Centered on molecular analysis, the present research aims to detect the occurrence of human papillomavirus (HPV16) 16 from cervical samples in Dhi-Qar Province, Iraq. From 2017 to 2020, this survey was conducted on 93 adult females with cervical cancer and 60 stable individuals as a control group. The patients' ages ranged from 32 to 78. For the molecular analysis, DNA was isolated and subjected to polymerase chain reaction (PCR) for amplification and identification of the minor capsid protein L2 gene. PCR results showed 60 (65%) from 93 cervical cancer cases infected by HPV16 while only 5 (8%) of the healthy control group are positive for HPV16. The present survey detected the most high-risk HPV16 infections in 2019 (78%) and 2020 (69%) in compared with the lowest infections (47%) in 2017. Also, the incidence of HPV16 infection varies by age group, as the current findings indicate that the majority of infections occur in young women, while infection rates decline in older women. On the other hand, the distribution of HPV16 infections according to cervical cancer stages revealed that stage IV (70 percent) had the most infections, followed by stage III (68 percent), and stage II (32 percent) (60 percent). PCR is a professional method for the precise detection of HPV16. Moreover, HPV16 infections significantly associated with cervical carcinoma among women in Dhi-Qar Province /Iraq, and viral infection increased during recent years, especially in young women, and clearly related to cervical cancer progression.