Introduction: It is estimated that almost one-quarter of worldwide malignancy can be associated with infectious agents, of which approximately one third is caused by HPV. Cervical cancer caused by persistent infection with high-risk HPV genotypes is the second leading cause of death in women aged 15 to 44. Tests based on molecular biological methods commonly used today in diagnosing HPV infections enable genome detection and determination of individual HPV genotypes in cervical smear samples. However, this diagnostic approach does not provide insight into the type of infection and assessment of the risk of cervical cancer. In recent years, numerous studies have focused on improving the primary test to find an appropriate diagnostic method to provide insight into the type of infection. The study’s objectives were to determine the oncogenic activity of the most commonly diagnosed high-risk HPVs in cervical smear samples using the mRNA test and compare the results according to the severity of the cervical intraepithelial lesion. Material and methods: The study included 365 samples of cervical swabs of women older than 18 years, referred by gynecologists to the Center for Virology, Institute of Public Health of Vojvodina from 2017 to 2021. According to the Bethesda classification, samples were grouped according to the severity of the cervical intraepithelial lesion (NILM, ASCUS, LSIL, HSIL). There were at least 25 samples of cervical smears in each group in which the presence of HPV DNA was proven. Demographic data and risk factors that contribute to the acquisition and persistence of HPV infection of the cervical epithelium were recorded. Using a qualitative real time PCR test, HPV DNA was detected and genotyped in cervical smear samples, while the reverse transcription and amplification method proved the presence of mRNA oncogenes E6 and E7, the oncogenic activity of tested HPV. Statistical analysis was performed using parametric and nonparametric tests, correlation, univariate, and multivariate logistic regression analysis. The results indicate that the most common genotypes in the female population of South Ba��ka District are HPV 16, 31, 33, and 51. In 84.3% of the analysed samples, the infection caused by a single genotype was proven, while in 15.7% of cases, a co-infection is present. A statistically significant difference was found for HPV 16 (p = 0.035) and HPV 31 (p = 0.017) depending on the severity of the cytological results, which did not determine for HPV 33 and 51. The presence of HPV 16 is equally present in all age categories. The detection of HPV 31 and 33 genotypes decreases with years of life, while HPV 51 increases. The oncogenic activity was demonstrated in 64.5% of HPV-positive women. A statistically significant difference was found in the number of positive mRNA E6/E7 HPV 16 results concerning cytological status (p = 0.000) and age categories (p = 0.026). Approximately every other HPV 16 genotype was oncogenically active (48.6%). It was most prevalent in women with HSIL cytological results (64.2%), while the presence of mRNA E6/E7 HPV 31 was minimally detected in HSIL (7.5%). The distribution of oncogenic activity of the remaining genotypes is approximately the same through different cytological groups and remains at low level (2.2% ��� 11.4%). A comparison of tests for the detection of HPV and their oncogenic activity to assess the progression of cervical intraepithelial lesion indicated that statistically significantly higher specificity (89.1%) and positive predictive value (69.8% ��� 78.7%) were expressed for mRNA E6/E7 test, while significantly higher sensitivity is recorded when using the HPV DNA test (67.6% ��� 88%). The results of the ROC curve determine the higher probability of detecting HPV infection by 7% provided by the mRNA test. A statistically significant final model was defined. The oncogenic activity of HPV 16 and the age category of HPV-positive women were named as the strongest predictors for the development of precancerous lesions. Analysis of the parameters of the observed demographic factors and risk factors of HR HPV-positive subjects showed a difference concerning the severity of the cervical lesion. Conclusions: The largest number of infected women in the South Ba��ka District is caused by high-risk genotypes HPV 16, 31, 33, 51, which are oncogenic in more than half of the examined cases. The total oncogenic activity increases with increasing severity of the cervical intraepithelial lesion, with oncogenic active HPV 16 being most commonly detected in high-grade cervical lesions. In contrast, the prevalence of oncogenic active HPV 31 is minimally detected in high-grade lesions, while the prevalence of HR HPV 33 and 51 is low and not proportional to the severity of the cervical lesion. The prevalence of HPV infection is inversely proportional to the age of women in the South Ba��ka District, which indicates that regression of infection is the dominant process following the adequate immune response. The higher specificity and positive predictive value of the mRNA test than the HPV DNA test indicate a higher probability of detecting persistent HPV infection. The oncogenic activity of the most commonly detected high-risk HPVs has a predictive potential in assessing the occurrence of high-grade cervical epithelial lesions. The oncogenic activity of HPV 16 and age were the biomarkers with the strongest predictive values for the development of these lesions.