To assess the prevalence of HPV infection among women with cervical cancer in Tiruchirapalli, Tamil Nadu, India.A case–control study was conducted with 246 women with cervical cancer and 257 control participants aged between 20 and 70years. The presence of HPV DNA was determined using the MY09/11 PCR protocol, the GP5 + /6 + PCR protocol, and type-specific PCR-based assays.The overall HPV prevalence was 54.9% and HPV 16 was the most prevalent type. The women the most vulnerable for HPV infection were those aged 41 to 60years. The major contributing risk factors for cervical cancer were having had 3 or more pregnancies (OR 19.2; 95% CI, 10.78–10.16); harboring high-risk HPV DNA (OR 15.3; 95% CI, 9.81–23.8); being a manual worker (OR 14.9; 95% CI, 9.62–23.08); being illiterate (OR 8.28; 95% CI, 5.51–12.43); and having never been screened for cervical dysplasia (OR 6.70; 95% CI, 10.78–34.44).The present report on the overall and type-specific prevalence rates and risk burden of HPV infection in Tamil Nadu confirms that screening and vaccination programs are urgently needed in this state to overcome the burden of HPV-associated cervical cancer.