Background: Genital infections caused by Herpes Simplex viruses are viewed as the indicators of sexual network in the society. Most of the infections caused by these viruses are asymptomatic and underdiagnosed. Aim: To investigate the seroprevalences of HSV-1 and HSV-2 among the STI vulnerable women and correlate them with clinical infections and other co-viral STIs. Study design: A cross sectional retrospective study was conducted on female contacts of HIV / STI positive male partners, who were consulting the STD clinics of NGOs and OP section of STD department of Government hospital in Chennai, India between October 2006 and September 2008. Methodology: The study included 138 STI vulnerable women along with age matched control group from whom socio-demographic and clinical data were collected. Serological screening was performed using ELISA for detection of IgM and IgG antibodies to HSV types 1 and 2 and co-positivities to HIV, HBV, HCV and CMV. Statistical analysis of results was carried out using Chi-square test. Results: Overall seroprevalences of 79.71% and 74.32% were recorded respectively for HSV-1 and HSV-2 among the study group. Proportionate increase in symptomatic cases were observed with individuals showing seropositivities to anti-HSV-1/2 IgM, IgM+IgG (combined) and IgG antibodies, which correlated respectively with primary, reactivated and episodic infections. Observation of symptomatic cases among HSV-1 (48.0% vs. 4.05%) and HSV-2 (68.29% vs. 25.0%) IgG positive study and control groups corroborated the sexual transmission of these viruses. Significant percentages of co-positivities to other viral STIs were recorded with higher preponderance among HSV-2 seropositive individuals than those of HSV-1. Conclusion: Comprehensive serological screening and its correlation with the clinical data would be very helpful in demonstrating the impact of HSV infections. Wide screening of STI vulnerable women is necessary to estimate the actual burden of STIs and to adopt preventive intervention.