Human Papilloma virus infection is the fundamental reason for the development of ano-genital malignancies and knowing the best tool for diagnostic purposes is mandatory. This study investigated the prevalence and genotype distribution of HPV genotypes in formalin-fixed paraffin-embedded (FFPE) blocks from patients with different anogenital lesions. In this cross-sectional retrospective study, 125 blocks from patients with different anogenital lesions were collected. Three internal sections were taken for HPV detection and genotyping using the paraffin tissue processing kit and HPV Direct Flow CHIP. HPV positivity was detected in 90 (72.0%), with 77 (85.6%) females and 13 (14.4%) males as follows: SCC 64.0%, CINIII 66.7%, CINII 100.0%, CINI 83.3%, KA 83.7%, NILM 44.0%, Anus 66.6%. A total of 44% of histologically unremarkable (negative) cases were positive for HPV genotypes while in only 64% of SCC were HPV genotypes detected. Sixty-six (73.3%) cases were low-risk, and 16 (17.8%) cases were high-risk genotypes, mostly cervical lesions, while seven (7.8%) cases showed a mixed viral detection. The most frequent low-risk genotype was HPV6 genotype (62–68.9%), while the prevalent high-risk HPV was HPV16 genotype (12–13.3%). In this study, HPV16 was more frequently detected than HR-HPV, but mainly in cervical lesion, while HPV6 topped the LR-HPV infections amongst different anogenital lesions in Duhok-Iraq. Higher HPV positivity among cytological unremarkable and abnormal cases may reflect the higher sensitivity of the direct flow CHIP diagnostic technique. The results demonstrate that screening for HPV is essential to reduce cancer development and planning for the vaccine’s introduction in Iraq.