gene sequencing for molecular newlinecharacterization. newlineMethods newlineA cross-sectional study was conducted among 410 women (205 symptomatic and 205 newlineasymptomatic) from September 2020 to September 2024. Endocervical swabs were collected, and newlinegenomic DNA was extracted. Duplex PCR and Real-Time PCR were performed to identify specific newlinegenes of Ureaplasma (U. parvum, U. urealyticum) and Mycoplasma (M. hominis, M. genitalium). newlineGene sequencing and phylogenetic analysis were performed for molecular characterization. newlineStatistical analysis was conducted using the chi-square test and Fisher s exact test. newlineResults newlineAmong symptomatic women, 109 (53.17%) tested positive for either Ureaplasma or newlineMycoplasma, whereas 31 (15.12%) of asymptomatic women were also positive. Ureaplasma was newlinemore prevalent 73 (35.6%) than Mycoplasma 36 (17.5%) in symptomatic individuals. Duplex PCR newlineand Real-Time PCR demonstrated reliable sensitivity, with gene sequencing confirming genetic newlinediversity among isolates. newlineConclusion newlineThe study concludes that the Duplex PCR and Real-Time PCR is a valuable tool for newlinedetecting Ureaplasma and Mycoplasma species, in symptomatic as well as asymptomatic cases with newlinelow bacterial loads. The study also found evidence of co-infection with various bacterial, fungal and newlineparasitic diseases, including Bacterial vaginosis, Candida albicans and T. vaginalis.