Detection of human papillomavirus (HPV) genotypes by pyrosequencing method in uterine cervical cancer samples

Cervical cancer is the second most common type of cancer in women after breast cancer. Human papillomavirus (HPV) is considered as the primary etiologic agent of cervical cancer. HPV types is categorized according to the epidemiological relationships and genotypes of HPVs. According to analysis of genome sequences of HPVs, HPV genotypes more than 100 have been identified. The molecular tests using for detection of HPV genotypes are very important for the detection of vaccine strategy and epidemiological purpose. The pyrosequencing method is one of the new method using for HPV genotypes. In this thesis, detection of HPV genotypes were aimed by pyrosequencing method in patients with uterine cervical cancer. For this study, 38 paraffin-embedded cervical cancer samples obtained from Department of Pathology, Faculty of Medicine, Firat University were used. The presence HPV DNA in samples were determined by real time-PCR. Then, HPV genotypes were determined from HPV DNA positive samples using pyrosequencing method. According to the results of real time-PCR, 35 (92%) of 38 samples were found as positive for HPV DNA. In the result of pyrosequencing method, of 35 HPV DNA positive samples HPV 16 in 26 (68.42%), HPV 18 in 2 (5.26%) were detected. In one sample, HPV 16 and HPV 18 super-infection (2.63%). In other one sample, HPV 16 and HPV 33 super-infection (2.63%) were detected. HPV 16 ve Africian subtip süper enfeksiyonu (%13.15) were detected fron in 5 samples. According to these results, HPV 16 and HPV 18 were shown to be the most important genotypes. Identification of HPV genotypes with pyrosequencing is cheaper and easier as compared to other methods such as classic nucleotite sequensing method. Thus, we suggest to use pyrosequencing method for genotyping in routine laboratories. Key words: Human papilloma virus, HPV, uterine cervical cancer, genotype, pyrosequencing.