Background Persistent infection with high-risk (HR) human papillomavirus (HPV) types is necessary for cervical cancer development. The risk is much higher for women infected with multiple genotypes of the human papillomavirus (HPV). The aim of this study was to evaluate the frequency of single and multiple HPV infection in relation to age and results of Pap smear cytology.

Methods From January to December 2012, frequency of HR HPV was analysed among 398 women from Vojvodina. In determination of the presence HPV DNA commercial HR HPV Real-TM kit (Sacace Biotechnologies, Italy) were applied. PCR was performed and products were detected using ABI 7500 Real-Time PCR instrument.

Results The overall prevalence of HPV infection among studied women was 61.3%. Single HPV genotypes were found in 150 (65.3%) and multiple genotypes in 94 (34.7%) of 244 HPV DNA positive samples. The most frequent genotype was HPV 16, followed by HPV 31, 51.52 and 18. Proportion of multiple HPV infection decreased significantly according to age and severity from 48.9% in women under 30 years of age, 39.7% in women from 30 to 40 years of age, 21.3% in women older than 40. Among all women with normal cytology multiple HPV infections were found in (22.4%), (34.9%) among women with ASCUS cytology, (17.0%) in LSIL cytology and (17.9%) in HSIL cytology.

Conclusion In the present study, we observed a very high prevalence of multiple (22.4%) of HPV infection among women with normal cytology and (34.9%) in women with indeterminate Pap test, as ASCUS cytology. Sensitive and specific detection methods, as well as, real-time PCR genotyping HPV are required and helpful for triage of women with ASCUS cytology.